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49 results on '"Biologie de la Reproduction, Environnement, Epigénétique '

1. BPA disrupts meiosis I in oogonia by acting on pathways including cell cycle regulation, meiosis initiation and spindle assembly

Author

Benoit Loup, Elodie Poumerol, Luc Jouneau, Paul A. Fowler, Corinne Cotinot, Béatrice Mandon-Pépin, Biologie de la Reproduction, Environnement, Epigénétique & Développement (BREED), École nationale vétérinaire - Alfort (ENVA)-Université de Versailles Saint-Quentin-en-Yvelines (UVSQ)-Université Paris-Saclay-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), University of Aberdeen, European Project: 212885,EC:FP7:ENV,FP7-ENV-2007-1,REEF(2008), and European Project: 080388,WT::(2006)

Subjects
cumulus-oocyte complex, Meiosis I, Stimulated by Retinoic acid gene 8, COC, Toxicology, spindle assembly checkpoint, Organ culture, retinoic acid receptor agonist, Oogonia, Fetal ovary Organ culture Meiosis Oogonia BPA Sheep RA, Bisphenol A, Phenols, Retinoic acid, Animals, Humans, STRA8, Benzhydryl Compounds, [SDV.BDD]Life Sciences [q-bio]/Development Biology, Sheep, Fetal ovary, days post coitum, PGC, SAC, DPC, BPA, Meiosis, primordial germ cell, Oocytes, Female, AM580
Abstract

International audience; The negative in utero effects of bisphenol A (BPA) on female reproduction are of concern since the ovarian reserve of primordial follicles is constituted during the fetal period. This time-window is difficult to access, particularly in humans. Animal models and explant culture systems are, therefore, vital tools for investigating EDC impacts on primordial germ cells (PGCs). Here, we investigated the effects of BPA on prophase I meiosis in the fetal sheep ovary. We established an in vitro model of early gametogenesis through retinoic acid (RA)-induced differentiation of sheep PGCs that progressed through meiosis. Using this system, we demonstrated that BPA (3 ×10-7 M & 3 ×10-5 M) exposure for 20 days disrupted meiotic initiation and completion in sheep oogonia and induced transcriptomic modifications of exposed explants. After exposure to the lowest concentrations of BPA (3 ×10-7 M), only 2 probes were significantly up-regulated corresponding to NR2F1 and TMEM167A transcripts. In contrast, after exposure to 3 × 10-5 M BPA, 446 probes were deregulated, 225 were down- and 221 were up-regulated following microarray analysis. Gene Ontology (GO) annotations of differentially expressed genes revealed that pathways mainly affected were involved in cell-cycle phase transition, meiosis and spindle assembly. Differences in key gene expression within each pathway were validated by qRT-PCR. This study provides a novel model for direct examination of the molecular pathways of environmental toxicants on early female gametogenesis and novel insights into the mechanisms by which BPA affects meiosis I. BPA exposure could thereby disrupt ovarian reserve formation by inhibiting meiotic progression of oocytes I and consequently by increasing atresia of primordial follicles containing defective oocytes.

Published
2022

2. Female age and parity in horses: how and why does it matter?

Author

Morgane Robles, E. Palmer, E. Derisoud, Pascale Chavatte-Palmer, Juliette Auclair-Ronzaud, Biologie de la Reproduction, Environnement, Epigénétique & Développement (BREED), Université Paris-Saclay-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-École nationale vétérinaire d'Alfort (ENVA)-Université de Versailles Saint-Quentin-en-Yvelines (UVSQ), Institut Français du Cheval et de L'équitation (IFCE), Académie d'Agriculture de France, and IFCE CS_2018_23

Subjects
puberty, endocrine system, placenta, 040301 veterinary sciences, animal diseases, embryo, Physiology, lactation, Reproductive technology, follicle, reproduction, 0403 veterinary science, Endocrinology, Pregnancy, mare, biology.animal, Genetics, Animals, Birth Weight, Horses, oocyte, Molecular Biology, fertility, uterus, biology, 0402 animal and dairy science, [SDV.BDLR]Life Sciences [q-bio]/Reproductive Biology, Embryo, 04 agricultural and veterinary sciences, 040201 dairy & animal science, Breed, Parity, Reproductive Medicine, Foal, Pregnancy, Animal, Gestation, Female, ovary, Animal Science and Zoology, Folliculogenesis, Parity (mathematics), Embryo quality, Maternal Age, foal, Developmental Biology, Biotechnology
Abstract

International audience; Although puberty can occur as early as 14-15 months of age, depending on breed and use, the reproductive career of mares may continue to advanced ages. Once mares are used as broodmares, they will usually produce foals once a year until they become unfertile, and their productivity can be enhanced and/or prolonged through embryo technologies. There is a general consensus that old mares are less fertile, but maternal age and parity are confounding factors because nulliparous mares are usually younger and older mares are multiparous in most studies. This review shows that age critically affects cyclicity, folliculogenesis, oocyte and embryo quality as well as presence of oviductal masses and uterine tract function. Maternal parity has a nonlinear effect. Primiparity has a major influence on placental and foal development, with smaller foals at the first gestation that remain smaller postnatally. After the first gestation, endometrial quality and uterine clearance capacities decline progressively with increasing parity and age, whilst placental and foal birthweight and milk production increase. These combined effects should be carefully balanced when breeding mares, in particular when choosing and caring for recipients and their foals.

Published
2021

3. Major transcriptomic, epigenetic and metabolic changes underlie the pluripotency continuum in rabbit preimplantation embryos

Author

Wilhelm Bouchereau, Luc Jouneau, Catherine Archilla, Irène Aksoy, Anais Moulin, Nathalie Daniel, Nathalie Peynot, Sophie Calderari, Thierry Joly, Murielle Godet, Yan Jaszczyszyn, Marine Pratlong, Dany Severac, Pierre Savatier, Véronique Duranthon, Marielle Afanassieff, Nathalie Beaujean, Institut cellule souche et cerveau / Stem Cell and Brain Research Institute (SBRI), Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Biologie de la Reproduction, Environnement, Epigénétique & Développement (BREED), École nationale vétérinaire - Alfort (ENVA)-Université de Versailles Saint-Quentin-en-Yvelines (UVSQ)-Université Paris-Saclay-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Interactions Cellules Environnement - UR (ICE), VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS), Isara, Cardiovasculaire, métabolisme, diabétologie et nutrition (CarMeN), Université de Lyon-Université de Lyon-Hospices Civils de Lyon (HCL)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Institut de Biologie Intégrative de la Cellule (I2BC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), Institut de Génomique Fonctionnelle - Montpellier GenomiX (IGF MGX), Institut de Génomique Fonctionnelle (IGF), Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université de Montpellier (UM)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université de Montpellier (UM)-BioCampus (BCM), Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université de Montpellier (UM)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université de Montpellier (UM), Unité sous contrat plate-forme biotechnologique de cellules souches plate-forme PrimaStem, Institut National de la Recherche Agronomique (INRA)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Institut National de la Santé et de la Recherche Médicale (INSERM), Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Institut cellule souche et cerveau / Stem Cell and Brain Research Institute (U1208 Inserm - UCBL1 / SBRI - USC 1361 INRAE), Institut méditerranéen d'océanologie (MIO), and Institut de Recherche pour le Développement (IRD)-Aix Marseille Université (AMU)-Institut national des sciences de l'Univers (INSU - CNRS)-Université de Toulon (UTLN)-Centre National de la Recherche Scientifique (CNRS)

Subjects
Pluripotent Stem Cells, Embryo transcriptome, Pluripotency continuum, Epigenesis, Genetic, Single-cell RNAseq, Mice, Blastocyst, Rabbit preimplantation embryo, Animals, Rabbits, Naive pluripotency, Transcriptome, Molecular Biology, [SDV.BDD]Life Sciences [q-bio]/Development Biology, Germ Layers, Developmental Biology
Abstract

Despite the growing interest in the rabbit model for developmental and stem cell biology, the characterization of embryos at the molecular level is still poorly documented. We conducted a transcriptome analysis of rabbit preimplantation embryos from E2.7 (morula stage) to E6.6 (early primitive streak stage) using bulk and single-cell RNA-sequencing. In parallel, we studied oxidative phosphorylation and glycolysis, and analysed active and repressive epigenetic modifications during blastocyst formation and expansion. We generated a transcriptomic, epigenetic and metabolic map of the pluripotency continuum in rabbit preimplantation embryos, and identified novel markers of naive pluripotency that might be instrumental for deriving naive pluripotent stem cell lines. Although the rabbit is evolutionarily closer to mice than to primates, we found that the transcriptome of rabbit epiblast cells shares common features with those of humans and non-human primates.

Published
2022

4. Dopaminergic and serotonergic changes in rabbit fetal brain upon repeated gestational exposure to diesel engine exhaust

Author

Karine Badonnel, Pascaline Bouillaud, Jacques Callebert, Henri Schroeder, Christine Baly, Benoit Olivier, Estefania Bernal-Meléndez, Pascale Chavatte-Palmer, Marie-Annick Persuy, Neurobiologie de l'olfaction (NBO), Institut National de la Recherche Agronomique (INRA), Composés Alimentaires : Biofonctionnalités et risques Neurotoxiques (CALBINOTOX), Université de Lorraine (UL), Service de biochimie et biologie moléculaire, Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Hôpital Lariboisière-Fernand-Widal [APHP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-IFR139, Biologie de la Reproduction, Environnement, Epigénétique & Développement (BREED), Université Paris-Saclay-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-École nationale vétérinaire d'Alfort (ENVA)-Université de Versailles Saint-Quentin-en-Yvelines (UVSQ), and ANR-13-CESA-0011,EPPAP,Effets de la Pollution Atmosphérique sur la fonction Placentaire et le développement Post-natal(2013)

Subjects
Male, 0301 basic medicine, Serotonin, Diesel exhaust, medicine.medical_specialty, Time Factors, Dopamine, Gestational exposure, Health, Toxicology and Mutagenesis, Hippocampus, 010501 environmental sciences, Biology, Toxicology, Serotonergic, Synaptic Transmission, 01 natural sciences, Norepinephrine, 03 medical and health sciences, Sex Factors, Pregnancy, Internal medicine, Monoaminergic, medicine, Animals, Vehicle Emissions, 0105 earth and related environmental sciences, Temporal cortex, Tyrosine hydroxylase, [SCCO.NEUR]Cognitive science/Neuroscience, [SDV.BA]Life Sciences [q-bio]/Animal biology, Dopaminergic, Neurotoxicity, Brain, Airborne pollution, General Medicine, Monoaminergic neuromodulation, Tryptophan hydroxylase, medicine.disease, 3. Good health, [SDV.BDD.EO]Life Sciences [q-bio]/Development Biology/Embryology and Organogenesis, 030104 developmental biology, Endocrinology, Maternal Exposure, Prenatal Exposure Delayed Effects, Female, Rabbits, [SDV.AEN]Life Sciences [q-bio]/Food and Nutrition
Abstract

International audience; Limited studies in humans and in animal models have investigated the neurotoxic risks related to a gestational exposure to diesel exhaust particles (DEP) on the embryonic brain, especially those regarding monoaminergic systems linked to neurocognitive disorders. We previously showed that exposure to DEP alters monoaminergic neurotransmission in fetal olfactory bulbs and modifies tissue morphology along with behavioral consequences at birth in a rabbit model. Given the anatomical and functional connections between olfactory and central brain structures, we further characterized their impacts in brain regions associated with monoaminergic neurotransmission. At gestational day 28 (GD28), fetal rabbit brains were collected from dams exposed by nose-only to either a clean air or filtered DEP for 2 h/day, 5 days/week, from GD3 to GD27. HPLC dosage and histochemical analyses of the main monoaminergic systems, i.e., dopamine (DA), noradrenaline (NA), and serotonin (5-HT) and their metabolites were conducted in microdissected fetal brain regions. DEP exposure increased the level of DA and decreased the dopaminergic metabolites ratios in the prefrontal cortex (PFC), together with sex-specific alterations in the hippocampus (Hp). In addition, HVA level was increased in the temporal cortex (TCx). Serotonin and 5-HIAA levels were decreased in the fetal Hp. However, DEP exposure did not significantly modify NA levels, tyrosine hydroxylase, tryptophan hydroxylase or AChE enzymatic activity in fetal brain. Exposure to DEP during fetal life results in dopaminergic and serotonergic changes in critical brain regions that might lead to detrimental potential short-term neural disturbances as precursors of long-term neurocognitive consequences.

Published
2021

5. Dynamics of cattle sperm sncRNAs during maturation, from testis to ejaculated sperm

Author

Hélène Kiefer, Andrea Rau, Eli Sellem, Sylvain Marthey, Chrystelle Le Danvic, Aurelie Bonnet, Laurent Schibler, Hélène Jammes, Luc Jouneau, Benoît Guyonnet, Biologie de la Reproduction, Environnement, Epigénétique & Développement (BREED), École nationale vétérinaire - Alfort (ENVA)-Université de Versailles Saint-Quentin-en-Yvelines (UVSQ)-Université Paris-Saclay-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Allice, Génétique Animale et Biologie Intégrative (GABI), AgroParisTech-Université Paris-Saclay-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Mathématiques et Informatique Appliquées du Génome à l'Environnement [Jouy-En-Josas] (MaIAGE), Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Transfrontalière BioEcoAgro - UMR 1158 (BioEcoAgro), Université d'Artois (UA)-Université de Liège-Université de Picardie Jules Verne (UPJV)-Université du Littoral Côte d'Opale (ULCO)-Université de Lille-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-JUNIA (JUNIA), Université catholique de Lille (UCL)-Université catholique de Lille (UCL), APIS-GENE (SeQuaMol), ANR-14-LAB3-0008,SeQuaMol,Qualité de la semence et fertilité chez les bovins: développement de nouveaux outils moléculaires de diagnostic(2014), École nationale vétérinaire d'Alfort (ENVA)-Université de Versailles Saint-Quentin-en-Yvelines (UVSQ)-Université Paris-Saclay-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Université Paris-Saclay-AgroParisTech-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Transfrontalière BioEcoAgro (Transfrontalière BioEcoAgro), and Université d'Artois (UA)-Université de Liège-Université de Picardie Jules Verne (UPJV)-Université du Littoral Côte d'Opale (ULCO)-Université de Lille-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)

Subjects
Male, endocrine system, Piwi-interacting RNA, Semen, Biology, QH426-470, TRFs, RRFs, [SDV.BDLR.RS]Life Sciences [q-bio]/Reproductive Biology/Sexual reproduction, 03 medical and health sciences, 0302 clinical medicine, [SDV.BBM.GTP]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Genomics [q-bio.GN], Gene expression, Embryonic morphogenesis, Testis, medicine, Genetics, Animals, Testicular parenchyma sperm, Molecular Biology, [SDV.BDD.GAM]Life Sciences [q-bio]/Development Biology/Gametogenesis, reproductive and urinary physiology, 030304 developmental biology, Secretome, Epididymis, 0303 health sciences, 030219 obstetrics & reproductive medicine, urogenital system, Research, PiRNA, Embryogenesis, SncRNAs, [SDV.BDLR]Life Sciences [q-bio]/Reproductive Biology, [SDV.BBM.BM]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Molecular biology, Bovine, Epididymis location, Sperm, Spermatozoa, Cell biology, Sperm epigenetics, medicine.anatomical_structure, RNA, Small Untranslated, Cattle, MiRNA, Spermatogenesis
Abstract

Background During epididymal transit, spermatozoa go through several functional maturation steps, resulting from interactions with epididymal secretomes specific to each region. In particular, the sperm membrane is under constant remodeling, with sequential attachment and shedding of various molecules provided by the epididymal lumen fluid and epididymosomes, which also deliver sncRNA cargo to sperm. As a result, the payload of sperm sncRNAs changes during the transit from the epididymis caput to the cauda. This work was designed to study the dynamics of cattle sperm sncRNAs from spermatogenesis to final maturation. Results Comprehensive catalogues of sperm sncRNAs were obtained from testicular parenchyma, epididymal caput, corpus and cauda, as well as ejaculated semen from three Holstein bulls. The primary cattle sncRNA sperm content is markedly remodeled as sperm mature along the epididymis. Expression of piRNAs, which are abundant in testis parenchyma, decreases dramatically at epididymis. Conversely, sperm progressively acquires miRNAs, rsRNAs, and tsRNAs along epididymis, with regional specificities. For instance, miRNAs and tsRNAs are enriched in epididymis cauda and ejaculated sperm, while rsRNA expression peaks at epididymis corpus. In addition, epididymis corpus contains mainly 20 nt long piRNAs, instead of 30 nt in all other locations. Beyond the bulk differences in abundance of sncRNAs classes, K-means clustering was performed to study their spatiotemporal expression profile, highlighting differences in specific sncRNAs and providing insights into their putative biological role at each maturation stage. For instance, Gene Ontology analyses using miRNA targets highlighted enriched processes such as cell cycle regulation, response to stress and ubiquitination processes in testicular parenchyma, protein metabolism in epididymal sperm, and embryonic morphogenesis in ejaculated sperm. Conclusions Our findings confirm that the sperm sncRNAome does not simply reflect a legacy of spermatogenesis. Instead, sperm sncRNA expression shows a remarkable level of plasticity resulting probably from the combination of multiple factors such as loss of the cytoplasmic droplet, interaction with epididymosomes, and more surprisingly, the putative in situ production and/or modification of sncRNAs by sperm. Given the suggested role of sncRNA in epigenetic trans-generational inheritance, our detailed spatiotemporal analysis may pave the way for a study of sperm sncRNAs role in embryo development.

Published
2021

6. Conceptus-induced, interferon tau-dependent gene expression in bovine endometrial epithelial and stromal cells†

Author

Daniel J Mathew, Susanta K. Behura, Gilles Charpigny, Lindsay F Grose, I. Martin Sheldon, Patrick Lonergan, James G. Cronin, Heather L Chaney, Thomas E. Spencer, West Virginia University [Morgantown], Biologie de la Reproduction, Environnement, Epigénétique & Développement (BREED), Université Paris-Saclay-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-École nationale vétérinaire d'Alfort (ENVA)-Université de Versailles Saint-Quentin-en-Yvelines (UVSQ), University of Missouri System, Swansea University, University College Dublin [Dublin] (UCD), and University of Tennessee System

Subjects
0301 basic medicine, Stromal cell, interferon tau, Pregnancy Proteins, Biology, Endometrium, 03 medical and health sciences, 0302 clinical medicine, Gene expression, medicine, Animals, Conceptus, Embryo Implantation, endometrium, Receptors, Ghrelin, Fibroblast, Sheep, 030219 obstetrics & reproductive medicine, RNA-Binding Proteins, Epithelial Cells, [SDV.BDLR]Life Sciences [q-bio]/Reproductive Biology, Cell Biology, General Medicine, Fibroblasts, Embryo, Mammalian, Coculture Techniques, Cell biology, Interferon tau, 030104 developmental biology, medicine.anatomical_structure, Gene Expression Regulation, conceptus, Reproductive Medicine, Interferon Type I, Cattle, Female, stromal, Stromal Cells, IRF8, Signal transduction, epithelium, transcriptome
Abstract

Bovine endometrium consists of epithelial and stromal cells that respond to conceptus interferon tau (IFNT), the maternal recognition of pregnancy (MRP) signal, by increasing expression of IFN-stimulated genes (ISGs). Endometrial epithelial and stromal-cell-specific ISGs are largely unknown but hypothesized to have essential functions during pregnancy establishment. Bovine endometrial epithelial cells were cultured in inserts above stromal fibroblast (SF) cells for 6 h in medium alone or with IFNT. The epithelial and SF transcriptomic response was analyzed separately using RNA sequencing and compared to a list of 369 DEGs recently identified in intact bovine endometrium in response to elongating bovine conceptuses and IFNT. Bovine endometrial epithelial and SF shared 223 and 70 DEGs in common with the list of 369 endometrial DEGs. Well-known ISGs identified in the epithelial and SF were ISG15, MX1, MX2, and OAS2. DEGs identified in the epithelial but not SF included a number of IRF molecules (IRF1, IRF2, IRF3, and IRF8), mitochondria SLC transporters (SLC25A19, SLC25A28, and SLC25A30), and a ghrelin receptor. Expression of ZC3HAV1, an anti-retroviral gene, increased specifically within the SF. Gene ontology analysis identified the type I IFN signaling pathway and activation of nuclear factor kappa B transcription factors as biological processes associated with the epithelial cell DEGs. This study has identified biologically relevant IFNT-stimulated genes within specific endometrial cell types. The findings provide critical information regarding the effects of conceptus IFNT on specific endometrial compartments during early developmental processes in cattle.Summary sentenceConceptus-induced, interferon-tau-dependent genes were identified within bovine endometrial epithelial and stromal fibroblast cells providing critical information regarding conceptus-induced modifications to specific endometrial compartments in cattle.

Published
2020

7. Heterochromatin establishment during early mammalian development is regulated by pericentromeric RNA and characterized by non-repressive H3K9me3

Author

Juan M. Vaquerizas, Maria-Elena Torres-Padilla, Thomas Jenuwein, Elias R. Ruiz-Morales, Nathalie Beaujean, Xiaoyu Liu, Vincent Brochard, Sriharsa Pradhan, Stéphanie Le Gras, Hang Gyeong Chin, André Eid, Vishnu S. Udayakumar, Diego Rodriguez-Terrones, Quirze Rovira, Chenfei Wang, Adam Burton, Shaorong Gao, Carmen Galán, Kai Kruse, Institute of Epigenetics and Stem Cells (IES), Helmholtz Zentrum Munchen, München, Germany, Biologie de la Reproduction, Environnement, Epigénétique & Développement (BREED), Université Paris-Saclay-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-École nationale vétérinaire d'Alfort (ENVA)-Université de Versailles Saint-Quentin-en-Yvelines (UVSQ), University of Extremadura, Inmunologia y Genetica Aplicada, Institut de Génétique et de Biologie Moléculaire et Cellulaire (IGBMC), Université de Strasbourg (UNISTRA)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Jiangsu Vocational College of Agriculture and Forestry, State Key Laboratory of Genetic Engineering, School of Life Sciences, Fudan-SJTU-Nottingham Plant Biotechnology R&D Center, Morgan-Tan International Center for Life Sciences, Fudan University [Shanghai], New England Biolabs, Max Planck Institute for Molecular Biomedicine, Max-Planck-Gesellschaft, Institut cellule souche et cerveau (U846 Inserm - UCBL1), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Institut National de la Recherche Agronomique (INRA), Max Planck Institute of Immunobiology and Epigenetics (MPI-IE), University Hospital of the Ludwig-Maximilians University, Biologie du développement et reproduction (BDR), École nationale vétérinaire d'Alfort (ENVA)-Institut National de la Recherche Agronomique (INRA)-Centre National de la Recherche Scientifique (CNRS), Institut cellule souche et cerveau / Stem Cell and Brain Research Institute (U1208 Inserm - UCBL1 / SBRI), Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Institut National de la Santé et de la Recherche Médicale (INSERM), Department of Industrial Microbiology and Biotechnology, Faculty of Biology and Environmental Protection, University of Lodz, and University of Lódź

Subjects
Male, Heterochromatin, [SDV]Life Sciences [q-bio], Centromere, epigenetic reprogramming, Embryonic Development, [SDV.BC]Life Sciences [q-bio]/Cellular Biology, Biology, Methylation, Article, Epigenesis, Genetic, H3K9me3, Histones, Mice, 03 medical and health sciences, 0302 clinical medicine, Gene expression, Animals, Constitutive heterochromatin, [SDV.BDD]Life Sciences [q-bio]/Development Biology, 030304 developmental biology, Epigenesis, Cell Nucleus, 0303 health sciences, Zygote, Pronucleus, fungi, Cell Biology, pre-implantation embryo, Embryo, Mammalian, Chromatin, Cell biology, Mice, Inbred C57BL, 030220 oncology & carcinogenesis, Mice, Inbred CBA, RNA, Female, Reprogramming
Abstract

Following fertilization in mammals, the gametes are reprogrammed to create a totipotent zygote, a process that involves de novo establishment of chromatin domains. A major feature occurring during preimplantation development is the dramatic remodelling of constitutive heterochromatin, although the functional relevance of this is unknown. Here, we show that heterochromatin establishment relies on the stepwise expression and regulated activity of SUV39H enzymes. Enforcing precocious acquisition of constitutive heterochromatin results in compromised development and epigenetic reprogramming, which demonstrates that heterochromatin remodelling is essential for natural reprogramming at fertilization. We find that de novo H3K9 trimethylation (H3K9me3) in the paternal pronucleus after fertilization is catalysed by SUV39H2 and that pericentromeric RNAs inhibit SUV39H2 activity and reduce H3K9me3. De novo H3K9me3 is initially non-repressive for gene expression, but instead bookmarks promoters for compaction. Overall, we uncover the functional importance for the restricted transmission of constitutive heterochromatin during reprogramming and a non-repressive role for H3K9me3.Burton et al. show that H3K9me3 deposition catalysed by SUV39H2 and regulated by pericentromeric RNAs in the mouse paternal pronucleus does not suppress gene expression, but bookmarks promoters for compaction.

Published
2020

8. Investigating the role of BCAR4 in ovarian physiology and female fertility by genome editing in rabbit

Author

Rozenn Dalbies-Tran, Maud Peyny, Peggy Jarrier-Gaillard, Sébastien Lavillatte, Laurent Boulanger, Nathalie Daniel, Pascal Papillier, Véronique Duranthon, Geneviève Jolivet, Véronique Cadoret, Danielle Monniaux, Nathalie Peynot, Physiologie de la reproduction et des comportements [Nouzilly] (PRC), Institut Français du Cheval et de l'Equitation [Saumur]-Université de Tours-Centre National de la Recherche Scientifique (CNRS)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Biologie de la Reproduction, Environnement, Epigénétique & Développement (BREED), Université Paris-Saclay-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-École nationale vétérinaire d'Alfort (ENVA)-Université de Versailles Saint-Quentin-en-Yvelines (UVSQ), Plateforme d'Infectiologie Expérimentale (PFIE), Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), CHU Bretonneau, Médecine et Biologie de la Reproduction-CECOS, INRAE, Agence Nationale de la Biomédecine, Institut Français du Cheval et de l'Equitation [Saumur]-Université de Tours (UT)-Centre National de la Recherche Scientifique (CNRS)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), and École nationale vétérinaire d'Alfort (ENVA)-Université de Versailles Saint-Quentin-en-Yvelines (UVSQ)-Université Paris-Saclay-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)

Subjects
0301 basic medicine, rabbits, media_common.quotation_subject, [SDV]Life Sciences [q-bio], Embryonic Development, Gene Expression, Reproductive biology, lcsh:Medicine, Ovary, Biology, Oogenesis, Article, Andrology, 03 medical and health sciences, 0302 clinical medicine, Ovarian Follicle, Follicular phase, medicine, Animals, Blastocyst, Ovarian follicle, lcsh:Science, Ovulation, genome, media_common, Gene Editing, fertility, Multidisciplinary, lcsh:R, Functional genomics, Oocyte, 030104 developmental biology, medicine.anatomical_structure, 030220 oncology & carcinogenesis, ovulation, Female, RNA, Long Noncoding, lcsh:Q, Folliculogenesis, Anti-estrogen
Abstract

Breast Cancer Anti-estrogen Resistance 4 (BCAR4) was previously characterised in bovine species as a gene preferentially expressed in oocytes, whose inhibition is detrimental to in vitro embryo development. But its role in oogenesis, folliculogenesis and globally fertility in vivo remains unknown. Because the gene is not conserved in mice, rabbits were chosen for investigation of BCAR4 expression and function in vivo. BCAR4 displayed preferential expression in the ovary compared to somatic organs, and within the ovarian follicle in the oocyte compared to somatic cells. The transcript was detected in follicles as early as the preantral stage. Abundance decreased throughout embryo development until the blastocyst stage. A lineage of genome-edited rabbits was produced; BCAR4 expression was abolished in follicles from homozygous animals. Females of wild-type, heterozygous and homozygous genotypes were examined for ovarian physiology and reproductive parameters. Follicle growth and the number of ovulations in response to hormonal stimulation were not significantly different between genotypes. Following insemination, homozygous females displayed a significantly lower delivery rate than their heterozygous counterparts (22 ± 7% vs 71 ± 11% (mean ± SEM)), while prolificacy was 1.8 ± 0.7 vs 6.0 ± 1.4 kittens per insemination. In conclusion, BCAR4 is not essential for follicular growth and ovulation but it contributes to optimal fertility in rabbits.

Published
2020

9. Ovine fetal testis stage-specific sensitivity to environmental chemical mixtures

Author

Richard G Lea, Beatrice Mandon-Pépin, Benoit Loup, Elodie Poumerol, Luc Jouneau, Biola F Egbowon, Adelle Bowden, Corinne Cotinot, Laura Purdie, Zulin Zhang, Paul A Fowler, Kevin D Sinclair, University of Nottingham, UK (UON), Biologie de la Reproduction, Environnement, Epigénétique & Développement (BREED), École nationale vétérinaire - Alfort (ENVA)-Université de Versailles Saint-Quentin-en-Yvelines (UVSQ)-Université Paris-Saclay-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), The James Hutton Institute, University of Aberdeen, and European Project: 212885,EC:FP7:ENV,FP7-ENV-2007-1,REEF(2008)

Subjects
Male, Embryology, Sheep, Sewage, Obstetrics and Gynecology, [SDV.BDLR]Life Sciences [q-bio]/Reproductive Biology, Cell Biology, Fetus, Endocrinology, Reproductive Medicine, Pregnancy, Testis, environmental chemical mixtures, Animals, Humans, stage-specific sensitivity, Female, Testosterone, [SDV.BDD]Life Sciences [q-bio]/Development Biology, Sheep, Domestic
Abstract

International audience; Exposure of the fetal testis to numerous individual environmental chemicals (ECs) is frequently associated with dysregulated development, leading to impaired adult reproductive competence. However, ‘real-life’ exposure involves complex mixtures of ECs. Here we test the consequences, for the male fetus, of exposing pregnant ewes to EC mixtures derived from pastures treated with biosolids fertiliser (processed human sewage). Fetal testes from continuously exposed ewes were either unaffected at day 80 or exhibited a reduced area of testis immunostained for CYP17A1 protein at day 140. Fetal testes from day 140 pregnant ewes that were exposed transiently for 80-day periods during early (0–80 days), mid (30–110 days), or late (60–140 days) pregnancy had fewer Sertoli cells and reduced testicular area stained for CYP17A1. Male fetuses from ewes exposed during late pregnancy also exhibited reduced fetal body, adrenal and testis mass, anogenital distance, and lowered testosterone; collectively indicative of an anti-androgenic effect. Exposure limited to early gestation induced more testis transcriptome changes than observed for continuously exposed day 140 fetuses. These data suggest that a short period of EC exposure does not allow sufficient time for the testis to adapt. Consequently, testicular transcriptomic changes induced during the first 80 days of gestation may equate with phenotypic effects observed at day 140. In contrast, relatively fewer changes in the testis transcriptome in fetuses exposed continuously to ECs throughout gestation are associated with less severe consequences. Unless corrected by or during puberty, these differential effects would predictably have adverse outcomes for adult testicular function and fertility.

Published
2022

10. Absence of Testicular Estrogen Leads to Defects in Spermatogenesis and Increased Semen Abnormalities in Male Rabbits

Author

Aurélie Dewaele, Emilie Dujardin, Marjolaine André, Audrey Albina, Hélène Jammes, Frank Giton, Eli Sellem, Geneviève Jolivet, Eric Pailhoux, Maëlle Pannetier, Biologie de la Reproduction, Environnement, Epigénétique & Développement (BREED), École nationale vétérinaire - Alfort (ENVA)-Université de Versailles Saint-Quentin-en-Yvelines (UVSQ)-Université Paris-Saclay-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Institut Mondor de Recherche Biomédicale (IMRB), Institut National de la Santé et de la Recherche Médicale (INSERM)-IFR10-Université Paris-Est Créteil Val-de-Marne - Paris 12 (UPEC UP12), Hôpital Henri Mondor, ANR-09-GENM-0009,GENIDOV,Réseaux géniques impliqués dans la différenciation ovarienne chez la chèvre et la souris(2009), ANR-13-BSV2-0017,ARGONADS,Acide rétinoique et devenir des cellules germinales(2013), ANR-20-CE14-0022,ARDIGERM,Acide rétinoïque dans la différenciation des cellules germinales et la méiose(2020), BREED, INRAE, Génomique et biotechnologies végétales - Réseaux géniques impliqués dans la différenciation ovarienne chez la chèvre et la souris - - GENIDOV2009 - ANR-09-GENM-0009 - GENOM-BTV - VALID, Blanc 2013 - Acide rétinoique et devenir des cellules germinales - - ARGONADS2013 - ANR-13-BSV2-0017 - Blanc 2013 - VALID, and Acide rétinoïque dans la différenciation des cellules germinales et la méiose - - ARDIGERM2020 - ANR-20-CE14-0022 - AAPG2020 - VALID

Subjects
CYP19A1 knock-out, male fertility, testicular estrogens, epididymis, spermatogenesis, sperm maturation, DNA methylation, Male, Mammals, Estrogens, [SDV.BDLR]Life Sciences [q-bio]/Reproductive Biology, Semen, Testis, Genetics, Sperm Motility, Humans, Animals, Female, Rabbits, Genetics (clinical), [SDV.BDLR] Life Sciences [q-bio]/Reproductive Biology
Abstract

Estrogens are steroid hormones produced by the aromatization of androgens by the aromatase enzyme, encoded by the CYP19A1 gene. Although generally referred to as “female sex hormones”, estrogen is also produced in the adult testes of many mammals, including humans. To better understand the function of estrogens in the male, we used the rabbit model which is an important biomedical model. First, the expression of CYP19A1 transcripts was localized mainly in meiotic germ cells. Thus, testicular estrogen appears to be produced inside the seminiferous tubules. Next, the cells expressing ESR1 and ESR2 were identified, showing that estrogens could exert their function on post-meiotic germ cells in the tubules and play a role during sperm maturation, since ESR1 and ESR2 were detected in the cauda epididymis. Then, CRISPR/Cas9 CYP19A1−/− genetically modified rabbits were analyzed. CYP19A1−/− males showed decreased fertility with lower sperm count associated with hypo-spermatogenesis and lower spermatid number. Germ/sperm cell DNA methylation was unchanged, while sperm parameters were affected as CYP19A1−/− males exhibited reduced sperm motility associated with increased flagellar defects. In conclusion, testicular estrogens could be involved in the spermatocyte–spermatid transition in the testis, and in the acquisition of sperm motility in the epididymis.

Published
2022

11. Maternal age affects equine Day 8 embryo gene expression both in trophoblast and inner cell mass

Author

Michèle Dahirel, Yan Jaszczyszyn, Duranthon, Luc Jouneau, Wimel L, Nathalie Peynot, Pascale Chavatte-Palmer, Dubois C, Auclair-Ronzaud J, Catherine Archilla, Nathalie Daniel, E. Derisoud, Rachel Legendre, Biologie de la Reproduction, Environnement, Epigénétique & Développement (BREED), École nationale vétérinaire - Alfort (ENVA)-Université de Versailles Saint-Quentin-en-Yvelines (UVSQ)-Université Paris-Saclay-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Institut Français du Cheval et de l'Equitation [Saumur] (IFCE), Institut de Biologie Intégrative de la Cellule (I2BC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), Hub Bioinformatique et Biostatistique - Bioinformatics and Biostatistics HUB, Institut Pasteur [Paris] (IP)-Université Paris Cité (UPCité), This work was supported by the 'Institut Français du Cheval et de l’Equitation' (grant numbers CS_2018_23, 2018). The National Research Institute for Agriculture, Food and Environment (INRAE) department Animal Physiology and Breeding Systems also supported this research., ANR-11-INBS-0013,IFB (ex Renabi-IFB),Institut français de bioinformatique(2011), Université Paris-Saclay-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-École nationale vétérinaire d'Alfort (ENVA)-Université de Versailles Saint-Quentin-en-Yvelines (UVSQ), Institut Français du Cheval et de L'équitation (IFCE), Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), Station experimentale des Haras Nationaux [Plateau technique de Chamberet], Institut Français du Cheval et de l'Equitation, This work was supported by the 'Institut Français du Cheval et de l’Equitation'(grant numbers CS_2018_23, 2018). The National Research Institute for Agriculture, Food and Environment (INRAE) department Animal Physiology and Breeding Systems also supported this research., and DERISOUD, Emilie

Subjects
Male, [SDV.BA] Life Sciences [q-bio]/Animal biology, [SDV]Life Sciences [q-bio], Mare, Gene Expression, Biology, Breeding, [SDV.BDLR.RS]Life Sciences [q-bio]/Reproductive Biology/Sexual reproduction, Andrology, 03 medical and health sciences, 0302 clinical medicine, Gene expression, [SDV.BDD] Life Sciences [q-bio]/Development Biology, [SDV.BA.ZV]Life Sciences [q-bio]/Animal biology/Vertebrate Zoology, medicine, Genetics, Inner cell mass, Animals, Blastocyst, Horses, Gene, [SDV.BDD]Life Sciences [q-bio]/Development Biology, [SDV.BDLR] Life Sciences [q-bio]/Reproductive Biology, reproductive and urinary physiology, 030304 developmental biology, fertility, 0303 health sciences, 030219 obstetrics & reproductive medicine, [SDV.BDLR.RS] Life Sciences [q-bio]/Reproductive Biology/Sexual reproduction, [SDV.BA]Life Sciences [q-bio]/Animal biology, Embryogenesis, Trophoblast, [SDV.BDD.EO] Life Sciences [q-bio]/Development Biology/Embryology and Organogenesis, Embryo, [SDV.BDLR]Life Sciences [q-bio]/Reproductive Biology, trophoblast, Trophoblasts, Plant Breeding, medicine.anatomical_structure, [SDV.BDD.EO]Life Sciences [q-bio]/Development Biology/Embryology and Organogenesis, embryonic structures, RNA, [SDV.BA.ZV] Life Sciences [q-bio]/Animal biology/Vertebrate Zoology, Female, Embryo quality, Biotechnology, Maternal Age
Abstract

Background Increased embryo loss as mares become older is a major consideration for breeders as older animals are currently used for reproduction in the equine industry. Lower embryo quality has been pointed out as partly responsible for this reduced fertility. Here, the effect of mare’s age on blastocysts’ gene expression was explored. Day 8 post ovulation embryos were collected by uterine flushing from multiparous young (YM, 6-year-old, N = 5) and older (OM, > 10-year-old, N = 6) non-nursing Saddlebred mares, inseminated with the semen of one stallion. Pure (TE_part) or inner cell mass enriched (ICMandTE) trophoblast were obtained by embryo bisection and paired end, non-oriented RNA sequencing (Illumina, NextSeq500) was performed on each hemi-embryo. To discriminate gene expression in the ICM from that in the TE, deconvolution (DeMixT R package) was used on the ICMandTE dataset. Differential expression was analyzed (DESeq2) with embryo sex and diameter as cofactors using a false discovery rate < 0.05 cutoff. Functional annotation and classification of differentially expressed genes and gene set enrichment analysis were also performed. Results :Maternal aging did not affect embryo recovery rate, embryo diameter nor total RNA quantity but modified gene expression in equine D8 embryos. The expression of genes in the ICM seemed to be more altered by maternal aging than in the TE. In both compartments, the expression of genes involved in mitochondrial function, translation and transcription due to chromatin modification were disturbed by maternal age. Mitosis, signaling and adhesion pathways and embryo development were particularly decreased in the ICM hemi-embryos from old mares. Finally, in TE, ion movement related genes were affected. Conclusions :This is the first study showing an effect of maternal age on gene expression in the equine blastocyst at Day 8 post ovulation. Maternal age, even for mares as young as 10 years old, disturbs mitosis, translation and transcription, cell signaling and adhesion as well as mitochondrial function and cell commitment of horse embryos. These perturbations may affect further embryo development and contribute to decreased fertility due to aging.

Published
2021

12. DYRK1A Overexpression in Mice Downregulates the Gonadotropic Axis and Disturbs Early Stages of Spermatogenesis

Author

Nathalie Janel, Antonin Lamaziere, Chrystèle Racine, François Vialard, Rodolphe Dard, Valérie Serazin, Nadim Kassis, Estelle Parizot, Farah Ghieh, Nathalie di Clemente, Manon Moreau, Leslie Brehier, Unité de Biologie Fonctionnelle et Adaptative (BFA (UMR_8251 / U1133)), Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université de Paris (UP), Biologie de la Reproduction, Environnement, Epigénétique & Développement (BREED), Université Paris-Saclay-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-École nationale vétérinaire d'Alfort (ENVA)-Université de Versailles Saint-Quentin-en-Yvelines (UVSQ), CHI Poissy-Saint-Germain, Centre de Recherche Saint-Antoine (CR Saint-Antoine), Sorbonne Université (SU)-Institut National de la Santé et de la Recherche Médicale (INSERM)-CHU Saint-Antoine [AP-HP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU), Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université Paris Cité (UPCité), École nationale vétérinaire - Alfort (ENVA)-Université de Versailles Saint-Quentin-en-Yvelines (UVSQ)-Université Paris-Saclay-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Centre de Recherche Saint-Antoine (CRSA), Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Sorbonne Université (SU), and Gestionnaire, HAL Sorbonne Université 5

Subjects
Male, Genetically modified mouse, Infertility, Down syndrome, medicine.medical_specialty, DYRK1A, Transgene, [SDV.GEN] Life Sciences [q-bio]/Genetics, Protein Serine-Threonine Kinases, QH426-470, Article, Mice, 03 medical and health sciences, 0302 clinical medicine, Hypogonadotropic hypogonadism, Internal medicine, Testis, Genetics, medicine, Animals, Spermatogenesis, Zebrafish, Infertility, Male, Genetics (clinical), 030304 developmental biology, 0303 health sciences, [SDV.GEN]Life Sciences [q-bio]/Genetics, biology, Hypogonadism, Protein-Tyrosine Kinases, biology.organism_classification, medicine.disease, Up-Regulation, Mice, Inbred C57BL, Endocrinology, infertility, 030217 neurology & neurosurgery
Abstract

International audience; Down syndrome (DS) is the most common chromosomal disorder. It is responsible for intellectual disability (ID) and several medical conditions. Although men with DS are thought to be infertile, some spontaneous paternities have been reported. The few studies of the mechanism of infertility in men with DS are now dated. Recent research in zebrafish has indicated that overexpression of DYRK1A (the protein primarily responsible for ID in DS) impairs gonadogenesis at the embryonic stage. To better ascertain DYRK1A’s role in infertility in DS, we investigated the effect of DYRK1A overexpression in a transgenic mouse model. We found that overexpression of DYRK1A impairs fertility in transgenic male mice. Interestingly, the mechanism in mice differs slightly from that observed in zebrafish but, with disruption of the early stages of spermatogenesis, is similar to that seen in humans. Unexpectedly, we observed hypogonadotropic hypogonadism in the transgenic mice.

Published
2021

13. Sex Chromosomes and Master Sex-Determining Genes in Turtles and Other Reptiles

Author

Dominique Thépot, Biologie de la Reproduction, Environnement, Epigénétique & Développement (BREED), and Université Paris-Saclay-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-École nationale vétérinaire d'Alfort (ENVA)-Université de Versailles Saint-Quentin-en-Yvelines (UVSQ)

Subjects
0106 biological sciences, Male, homologous genes, Review, Biology, QH426-470, 010603 evolutionary biology, 01 natural sciences, Genome, DNA sequencing, RAD-seq, Genetic Sex Determination (GSD), Transcriptome, 03 medical and health sciences, Molecular level, Genetics, Animals, Gene, Genetics (clinical), 030304 developmental biology, 0303 health sciences, Sex Chromosomes, RNA, [SDV.BDLR]Life Sciences [q-bio]/Reproductive Biology, Sex Determination Processes, snakes, Turtles, lizards, Testis determining factor, Evolutionary biology, genome coverage, squamates, Identification (biology), Female
Abstract

International audience; Among tetrapods, the well differentiated heteromorphic sex chromosomes of birds and mammals have been highly investigated and their master sex-determining (MSD) gene, Dmrt1 and SRY, respectively, have been identified. The homomorphic sex chromosomes of reptiles have been the least studied, but the gap with birds and mammals has begun to fill. This review describes our current knowledge of reptilian sex chromosomes at the cytogenetic and molecular level. Most of it arose recently from various studies comparing male to female gene content. This includes restriction site-associated DNA sequencing (RAD-Seq) experiments in several male and female samples, RNA sequencing and identification of Z- or X-linked genes by male/female comparative transcriptome coverage, and male/female transcriptomic or transcriptome/genome substraction approaches allowing the identification of Y- or W-linked transcripts. A few putative master sex-determining (MSD) genes have been proposed, but none has been demonstrated yet. Lastly, future directions in the field of reptilian sex chromosomes and their MSD gene studies are considered.

Published
2021

14. Moderate differences in plasma leptin in mares have no effect on either the amino acid or the fatty acid composition of the uterine fluid

Author

Vladimir Milojevic, Barbara Drews, Katrin Giller, Pieter Giesbertz, Hannelore Daniel, Susanne E. Ulbrich, Cédric Dubois, Rupert M. Bruckmaier, Morgane Robles, A. Rudolf Vegas, L. Wimel, Pascale Chavatte-Palmer, Institute Agricultural Science (ETH Zurich), Biologie de la Reproduction, Environnement, Epigénétique & Développement (BREED), École nationale vétérinaire d'Alfort (ENVA)-Université de Versailles Saint-Quentin-en-Yvelines (UVSQ)-Université Paris-Saclay-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Institut Français du Cheval et de L'équitation (IFCE), Institute of Food Nutrition and Health (ETH), Technische University Munich, VetSuisse Faculty, Clinic of Reproductive Medicine, Genetics and Functional Genomics Group, Universität Zürich [Zürich] = University of Zurich (UZH)-Universität Zürich [Zürich] = University of Zurich (UZH)-Genetics and Functional Genomics Group, and Universität Zürich [Zürich] = University of Zurich (UZH)-Universität Zürich [Zürich] = University of Zurich (UZH)

Subjects
Hypothalamo-Hypophyseal System, medicine.medical_specialty, obesity, Linoleic acid, Pituitary-Adrenal System, leptin, fatty acids, Plasma, 03 medical and health sciences, chemistry.chemical_compound, 0302 clinical medicine, Internal medicine, medicine, Animals, Horses, Amino Acids, ComputingMilieux_MISCELLANEOUS, Retrospective Studies, 030304 developmental biology, chemistry.chemical_classification, 0303 health sciences, Body condition, 030219 obstetrics & reproductive medicine, [SDV.BA.MVSA]Life Sciences [q-bio]/Animal biology/Veterinary medicine and animal Health, Adiponectin, uterus, Equine, Reproduction, Leptin, Fatty acid, 3. Good health, Amino acid, horse, Glutamine, Endocrinology, chemistry, Uterine fluid, Metabolic hormones, Female, Stearic acid, FOS: Medical biotechnology, Horse, Hormone
Abstract

Female mammalian reproductive functions are closely linked to body condition and metabolic status. Energy homeostasis is regulated by endocrine hormones such as insulin, IGF-I, leptin, and adiponectin via the hypothalamic-pituitary-adrenal axis. These metabolic hormones and their receptors are also expressed in reproductive tissues and the embryo. We investigated the relationship between circulating leptin and the fatty acid (FA) and amino acid (AA) composition of the equine uterine fluid (UF) and peripheral blood plasma (BP) by using a mass spectrometry-based approach. UF and BP were collected from ten broodmares on days 6 and 7 post ovulation, respectively. The mares were retrospectively assigned to two groups according to their BP leptin concentrations (high leptin [> 1.6 ng/mL] versus low leptin [, Journal of Equine Veterinary Science, 109

Published
2021

15. Meloxicam administration in the management of postoperative pain and inflammation associated with caesarean section in beef heifers: Evaluation of reproductive parameters

Author

Thomas Cardot, B. Grimard, Nicolas Roch, Vincent Mauffré, Fabienne Constant, Andrew Ponter, Guillaume Belbis, Nicolas Nehlig, Arnaud Bohy, Vincent Plassard, Laurence Guilbert-Julien, Sandrine Bernard, Biologie de la Reproduction, Environnement, Epigénétique & Développement (BREED), Université Paris-Saclay-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-École nationale vétérinaire d'Alfort (ENVA)-Université de Versailles Saint-Quentin-en-Yvelines (UVSQ), École nationale vétérinaire d'Alfort (ENVA), Groupement Technique Vétérinaire (GTV), Boehringer Ingelheim Animal Health, Partenaires INRAE, and Laboratoire National de Contrôle des Reproducteurs (LNCR)

Subjects
medicine.medical_specialty, 040301 veterinary sciences, medicine.medical_treatment, Cattle Diseases, Ice calving, Culling, Meloxicam, 0403 veterinary science, C-section, Food Animals, Pregnancy, Retained placenta, Animals, Medicine, Caesarean section, Non-steroidal anti-inflammatory drug, Small Animals, Survival analysis, Inflammation, 2. Zero hunger, Pain, Postoperative, [SDV.BA.MVSA]Life Sciences [q-bio]/Animal biology/Veterinary medicine and animal Health, Cesarean Section, Equine, business.industry, Obstetrics, Incidence (epidemiology), Reproduction, 0402 animal and dairy science, 04 agricultural and veterinary sciences, medicine.disease, 040201 dairy & animal science, Pain management, 3. Good health, Beef heifers, Pregnancy rate, Cattle, Female, Animal Science and Zoology, business, medicine.drug
Abstract

International audience; Post-operative pain and inflammation are normal physiological reactions to caesarean section. Their management in cattle have rarely been investigated. This surgical procedure negatively affects reproductive function with, for example, a reduction in fertility resulting in an increase in calving interval. In this multicenter clinical trial, the objective was to evaluate the impact on reproductive performance of meloxicam injected before caesarean section to manage post-operative pain and inflammation. Meloxicam is a non-steroidal anti-inflammatory drug. One hundred and twenty-seven Charolais heifers (n = 127) were recruited from 47 farms in six French veterinary practices in the Burgundy region. The heifers underwent a non-elective standardized caesarean section operation. Heifers were randomly assigned to one of two groups: meloxicam (n = 66), intravenous meloxicam injection before surgery, or control (n = 61). Reproductive performance and health information were recorded from the time of the caesarean section to the next calving or to culling. In our study, meloxicam administration before caesarean section had no effect on the incidence of retained placenta (18.2% of treated vs 25.0% of control cows, p = 0.35). The pregnancy rate was higher in treated than in control cows (83.1% vs 67.8%, p = 0.04 after multivariate analysis) and a survival analysis showed that the median calving interval was 35 days shorter in the meloxicam (t50% = 417 days) compared to the control group (t50% = 452 days, p = 0.05). A trend was also observed for culling rate to be lower in treated (4.7%) compared to control cows (13.3%, p = 0.09). In conclusion, this study suggests that there is a beneficial effect of meloxicam administration before caesarean section on reproductive performance in Charolais heifers.

Published
2021

16. Fetal estrogens are not involved in sex determination but critical for early ovarian differentiation in rabbits

Author

Eloïse Airaud, Aurélie Dewaele, Laurent Boulanger, Nathalie Daniel-Carlier, Frank Giton, Beatrice Mandon-Pepin, Maëlle Pannetier, Geneviève Jolivet, Cloé Pierson, Eric Pailhoux, Erwana Harscoët, Nathalie Daniel, Biologie de la Reproduction, Environnement, Epigénétique & Développement (BREED), École nationale vétérinaire d'Alfort (ENVA)-Université de Versailles Saint-Quentin-en-Yvelines (UVSQ)-Université Paris-Saclay-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), École nationale vétérinaire - Alfort (ENVA)-Université de Versailles Saint-Quentin-en-Yvelines (UVSQ)-Université Paris-Saclay-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Institut Mondor de Recherche Biomédicale (IMRB), Institut National de la Santé et de la Recherche Médicale (INSERM)-IFR10-Université Paris-Est Créteil Val-de-Marne - Paris 12 (UPEC UP12), ANR-09-GENM-0009,GENIDOV,Réseaux géniques impliqués dans la différenciation ovarienne chez la chèvre et la souris(2009), ANR-13-BSV2-0017,ARGONADS,Acide rétinoique et devenir des cellules germinales(2013), IMRB - 'Biologie du système neuromusculaire' [Créteil] (U955 Inserm - UPEC), École nationale vétérinaire - Alfort (ENVA)-Institut Mondor de Recherche Biomédicale (IMRB), Institut National de la Santé et de la Recherche Médicale (INSERM)-IFR10-Université Paris-Est Créteil Val-de-Marne - Paris 12 (UPEC UP12)-Institut National de la Santé et de la Recherche Médicale (INSERM)-IFR10-Université Paris-Est Créteil Val-de-Marne - Paris 12 (UPEC UP12), Hôpital Henri Mondor, This study was supported by ANR grants (GENIDOV:ANR-09-GENM-009, ARGONADS: ANR-13-BSV2-0017, ARDIGERM: ANR-2020-CE14), and the CELPHEDIA infrastructure (CELPHEDIA France, 2017)., ANR-20-CE14-0022,ARDIGERM,Acide rétinoïque dans la différenciation des cellules germinales et la méiose(2020), Jolivet, Geneviève, Génomique et biotechnologies végétales - Réseaux géniques impliqués dans la différenciation ovarienne chez la chèvre et la souris - - GENIDOV2009 - ANR-09-GENM-0009 - GENOM-BTV - VALID, Blanc 2013 - Acide rétinoique et devenir des cellules germinales - - ARGONADS2013 - ANR-13-BSV2-0017 - Blanc 2013 - VALID, and Acide rétinoïque dans la différenciation des cellules germinales et la méiose - - ARDIGERM2020 - ANR-20-CE14-0022 - AAPG2020 - VALID

Subjects
Anti-Mullerian Hormone, Sex Differentiation, Somatic cell, [SDV]Life Sciences [q-bio], sex determination, AROMATASE, 0302 clinical medicine, INDEL Mutation, Ovarian Follicle, TALEN, Testosterone, Aromatase, Antrum, [SDV.BDD]Life Sciences [q-bio]/Development Biology, media_common, 0303 health sciences, 030219 obstetrics & reproductive medicine, Estradiol, biology, Gene Expression Regulation, Developmental, Cell Differentiation, Phenotype, medicine.anatomical_structure, Female, Cytochrome P450 Family 19, Rabbits, AcademicSubjects/MED00250, Research Article, Ovulation, endocrine system, Gonad, media_common.quotation_subject, rabbit, Andrology, 03 medical and health sciences, [SDV.BDD] Life Sciences [q-bio]/Development Biology, medicine, Animals, CYP19A1, Gonads, Ovarian reserve, Cell Proliferation, 030304 developmental biology, Fetus, Sexual differentiation, Gene Expression Profiling, Estrogens, Sex Determination Processes, CYP26A1, biology.protein, ovary
Abstract

AROMATASE, encoded by the CYP19A1 gene, is the cytochrome enzyme responsible for the synthesis of estrogens in vertebrates. In most mammals a peak of expression of the CYP19A1 gene occurs in the fetal XX gonad when sexual differentiation starts up. To elucidate the role of this peak, we produced 3 lines of TALEN genetically edited CYP19A1 KO rabbits, that were void of any production of estradiol. All KO XX rabbits developed as females, with aberrantly small sized ovaries at adulthood, an almost empty reserve of primordial follicles and very few large antrum follicles. Ovulation never occurred. Our histological, immunohistological and transcriptomic analyses showed that the surge of estradiol in the XX fetal rabbit gonad is dispensable for its determination as an ovary, or for meiosis. However, it is mandatory for the high proliferation and differentiation of both somatic and germ cells, and consequently for the establishment of the ovarian reserve.

Published
2021

17. Algorithms predicting gestational stage from the maternal steroid metabolome of mares

Author

Alan J Conley, Paul R. Shorten, Pascale Chavatte-Palmer, Erin L. Legacki, AGRESEARCH RUAKURA RESEARCH CENTRE HAMILTON NZL, Partenaires IRSTEA, Institut national de recherche en sciences et technologies pour l'environnement et l'agriculture (IRSTEA)-Institut national de recherche en sciences et technologies pour l'environnement et l'agriculture (IRSTEA), Department of Population Health and Reproduction Davis, California, University of California [Davis] (UC Davis), University of California-University of California, Biologie de la Reproduction, Environnement, Epigénétique & Développement (BREED), Université Paris-Saclay-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-École nationale vétérinaire d'Alfort (ENVA)-Université de Versailles Saint-Quentin-en-Yvelines (UVSQ), and IFCE Foetalim Project

Subjects
endocrine system, Pregnancy Tests, Endocrinology, Diabetes and Metabolism, Population, Dehydroepiandrosterone, Datasets as Topic, 030209 endocrinology & metabolism, Gestational Age, Biology, progesterone, 03 medical and health sciences, 0302 clinical medicine, Endocrinology, Pregnancy, Tandem Mass Spectrometry, Prenatal Diagnosis, medicine, Animals, Horses, education, reproductive and urinary physiology, progestagens, 030304 developmental biology, 2. Zero hunger, 0303 health sciences, Fetus, education.field_of_study, [SDV.BA.MVSA]Life Sciences [q-bio]/Animal biology/Veterinary medicine and animal Health, [SDV.BA]Life Sciences [q-bio]/Animal biology, Gestational age, Models, Theoretical, medicine.disease, Breed, Gonadotropin secretion, horse, Metabolome, Gestation, Feasibility Studies, Pregnancy, Animal, Female, Steroids, Algorithm, Algorithms, Chromatography, Liquid
Abstract

Hormone secretion by the maternal ovaries, trophoblast/placenta and fetus occurs sequentially, creating distinct steroid metabolomic ‘signatures’ in systemic blood of pregnant mares that vary with gestational stage. Algorithms were developed to predict the gestational day (GD) from the maternal steroid metabolome (nine steroids; pregnenolone (P5), progesterone (P4), 5α-dihydroprogesterone (DHP), 17α-hydroxyprogesterone, allopregnanolone, 20α-hydroxy-DHP, 3β,20α-dihydroxy-DHP, DHEA and androstenedione) determined by liquid chromatography-tandem mass spectrometry (LC-MS/MS) of eight thoroughbred mares sampled longitudinally throughout pregnancy. A physiologically based model was developed to infer rates of steroid secretion during chorionic gonadotropin secretion, the luteo-placental shift and by the equine feto-placenta unit, demonstrating more variability in P5 and DHP than P4. The average of four empirical models, using nine steroids to predict GD, was calibrated (five mares, R2 = 0.94, RMSE = 20 days) and validated (three mares, R2 = 0.84, RMSE = 32 days). Validation performance was improved using paired samples taken 14 or 30 days apart (RMSE = 29 and 19 days, respectively). A second validation used an independent dataset (single serum samples from 56 mixed breed mares, RMSE = 79 days) and an additional longitudinal subset from the same population sampled monthly throughout gestation (seven mares, RMSE = 42 days). Again, using paired samples improved model performance (RMSE = 32.5 days). Despite less predictive performance of the mixed breed than the thoroughbred datasets, these models demonstrate the feasibility and potential for using maternal steroid metabolomic algorithms to estimate the stage of gestation in pregnant mares and perhaps monitor fetal development.

Published
2021

18. Pregnancy and placental development in horses: an update

Author

Morgane Robles, E. Derisoud, Pascale Chavatte-Palmer, Biologie de la Reproduction, Environnement, Epigénétique & Développement (BREED), and École nationale vétérinaire d'Alfort (ENVA)-Université de Versailles Saint-Quentin-en-Yvelines (UVSQ)-Université Paris-Saclay-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)

Subjects
Gerontology, medicine.medical_specialty, placenta, Offspring, [SDV]Life Sciences [q-bio], Biology, Fetal Development, 03 medical and health sciences, 0302 clinical medicine, Endocrinology, Food Animals, Internal medicine, medicine, Animals, Humans, implantation, Horses, Progesterone, ComputingMilieux_MISCELLANEOUS, 030304 developmental biology, 0303 health sciences, Pregnancy, 030219 obstetrics & reproductive medicine, [SDV.BA.MVSA]Life Sciences [q-bio]/Animal biology/Veterinary medicine and animal Health, DOHaD, Maternal Nutritional Physiological Phenomena, medicine.disease, Placentation, horse, Gestation, Female, Animal Science and Zoology, pregnancy
Abstract

Horses have been domesticated by man and historical information mostly associates horses with men. Nowadays, however, horse riding is essentially practised by women. Women are also very much involved in equine sciences, with a large contribution to the understanding of foetoplacental development. While highlighting the work of female scientists, this review describes the recent advances in equine foetoplacental studies, focussing on data obtained by new generation sequencing and progress on the understanding of the role of placental progesterone metabolites throughout gestation. A second emphasis is made on foetal programming, a currently very active field, where the importance of maternal nutrition, mare management or the use of embryo technologies has been shown to induce long term effects in the offspring that might affect progeny's performance. Finally, new perspectives for the study of equine pregnancy are drawn, that will rely on new methodologies applied to molecular explorations and imaging.

Published
2021

19. Hepatic global DNA Hypomethylation Phenotype in Rainbow Trout Fed Diets Varying in Carbohydrate to Protein Ratio

Author

Lucie Marandel, Christine Burel, Jésabel Laithier, Vincent Veron, Cécile Heraud, Audrey Prézelin, Jingwei Liu, Stéphane Panserat, Nutrition, Métabolisme, Aquaculture (NuMéA), Université de Pau et des Pays de l'Adour (UPPA)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Biologie de la Reproduction, Environnement, Epigénétique & Développement (BREED), Université Paris-Saclay-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-École nationale vétérinaire d'Alfort (ENVA)-Université de Versailles Saint-Quentin-en-Yvelines (UVSQ), and J. L. received a doctoral fellowship from the China Scholarship Council (File No. 201506330063)

Subjects
Male, medicine.medical_specialty, [SDV]Life Sciences [q-bio], Medicine (miscellaneous), Biology, 03 medical and health sciences, 0302 clinical medicine, 5-carboxylcytosine, Internal medicine, Diet, Protein-Restricted, Dietary Carbohydrates, medicine, Animals, Dry matter, 5-hydroxymethylcytosine, 5-methylcytosine, 030304 developmental biology, Demethylation, 0303 health sciences, Nutrition and Dietetics, epigenetics, Methylation, Carbohydrate, biology.organism_classification, Diet, Trout, Phenotype, Endocrinology, DNA demethylation, 5-formylcytosine, Liver, Oncorhynchus mykiss, 030217 neurology & neurosurgery, DNA hypomethylation
Abstract

International audience; Abstract Background A high carbohydrate-low protein diet can induce hepatic global DNA hypomethylation in trout. The mechanisms remain unclear. Objective We aimed to investigate whether increase in dietary carbohydrates (dHC) or decrease in dietary proteins (dLP) can cause hepatic global DNA hypomethylation, and to explore the underlying mechanisms in trout. Methods Two feeding trials were conducted on juvenile males, both of which involved a 4-day fasting and 4-day refeeding protocol. In Trial 1, trout were fed either a high protein-no carbohydrate (HP-NC, protein 60% dry matter (DM), carbohydrates 0% DM) or a moderate protein-high carbohydrate (MP-HC, protein 40% DM, carbohydrates 30% DM) diet. In Trial 2, fish were fed either a moderate protein-no carbohydrate (MP-NC, protein 40% DM, carbohydrates 0% DM), a MP-HC (protein 40% DM, carbohydrates 30% DM), or a low protein-no carbohydrate (LP-NC, protein 20% DM, carbohydrates 0% DM) diet to separate the effects of dHC and dLP on the hepatic methylome. Global CmCGG methylation, DNA demethylation derivative levels, and mRNA expression of DNA (de)methylation-related genes were measured. Differences were tested by one-way ANOVA when data were normally distributed or by Kruskal-Wallis non-parametric test if not. Results In both trails, global CmCGG methylation levels remained unaffected, but the hepatic 5-mdC content decreased after refeeding (1–3%). The MP-HC group had 3.4-fold higher hepatic 5-hmdC and a similar 5-mdC level compared to the HP-NC group in Trial 1. Both MP-HC and LP-NC diets lowered the hepatic 5-mdC content (1–2%), but only the LP-NC group had a significantly lower 5-hmdC level (P

Published
2021

20. Analysis of the transcriptome of bovine endometrial cells isolated by laser micro-dissection (2): impacts of post-partum negative energy balance on stromal, glandular and luminal epithelial cells

Author

Patrice Humblot, Gilles Charpigny, Olivier Sandra, Mariam Raliou, Wiruntita Chankeaw, Göran Andersson, Claudia Bevilacqua, Yongzhi Guo, Christophe Richard, Theodoros Ntallaris, Sandra Lignier, Damien Plassard, Swedish University of Agricultural Sciences (SLU), Biologie de la Reproduction, Environnement, Epigénétique & Développement (BREED), Université Paris-Saclay-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-École nationale vétérinaire d'Alfort (ENVA)-Université de Versailles Saint-Quentin-en-Yvelines (UVSQ), Institut de Génétique et de Biologie Moléculaire et Cellulaire (IGBMC), Université de Strasbourg (UNISTRA)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Génétique Animale et Biologie Intégrative (GABI), Université Paris-Saclay-AgroParisTech-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Department of Animal Bredding and Genetics, FP7-KBBE-2012-6 Prolific and Grant agreement number 311776/Seventh Framework Programme, Charpigny, gilles, Collaborative Research: Sediment Nanomagnetism and Environmental Change: The Microbial Mineral Link - 2003-09-01 - 2007-08-31 - 0311776 - VALID, Rajamangala University of Technology Thanyaburi (RMUTT), École nationale vétérinaire - Alfort (ENVA)-Université de Versailles Saint-Quentin-en-Yvelines (UVSQ)-Université Paris-Saclay-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), AgroParisTech-Université Paris-Saclay-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), European Project: 0311776(2003), École nationale vétérinaire d'Alfort (ENVA)-Université de Versailles Saint-Quentin-en-Yvelines (UVSQ)-Université Paris-Saclay-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), and Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA)

Subjects
Cell type, Stromal cell, Negative energy balance, RNA-Seq, Biology, QH426-470, Transcriptome, Andrology, 03 medical and health sciences, Endometrium, Pregnancy, Animal and Dairy Science, Cell type-specific, Gene expression, Genetics, Animals, Humans, Lactation, Gene, [SDV.BDLR] Life Sciences [q-bio]/Reproductive Biology, 030304 developmental biology, Inflammation, 0303 health sciences, [SDV.BA.MVSA]Life Sciences [q-bio]/Animal biology/Veterinary medicine and animal Health, Dissection, Lasers, Postpartum Period, 0402 animal and dairy science, [SDV.BA.MVSA] Life Sciences [q-bio]/Animal biology/Veterinary medicine and animal Health, Embryo, Epithelial Cells, [SDV.BDLR]Life Sciences [q-bio]/Reproductive Biology, LCM, 04 agricultural and veterinary sciences, Acquired immune system, 040201 dairy & animal science, Cattle, Female, RNA-seq, Energy Metabolism, TP248.13-248.65, Research Article, Biotechnology
Abstract

Background In post-partum dairy cows, the energy needs to satisfy high milk production induces a status of more or less pronounced Negative Energy Balance (NEB). NEB associated with fat mobilization impairs reproductive function. In a companion paper, we described constitutive gene expression in the three main endometrial cell types (stromal, glandular and luminal epithelial cells) isolated by laser capture micro-dissection (LCM) showing the specificities of their transcriptomic profiles. This study investigates the specific impact of NEB on gene expression in these cells around 80 days after parturition at day 15 of the oestrus cycle and describes their specific response to NEB. Results Following the description of their constitutive expression, the transcriptome profiles obtained by RNA sequencing of the three cells types revealed that differences related to the severity of NEB altered mainly specific patterns of expression related to individual cell types. Number of differentially expressed genes between severe NEB (SNEB) and mild NEB (MNEB) cows was higher in ST than in LE and GE, respectively. SNEB was associated with differential expression of genes coding for proteins involved in metabolic processes and embryo-maternal interactions in ST. Under-expression of genes encoding proteins with functions related to cell structure was found in GE whereas genes encoding proteins participating in pro-inflammatory pathways were over-expressed. Genes associated to adaptive immunity were under-expressed in LE. Conclusion The severity of NEB after calving is associated with changes in gene expression around 80 days after parturition corresponding to the time of breeding. Specific alterations in GEs are associated with activation of pro-inflammatory mechanisms. Concomitantly, changes in the expression of genes encoding proteins involved in cell interactions and maternal recognition of pregnancy takes place in ST. The combination of these effects possibly altering the uterine environment and embryo maternal interactions may negatively influence the establishment of pregnancy.

Published
2021

21. Analysis of the transcriptome of bovine endometrial cells isolated by laser micro-dissection (1): specific signatures of stromal, glandular and luminal epithelial cells

Author

Christophe Richard, Olivier Sandra, Theodoros Ntallaris, Gilles Charpigny, Wiruntita Chankeaw, Yongzhi Guo, Sandra Lignier, Patrice Humblot, Göran Andersson, Mariam Raliou, Claudia Bevilacqua, Damien Plassard, Swedish University of Agricultural Sciences (SLU), Biologie de la Reproduction, Environnement, Epigénétique & Développement (BREED), École nationale vétérinaire d'Alfort (ENVA)-Université de Versailles Saint-Quentin-en-Yvelines (UVSQ)-Université Paris-Saclay-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Institut de Génétique et de Biologie Moléculaire et Cellulaire (IGBMC), Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA), Génétique Animale et Biologie Intégrative (GABI), Université Paris-Saclay-AgroParisTech-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Department of Animal Bredding and Genetics, 311776/FP7-KBBE-2016-6, PROLIFIC, Université Paris-Saclay-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-École nationale vétérinaire d'Alfort (ENVA)-Université de Versailles Saint-Quentin-en-Yvelines (UVSQ), Université de Strasbourg (UNISTRA)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Charpigny, gilles, Collaborative Research: Sediment Nanomagnetism and Environmental Change: The Microbial Mineral Link - 2003-09-01 - 2007-08-31 - 0311776 - VALID, École nationale vétérinaire - Alfort (ENVA)-Université de Versailles Saint-Quentin-en-Yvelines (UVSQ)-Université Paris-Saclay-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), AgroParisTech-Université Paris-Saclay-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), and European Project: 0311776(2003)

Subjects
Cell signaling, Stromal cell, Biology, QH426-470, Endometrium, Transcriptome, 03 medical and health sciences, 0302 clinical medicine, Pregnancy, Cell type-specific, medicine, Genetics, Animals, Embryo Implantation, Cell adhesion, [SDV.BDLR] Life Sciences [q-bio]/Reproductive Biology, 030304 developmental biology, Enzyme linked receptor protein signaling pathway, 0303 health sciences, 030219 obstetrics & reproductive medicine, Dissection, Lasers, Cow, Cilium movement, [SDV.BDLR]Life Sciences [q-bio]/Reproductive Biology, LCM, Epithelial Cells, Clinical Science, Cell biology, [SDV.AEN] Life Sciences [q-bio]/Food and Nutrition, medicine.anatomical_structure, Cattle, Female, RNA-seq, Stromal Cells, [SDV.AEN]Life Sciences [q-bio]/Food and Nutrition, TP248.13-248.65, Extracellular matrix organization, Biotechnology, Research Article
Abstract

Background A number of studies have examined mRNA expression profiles of bovine endometrium at estrus and around the peri-implantation period of pregnancy. However, to date, these studies have been performed on the whole endometrium which is a complex tissue. Consequently, the knowledge of cell-specific gene expression, when analysis performed with whole endometrium, is still weak and obviously limits the relevance of the results of gene expression studies. Thus, the aim of this study was to characterize specific transcriptome of the three main cell-types of the bovine endometrium at day-15 of the estrus cycle. Results In the RNA-Seq analysis, the number of expressed genes detected over 10 transcripts per million was 6622, 7814 and 8242 for LE, GE and ST respectively. ST expressed exclusively 1236 genes while only 551 transcripts were specific to the GE and 330 specific to LE. For ST, over-represented biological processes included many regulation processes and response to stimulus, cell communication and cell adhesion, extracellular matrix organization as well as developmental process. For GE, cilium organization, cilium movement, protein localization to cilium and microtubule-based process were the only four main biological processes enriched. For LE, over-represented biological processes were enzyme linked receptor protein signaling pathway, cell-substrate adhesion and circulatory system process. Conclusion The data show that each endometrial cell-type has a distinct molecular signature and provide a significantly improved overview on the biological process supported by specific cell-types. The most interesting result is that stromal cells express more genes than the two epithelial types and are associated with a greater number of pathways and ontology terms.

Published
2021

22. Transcription of rRNA in early mouse embryos promotes chromatin reorganization and expression of major satellite repeats

Author

Martine Chebrout, Maïmouna Coura Koné, Habib U. Jan, Marie Cournut, Martine Letheule, Renaud Fleurot, Tiphaine Aguirre-Lavin, Nathalie Peynot, Alice Jouneau, Nathalie Beaujean, Amélie Bonnet-Garnier, Biologie de la Reproduction, Environnement, Epigénétique & Développement (BREED), École nationale vétérinaire - Alfort (ENVA)-Université de Versailles Saint-Quentin-en-Yvelines (UVSQ)-Université Paris-Saclay-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Physiologie de la reproduction et des comportements [Nouzilly] (PRC), Institut Français du Cheval et de l'Equitation [Saumur] (IFCE)-Université de Tours (UT)-Centre National de la Recherche Scientifique (CNRS)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Unité Expérimentale de Physiologie Animale de l‘Orfrasiére (UE PAO), Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Institut cellule souche et cerveau / Stem Cell and Brain Research Institute (U1208 Inserm - UCBL1 / SBRI - USC 1361 INRAE), Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), and Bonnet-Garnier, Amelie

Subjects
Ribosomal DNA, Mouse, Transcription, Genetic, [SDV]Life Sciences [q-bio], Repeated sequences, rDNA transcription inhibition, Cell Biology, Embryo, Mammalian, DNA, Ribosomal, Chromatin, [SDV] Life Sciences [q-bio], Mice, Embryo, RNA, Ribosomal, Animals, Ribosomes, 3D-FISH
Abstract

During the first cell cycles of early development, the chromatin of the embryo is highly reprogrammed while the embryonic genome starts its own transcription. The spatial organization of the genome is an important process that contributes to regulating gene transcription in time and space. It has, however, been poorly studied in the context of early embryos. To study the cause-and-effect link between transcription and spatial organization in embryos, we focused on ribosomal genes, which are silent initially but start to be transcribed in 2-cell mouse embryos. We demonstrated that ribosomal sequences and early unprocessed rRNAs are spatially organized in a very particular manner between 2-cell and 16-cell stage. By using drugs that interfere with ribosomal DNA transcription, we showed that this organization – which is totally different in somatic cells – depends on an active transcription of ribosomal genes and induces a unique chromatin environment that favors transcription of major satellite sequences once the 4-cell stage has been reached.

Published
2021

23. Analysis of Polycerate Mutants Reveals the Evolutionary Co-option of HOXD1 for Horn Patterning in Bovidae

Author

Daniele Bigi, Fiona Menzi, Slim Ben Jemaa, Marie-Christine Deloche, Aurélien Capitan, Johannes A. Lenstra, Marina Naval-Sanchez, Ivica Medugorac, Nathalie Hirter, Gwenola Tosser-Klopp, Diane Esquerre, Coralie M. Reich, Julia M. Paris, Rose-Marie Arbogast, Amandine Blin, Abdelhak Boukadiri, Aurélie Hintermann, Julie Rivière, Denis Duboule, Raphaël Cornette, Cécile Donnadieu, Marie-Dominique Wandhammer, Gjoko Bunevski, Louisa Gidney, Michael Stache, Isabelle Palhiere, Renate Schafberg, James Kijas, Claude Guintard, Joséphine Lesur, Jozsef Zakany, Rachel Rupp, Noelle E. Cockett, John Hedges, Ashleigh Haruda, Philippe Bardou, Olivier Putelat, Tracy Hadfield, Alain Pinton, Ockert Greyvenstein, Aurélie Allais-Bonnet, Este Van Marle-Koster, Eric Pailhoux, Coralie Danchin-Burge, David G. Riley, Cécile Grohs, Benjamin J. Hayes, Cord Drögemüller, Allice, Biologie de la Reproduction, Environnement, Epigénétique & Développement (BREED), Université Paris-Saclay-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-École nationale vétérinaire d'Alfort (ENVA)-Université de Versailles Saint-Quentin-en-Yvelines (UVSQ), Institut de Systématique, Evolution, Biodiversité (ISYEB ), Muséum national d'Histoire naturelle (MNHN)-École pratique des hautes études (EPHE), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS)-Université des Antilles (UA), Génétique Physiologie et Systèmes d'Elevage (GenPhySE ), Ecole Nationale Vétérinaire de Toulouse (ENVT), Institut National Polytechnique (Toulouse) (Toulouse INP), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Institut National Polytechnique (Toulouse) (Toulouse INP), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-École nationale supérieure agronomique de Toulouse [ENSAT]-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Commonwealth Scientific and Industrial Research Organisation [Canberra] (CSIRO), École nationale supérieure agronomique de Toulouse [ENSAT]-Institut National de la Recherche Agronomique (INRA)-Ecole Nationale Vétérinaire de Toulouse (ENVT), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées, Martin-Luther-University Halle-Wittenberg, Génétique Animale et Biologie Intégrative (GABI), Université Paris-Saclay-AgroParisTech-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), University of Geneva [Switzerland], University of Bologna, Ludwig Maximilian University [Munich] (LMU), Archéologie d'Alsace, Archéologie et histoire ancienne : Méditerranée - Europe (ARCHIMEDE), Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Ministère de la Culture et de la Communication (MCC)-Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA), Texas A&M University [College Station], Utah State University (USU), Université de Carthage - University of Carthage, Ss. Cyril and Methodius University in Skopje, University of Bern, Manx Text, Manx Loaghtan Sheep Breeders (MLSBG), Utrecht University [Utrecht], Rent a Peasant, Archéozoologie, archéobotanique : sociétés, pratiques et environnements (AASPE), Muséum national d'Histoire naturelle (MNHN)-Centre National de la Recherche Scientifique (CNRS), Musée Zoologique de Strasbourg, Ecole Nationale Vétérinaire, Agroalimentaire et de l'alimentation Nantes-Atlantique (ONIRIS), Acquisition et Analyse de Données pour l'Histoire naturelle (2AD), MICrobiologie de l'ALImentation au Service de la Santé (MICALIS), AgroParisTech-Université Paris-Saclay-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Génome et Transcriptome - Plateforme Génomique ( GeT-PlaGe), Plateforme Génome & Transcriptome (GET), Génopole Toulouse Midi-Pyrénées [Auzeville] (GENOTOUL), Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Ecole Nationale Vétérinaire de Toulouse (ENVT), Université Fédérale Toulouse Midi-Pyrénées-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), Université Fédérale Toulouse Midi-Pyrénées-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Génopole Toulouse Midi-Pyrénées [Auzeville] (GENOTOUL), Université Fédérale Toulouse Midi-Pyrénées-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Institut de l'élevage (IDELE), Agriculture Victoria (AgriBio), Tshwane University of Technology [Pretoria] (TUT), Queensland Alliance for Agriculture and Food Innovation (QAAFI), University of Queensland [Brisbane], CSIRO Agriculture and Food (CSIRO), Ecole Polytechnique Fédérale de Lausanne (EPFL), Collège de France (CdF (institution)), Allais-Bonnet, Aurélie, Hintermann, Aurélie, Deloche, Marie-Christine, Cornette, Raphaël, Bardou, Philippe, Naval-Sanchez, Marina, Pinton, Alain, Haruda, Ashleigh, Grohs, Cécile, Zakany, Jozsef, Bigi, Daniele, Medugorac, Ivica, Putelat, Olivier, Greyvenstein, Ockert, Hadfield, Tracy, Jemaa, Slim Ben, Bunevski, Gjoko, Menzi, Fiona, Hirter, Nathalie, Paris, Julia M, Hedges, John, Palhiere, Isabelle, Rupp, Rachel, Lenstra, Johannes A, Gidney, Louisa, Lesur, Joséphine, Schafberg, Renate, Stache, Michael, Wandhammer, Marie-Dominique, Arbogast, Rose-Marie, Guintard, Claude, Blin, Amandine, Boukadiri, Abdelhak, Rivière, Julie, Esquerré, Diane, Donnadieu, Cécile, Danchin-Burge, Coralie, Reich, Coralie M, Riley, David G, Marle-Koster, Este van, Cockett, Noelle, Hayes, Benjamin J, Drögemüller, Cord, Kijas, Jame, Pailhoux, Eric, Tosser-Klopp, Gwenola, Duboule, Deni, Capitan, Aurélien, École nationale vétérinaire d'Alfort (ENVA)-Université de Versailles Saint-Quentin-en-Yvelines (UVSQ)-Université Paris-Saclay-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Ministère de la Culture et de la Communication (MCC)-Centre National de la Recherche Scientifique (CNRS), Centre National de la Recherche Scientifique (CNRS)-Muséum national d'Histoire naturelle (MNHN), Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Université Toulouse III - Paul Sabatier (UT3), École nationale vétérinaire - Alfort (ENVA)-Université de Versailles Saint-Quentin-en-Yvelines (UVSQ)-Université Paris-Saclay-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Institut National de la Recherche Agronomique (INRA)-Ecole Nationale Vétérinaire de Toulouse (ENVT), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-École nationale supérieure agronomique de Toulouse [ENSAT], Université de Genève = University of Geneva (UNIGE), University of Bologna/Università di Bologna, Ss. Cyril and Methodius University in Skopje (UKIM), École nationale vétérinaire, agroalimentaire et de l'alimentation Nantes-Atlantique (ONIRIS), and Collège de France - Chaire internationale Évolution des génomes et développement

Subjects
0106 biological sciences, Male, co-option, translocation, AcademicSubjects/SCI01180, regulatory mutation, 01 natural sciences, Hox genes, Hox gene, genes, 610 Medicine & health, Bilateria, time, Horns, 0303 health sciences, biology, Ecology, 630 Agriculture, Goats, [SDV.BDD.EO] Life Sciences [q-bio]/Development Biology/Embryology and Organogenesis, Gene Expression Regulation, Developmental, alignment, Morphogenetic field, Biological Evolution, [SDV.BDD.MOR] Life Sciences [q-bio]/Development Biology/Morphogenesis, 590 Animals (Zoology), Haploinsufficiency, transcription, Biometry, Evolution, growth, Locus (genetics), Mice, Transgenic, [SDV.GEN.GA] Life Sciences [q-bio]/Genetics/Animal genetics, 010603 evolutionary biology, 03 medical and health sciences, goat and sheep genomics, Behavior and Systematics, framework, Genetics, Animals, Allele, Gene, Molecular Biology, Discoveries, Ecology, Evolution, Behavior and Systematics, 030304 developmental biology, Homeodomain Proteins, locus, Sheep, Horn (anatomy), AcademicSubjects/SCI01130, [SDV.BDD.MOR]Life Sciences [q-bio]/Development Biology/Morphogenesis, sequence, biology.organism_classification, [SDV.GEN.GA]Life Sciences [q-bio]/Genetics/Animal genetics, [SDV.BDD.EO]Life Sciences [q-bio]/Development Biology/Embryology and Organogenesis, Evolutionary biology, Mutation, genome-wide association, 570 Life sciences, goat and sheep genomic
Abstract

In the course of evolution, pecorans (i.e., higher ruminants) developed a remarkable diversity of osseous cranial appendages, collectively referred to as “headgear,” which likely share the same origin and genetic basis. However, the nature and function of the genetic determinants underlying their number and position remain elusive. Jacob and other rare populations of sheep and goats are characterized by polyceraty, the presence of more than two horns. Here, we characterize distinct POLYCERATE alleles in each species, both associated with defective HOXD1 function. We show that haploinsufficiency at this locus results in the splitting of horn bud primordia, likely following the abnormal extension of an initial morphogenetic field. These results highlight the key role played by this gene in headgear patterning and illustrate the evolutionary co-option of a gene involved in the early development of bilateria to properly fix the position and number of these distinctive organs of Bovidae.

Published
2021

24. Metabolomic differences in blastocoel and uterine fluids collected in vivo by ultrasound biomicroscopy on rabbit embryos

Author

Michele Dahirel, Gilles Renault, Nathalie Daniel, Eve Mourier, Carmen Marchiol, Julie Gatien, Lydie Nadal-Desbarats, Véronique Duranthon, Christophe Richard, Franck Lager, Pascale Chavatte-Palmer, Sophie Calderari, Biologie de la Reproduction, Environnement, Epigénétique & Développement (BREED), Université Paris-Saclay-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-École nationale vétérinaire d'Alfort (ENVA)-Université de Versailles Saint-Quentin-en-Yvelines (UVSQ), Plateforme MIMA2, Institut National de la Recherche Agronomique (INRA), Institut Cochin (IC UM3 (UMR 8104 / U1016)), Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université de Paris (UP), Allice, Imagerie et cerveau (iBrain - Inserm U1253 - UNIV Tours ), Université de Tours-Institut National de la Santé et de la Recherche Médicale (INSERM), Plateforme Scientifique et Technique 'Analyses des Systèmes Biologiques' (PST-ASB), Université Francois Rabelais [Tours], École nationale vétérinaire d'Alfort (ENVA)-Université de Versailles Saint-Quentin-en-Yvelines (UVSQ)-Université Paris-Saclay-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), and Université de Tours (UT)-Institut National de la Santé et de la Recherche Médicale (INSERM)

Subjects
0301 basic medicine, metabolite, Microscopy, Acoustic, Uterus, Ultrasound biomicroscopy, rabbit, Biology, Andrology, 03 medical and health sciences, 0302 clinical medicine, Metabolomics, Pregnancy, In vivo, medicine, Animals, [SDV.BDD]Life Sciences [q-bio]/Development Biology, blastocoel, 030219 obstetrics & reproductive medicine, [SDV.BA]Life Sciences [q-bio]/Animal biology, Embryogenesis, Blastocoel, Rabbit (nuclear engineering), Embryo, [SDV.BDLR]Life Sciences [q-bio]/Reproductive Biology, Cell Biology, General Medicine, pre-implantation embryo, Embryo, Mammalian, Body Fluids, Blastocyst, 030104 developmental biology, medicine.anatomical_structure, Reproductive Medicine, uterine fluid, Metabolome, Female, Rabbits, ultrasound biomicroscopy
Abstract

The success of embryo development and implantation depends in part on the environment in which the embryo evolves. However, the composition of the uterine fluid surrounding the embryo in the peri-implantation period remains poorly studied. In this work, we aimed to develop a new strategy to visualize, collect, and analyze both blastocoelic liquid and juxta-embryonic uterine fluid from in vivo peri-implantation rabbit embryos. Using high-resolution ultrasound biomicroscopy, embryos were observed as fluid-filled anechoic vesicles, some of which were surrounded by a thin layer of uterine fluid. Ultrasound-guided puncture and aspiration of both the blastocoelic fluid contained in the embryo and the uterine fluid in the vicinity of the embryo were performed. Using nuclear magnetic resonance spectroscopy, altogether 24 metabolites were identified and quantified, of which 21 were detected in both fluids with a higher concentration in the uterus compared to the blastocoel. In contrast, pyruvate was detected at a higher concentration in blastocoelic compared to uterine fluid. Two acidic amino acids, glutamate and aspartate, were not detected in uterine fluid in contrast to blastocoelic fluid, suggesting a local regulation of uterine fluid composition. To our knowledge, this is the first report of simultaneous analysis of blastocoelic and uterine fluids collected in vivo at the time of implantation in mammals, shedding new insight for understanding the relationship between the embryo and its local environment at this critical period of development.

Published
2021

25. Human apolipoprotein C1 transgenesis reduces atherogenesis in hypercholesterolemic rabbits

Author

Erwana Harscoët, Naig Le Guern, Stéphanie Lemaire, Valérie Deckert, Bruno Da Silva, Geneviève Jolivet, Thomas Gautier, Lil Proukhnitzky, Marion Xolin, Amandine Bataille, Laurent Lagrost, Charlène Magnani, Virginie Aires, Louis-Marie Houdebine, David Masson, Guillaume Maquart, Danish Patoli, UFR des Sciences de Santé (Université de Bourgogne), Université de Bourgogne (UB), Centre Hospitalier Universitaire de Dijon - Hôpital François Mitterrand (CHU Dijon), Biologie de la Reproduction, Environnement, Epigénétique & Développement (BREED), École nationale vétérinaire - Alfort (ENVA)-Université de Versailles Saint-Quentin-en-Yvelines (UVSQ)-Université Paris-Saclay-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Laboratoire de Biologie du Développement [IBPS] (LBD), Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS)-Institut de Biologie Paris Seine (IBPS), Institut National de la Santé et de la Recherche Médicale (INSERM)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), This work was supported by grants from the University of Bourgogne Franche-Comté, the Institut National de la Santé et de la Recherche Médicale (INSERM), the Fondation de France [Project Cardio 2013 R13057MM-RAF13002MMA to TG], ANR grant ATHEROLIP, the European Union program FEDER, and by a French Government grant managed by the French National Research Agency under the program ‘Investissements d’Avenir’ [reference ANR-11-LABX-0021 (Lipstic Labex)]., ANR-11-LABX-0021,Lipstic,Lipoprotéines et santé : prévention et traitement des maladies inflammatoires non vasculaires et du cancer(2011), ANR-06-PHYS-0002,ATHEROLIP,Identification et caractérisation des propriétés proathérogènes des protéines plasmatiques de transfert des lipides (CETP et PLTP).(2006), European Project: FEDER, Jolivet, Geneviève, Laboratoires d'excellence - Lipoprotéines et santé : prévention et traitement des maladies inflammatoires non vasculaires et du cancer - - Lipstic2011 - ANR-11-LABX-0021 - LABX - VALID, Physiopathologie des maladies humaines (Physio) - Identification et caractérisation des propriétés proathérogènes des protéines plasmatiques de transfert des lipides (CETP et PLTP). - - ATHEROLIP2006 - ANR-06-PHYS-0002 - PHYSIO - VALID, Fonds Européens de Développement Régional - FEDER - INCOMING, Université Paris-Saclay-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-École nationale vétérinaire d'Alfort (ENVA)-Université de Versailles Saint-Quentin-en-Yvelines (UVSQ), Laboratoire de Biologie du Développement [Paris] (LBD), and ANR-11-LABX-0021,Lipstic,Lipoprotéines et santé : prévention et traitement des maladies inflammatoires non vasculaires et du(2011)

Subjects
0301 basic medicine, medicine.medical_specialty, Very low-density lipoprotein, Apolipoprotein B, HDL, Transgene, Apolipoprotein C1, Endogeny, Rabbit, 030204 cardiovascular system & hematology, Transgenic, 03 medical and health sciences, chemistry.chemical_compound, Mice, 0302 clinical medicine, [SDV.MHEP.CSC]Life Sciences [q-bio]/Human health and pathology/Cardiology and cardiovascular system, Internal medicine, [SDV.BBM.GTP]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Genomics [q-bio.GN], CETP, medicine, Animals, Humans, 2. Zero hunger, Apolipoprotein C-I, biology, Triglyceride, Cholesterol, Cholesterol, HDL, Gene Transfer Techniques, Atherosclerosis, Cholesterol Ester Transfer Proteins, [SDV.MHEP.CSC] Life Sciences [q-bio]/Human health and pathology/Cardiology and cardiovascular system, carbohydrates (lipids), Transgenesis, 030104 developmental biology, Endocrinology, chemistry, biology.protein, [SDV.BBM.GTP] Life Sciences [q-bio]/Biochemistry, Molecular Biology/Genomics [q-bio.GN], lipids (amino acids, peptides, and proteins), Rabbits, Cardiology and Cardiovascular Medicine
Abstract

International audience; Background and aims: Apolipoprotein (apo) C1 is a 6.6 kDa protein associated with HDL and VLDL. ApoC1 alters triglyceride clearance, and it also favors cholesterol accumulation in HDL, especially by inhibiting CETP in human plasma. Apart from studies in mice, which lack CETP, the impact of apoC1 on atherosclerosis in animal models expressing CETP, like in humans, is not known. This study aimed at determining the net effect of human apoC1 on atherosclerosis in rabbits, a species with naturally high CETP activity but with endogenous apoC1 without CETP inhibitory potential.Methods: Rabbits expressing a human apoC1 transgene (HuApoC1Tg) were generated and displayed significant amounts of human apoC1 in plasma.Results: After cholesterol feeding, atherosclerosis lesions were significantly less extensive (-22%, p < 0.05) and HDL displayed a reduced ability to serve as CETP substrates (-25%, p < 0.05) in HuApoC1Tg rabbits than in WT littermates. It was associated with rises in plasma HDL cholesterol level and PON-1 activity, and a decrease in the plasma level of the lipid oxidation markers 12(S)-HODE and 8(S)HETE. In chow-fed animals, the level of HDL-cholesterol was also significantly higher in HuApoC1Tg than in WT animals (0.83 ± 0.11 versus 0.73 ± 0.11 mmol/L, respectively, p < 0.05), and it was associated with significantly lower CETP activity (cholesteryl ester transfer rate, -10%, p < 0.05; specific CETP activity, -14%, p < 0.05).Conclusions: Constitutive expression of fully functional human apoC1 in transgenic rabbit attenuates atherosclerosis. It was found to relate, at least in part, to the inhibition of plasma CETP activity and to alterations in plasma HDL.

Published
2021

26. Pressurized intra-peritoneal aerosol chemotherapy (PIPAC): increased intraperitoneal pressure does not affect distribution patterns but leads to deeper penetration depth of doxorubicin in a sheep model

Author

Myriam Mimouni, Pierre Adenot, Julien Cohen, François Vialard, Arnaud Fauconnier, Thomas Lilin, Christophe Richard, Valérie Gelin, Pascale Chavatte-Palmer, Martine Letheule, Benoit Lecuelle, Cyrille Huchon, Michèle Dahirel, Olivier Sandra, Anne Tarrade, Biologie de la Reproduction, Environnement, Epigénétique & Développement (BREED), École nationale vétérinaire d'Alfort (ENVA)-Université de Versailles Saint-Quentin-en-Yvelines (UVSQ)-Université Paris-Saclay-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Service de gynécologie et obstétrique [CHI Poissy-Saint Germain], CHI Poissy-Saint-Germain, Centre de recherche biomédicale (CRBM), École nationale vétérinaire d'Alfort (ENVA)-Université Paris-Est (UPE), Risques cliniques et sécurité en santé des femmes et en santé périnatale (RISCQ), Université de Versailles Saint-Quentin-en-Yvelines (UVSQ), Hôpital Lariboisière-Fernand-Widal [APHP], and Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)

Subjects
Cancer Research, medicine.medical_treatment, PIPAC, Ovary, [SDV.CAN]Life Sciences [q-bio]/Cancer, Andrology, Caecum, 03 medical and health sciences, chemistry.chemical_compound, Peritoneal cavity, 0302 clinical medicine, Drug Delivery Systems, Peritoneum, Genetics, medicine, Pressure, Animals, Doxorubicin, Tissue Distribution, DAPI, Intraperitoneum pressure, Cecum, RC254-282, Peritoneal Neoplasms, Aerosols, Cell Nucleus, Chemotherapy, Antibiotics, Antineoplastic, Sheep, biology, Neoplasms. Tumors. Oncology. Including cancer and carcinogens, Penetration (firestop), [SDV.SP]Life Sciences [q-bio]/Pharmaceutical sciences, biology.organism_classification, medicine.anatomical_structure, Oncology, chemistry, 030220 oncology & carcinogenesis, 030211 gastroenterology & hepatology, Female, Omentum, Peritoneal carcinomatosis, medicine.drug, Research Article
Abstract

Background Pressurized Intra-Peritoneal Aerosol Chemotherapy (PIPAC) is an innovative treatment against peritoneal carcinomatosis. Doxorubicin is a common intra-venous chemotherapy used for peritoneal carcinomatosis and for PIPAC. This study evaluated the impact of increased PIPAC intraperitoneal pressure on the distribution and cell penetration of doxorubicin in a sheep model. Methods Doxorubicin was aerosolized using PIPAC into the peritoneal cavity of 6 ewes (pre-alpes breed): N = 3 with 12 mmHg intraperitoneal pressure (“group 12”) and N = 3 with 20 mmHg (“group 20”). Samples from peritoneum (N = 6), ovarian (N = 1), omentum (N = 1) and caecum (N = 1) were collected for each ewe. The number of doxorubicin positive cells was determined using the ratio between doxorubicine fluorescence-positive cell nuclei (DOXO+) over total number of DAPI positive cell nuclei (DAPI+). Penetration depth (μm) was defined as the distance between the luminal surface and the location of the deepest DOXO+ nuclei over the total number of cell nuclei that were stained with DAPI. Penetration depth (μm) was defined as the distance between the luminal surface and the location of the deepest DOXO+ nuclei. Results DOXO+ nuclei were identified in 87% of samples. All omental samples, directly localized in front of the nebulizer head, had 100% DOXO+ nuclei whereas very few nuclei were DOXO+ for caecum. Distribution patterns were not different between the two groups but penetration depth in ovary and caecum samples was significantly deeper in group 20. Conclusions This study showed that applying a higher intra-peritoneal pressure during PIPAC treatment leads to a deeper penetration of doxorubicin in ovarian and caecum but does not affect distribution patterns.

Published
2021

27. Nutrition of Broodmares

Author

Carolyn J. Hammer, Morgane Robles, Burt Staniar, Pascale Chavatte-Palmer, Institut National de la Recherche Scientifique [Québec] (INRS), Biologie de la Reproduction, Environnement, Epigénétique & Développement (BREED), École nationale vétérinaire d'Alfort (ENVA)-Université de Versailles Saint-Quentin-en-Yvelines (UVSQ)-Université Paris-Saclay-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), École nationale vétérinaire d'Alfort (ENVA), North Dakota State University (NDSU), Pennsylvania State University (Penn State), and Penn State System

Subjects
Nutritional Supplementation, 040301 veterinary sciences, animal diseases, Context (language use), Disease, 0403 veterinary science, Pregnancy, biology.animal, Environmental health, Animals, Medicine, Horses, Nutrition, 2. Zero hunger, [SDV.BA.MVSA]Life Sciences [q-bio]/Animal biology/Veterinary medicine and animal Health, biology, Equine, business.industry, 0402 animal and dairy science, Maternal Nutritional Physiological Phenomena, 04 agricultural and veterinary sciences, Animal Feed, 040201 dairy & animal science, Developmental origins of health and diseases (DOHaD), Diet, Foal, Diet quality, Animal Nutritional Physiological Phenomena, Female, Critical periods, business, Gestation, Broodmare
Abstract

International audience; Forage availability should cover most needs for mares bred during spring and summer. Out-of-season breeding, lack of access to pasture, or good quality forage calls for nutritional supplementation. Current evaluations of broodmare needs are based on fetoplacental tissue requirements, but do not consider endocrine changes or that the maternal diet quality affects long-term foal health. This article reviews pregnant mares' current nutritional recommendations. Secondly, fetoplacental developmental stages during gestation are outlined, defining critical periods in the context of the developmental origins of health and disease. Last, examples of how maternal nutrition affects long-term foal health are presented.

Published
2021

28. Amino acids activate mTORC1 to release roe deer embryos from decelerated proliferation during diapause

Author

van der Weijden, Vera A, Bick, Jochen T, Bauersachs, Stefan, Rüegg, Anna B, Hildebrandt, Thomas B, Goeritz, Frank, Jewgenow, Katarina, Giesbertz, Pieter, Daniel, Hannelore, Derisoud, Emilie, Chavatte-Palmer, Pascale, Bruckmaier, Rupert M, Drews, Barbara, Ulbrich, Susanne E, University of Zurich, Ulbrich, Susanne E, Animal Physiology, Institute of Agricultural sciences, ETH, Eidgenössische Technische Hochschule - Swiss Federal Institute of Technology [Zürich] (ETH Zürich), Universität Zürich [Zürich] = University of Zurich (UZH), Leibniz Institute for Zoo and Wildlife Research (IZW), Leibniz Association, Technische Universität München [München] (TUM), Biologie de la Reproduction, Environnement, Epigénétique & Développement (BREED), École nationale vétérinaire d'Alfort (ENVA)-Université de Versailles Saint-Quentin-en-Yvelines (UVSQ)-Université Paris-Saclay-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Institute of Veterinary Physiology, Vetsuisse Faculty, and Zurich Center for Integrative Human Physiology (ZIHP), Swiss National Science Foundation (31003A_159734 and 310030_185026), Technische Universität Munchen - Université Technique de Munich [Munich, Allemagne] (TUM), and Université Paris-Saclay-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-École nationale vétérinaire d'Alfort (ENVA)-Université de Versailles Saint-Quentin-en-Yvelines (UVSQ)

Subjects
Physiology, 10077 Institute of Veterinary Anatomy, gamete, embryo, Embryonic Development, Mechanistic Target of Rapamycin Complex 1, embryonic diapause, gonad, [SDV.BDLR.RS]Life Sciences [q-bio]/Reproductive Biology/Sexual reproduction, Pregnancy, Animals, roe deer, Amino Acids, European roe deer (Capreolus capreolus), Cell Proliferation, 1000 Multidisciplinary, [SDV.BA.MVSA]Life Sciences [q-bio]/Animal biology/Veterinary medicine and animal Health, 630 Agriculture, embryo development, Deer, Gene Expression Profiling, Uterus, Biological Sciences, Embryo, Mammalian, Diapause, Blastocyst, Cellular Microenvironment, 570 Life sciences, biology, Female
Abstract

Significance In mammals, embryo development can halt at the hatched blastocyst stage. Uniquely, proliferation of diapausing embryonic roe deer cells decelerates to a doubling time of 2 to 3 wk over a period of 4 mo. We highlight nutrient sensing as an important factor regulating embryonic developmental pace. The resumption of embryo development is characterized by an increase in uterine fluid mTORC1-activating amino acids, embryonic mTORC1 activity, and expression of metabolism and cell cycle genes. We propose selective mTORC1 inhibition via reduced estrogen signaling and high let-7 levels as mechanisms for slow cell cycle progression. We hypothesize that it is the lack of embryonic mTORC2 inhibition during embryonic diapause in the roe deer that enables the continuous decelerated rate of proliferation., Embryonic diapause in mammals leads to a reversible developmental arrest. While completely halted in many species, European roe deer (Capreolus capreolus) embryos display a continuous deceleration of proliferation. During a 4-mo period, the cell doubling time is 2 to 3 wk. During this period, the preimplantation blastocyst reaches a diameter of 4 mm, after which it resumes a fast developmental pace to subsequently implant. The mechanisms regulating this notable deceleration and reacceleration upon developmental resumption are unclear. We propose that amino acids of maternal origin drive the embryonic developmental pace. A pronounced change in the abundance of uterine fluid mTORC1-activating amino acids coincided with an increase in embryonic mTORC1 activity prior to the resumption of development. Concurrently, genes related to the glycolytic and phosphate pentose pathway, the TCA cycle, and one carbon metabolism were up-regulated. Furthermore, the uterine luminal epithelial transcriptome indicated increased estradiol-17β signaling, which likely regulates the endometrial secretions adapting to the embryonic needs. While mTORC1 was predicted to be inactive during diapause, the residual embryonic mTORC2 activity may indicate its involvement in maintaining the low yet continuous proliferation rate during diapause. Collectively, we emphasize the role of nutrient signaling in preimplantation embryo development. We propose selective mTORC1 inhibition via uterine catecholestrogens and let-7 as a mechanism regulating slow stem cell cycle progression.

Published
2021

29. Analysis of blood parameters and molecular endometrial markers during early reperfusion in two ovine models of uterus transplantation

Author

Barbara Hersant, Valérie Gelin, Laurent Galio, Mariam Raliou, François Vialard, Olivier Dubois, Anne Couturier-Tarrade, Corinne Giraud-Delville, Angéline Favre-Inhofer, Gilles Charpigny, Olivier Sandra, Nathalie Cornet, Jean-Marc Ayoubi, Romain Bosc, Aurélie Revaux, Marie Carbonnel, Pascale Chavatte-Palmer, Raphaël Coscas, Christophe Richard, Biologie de la Reproduction, Environnement, Epigénétique & Développement (BREED), École nationale vétérinaire - Alfort (ENVA)-Université de Versailles Saint-Quentin-en-Yvelines (UVSQ)-Université Paris-Saclay-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Hôpital Foch [Suresnes], Service de chirurgie plastique et reconstructive [Mondor], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Hôpital Henri Mondor-Université Paris-Est Créteil Val-de-Marne - Paris 12 (UPEC UP12), Hôpital Ambroise Paré [AP-HP], Centre de recherche en épidémiologie et santé des populations (CESP), Université de Versailles Saint-Quentin-en-Yvelines (UVSQ)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Hôpital Paul Brousse-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Paris-Saclay, The authors received funding (financial and material support) from the Foch Foundation. Angéline Favre-Inhofer and Nathalie Cornet were paid a stipend by the Foch Foundation. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript., Université Paris-Saclay-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-École nationale vétérinaire d'Alfort (ENVA)-Université de Versailles Saint-Quentin-en-Yvelines (UVSQ), and Chavatte-Palmer, Pascale

Subjects
[SDV.MHEP.CHI] Life Sciences [q-bio]/Human health and pathology/Surgery, Physiology, Uterus, 030230 surgery, Endometrium, Vascular Medicine, 0302 clinical medicine, Ischemia, Medicine and Health Sciences, Immune Response, 030219 obstetrics & reproductive medicine, Multidisciplinary, Anastomosis, Surgical, Arteries, Dissection, medicine.anatomical_structure, Reperfusion Injury, Gestation, Medicine, Female, Anatomy, Research Article, medicine.medical_specialty, Histology, Science, Immunology, Urology, [SDV.MHEP.CHI]Life Sciences [q-bio]/Human health and pathology/Surgery, Anastomosis, [SDV.MHEP.GEO]Life Sciences [q-bio]/Human health and pathology/Gynecology and obstetrics, [SDV.BDLR.RS]Life Sciences [q-bio]/Reproductive Biology/Sexual reproduction, Veins, 03 medical and health sciences, Signs and Symptoms, Uterus transplantation, medicine, Animals, Vein, Inflammation, Sheep, business.industry, Reproductive System, Biology and Life Sciences, medicine.disease, [SDV.MHEP.GEO] Life Sciences [q-bio]/Human health and pathology/Gynecology and obstetrics, Reperfusion, Cardiovascular Anatomy, Blood Vessels, Clinical Medicine, business, Physiological Processes, Transcriptome
Abstract

The dissection of the veins is the trickiest step of Uterine transplantation (UTx). Performing the anastomosis of a single uterine vein could bring a therapeutic benefit and simplification of surgery and serve for managing unilateral venous thromboses. The objectives of this project were to evaluate the expression of early markers of ischemia-reperfusion and to compare findings following one or two vein anastomoses. Orthotopic uterine auto-transplantations were performed on an ovine model with anastomosis of either two (group 1) or one utero-ovarian veins (group 2). Blood gases, histology and ischemia- reperfusion markers transcripts (PTGS2, IL6, IL8, SOD2, C3, BAX/BCL2 and TLR4) were analyzed as well as PTGS2 protein expression using Western Blot and fluorescence immunolocalization on endometrial biopsies after 3h of reperfusion. Ten ewes were included in the experimentation, 4 were in group1, 3 in group 2, the others being sham operated controls. No significant differences were observed between the two phenotypes. Based on these results, the anastomosis of one single uterine vein appears to be an approach consistent with short-term graft survival. Further experiments will be needed to confirm the reliability of this approach, especially the long-term follow-up of the uterine graft including its ability to support gestation to term.

Published
2021

30. Progress and challenges in developing organoids in farm animal species for the study of reproduction and their applications to reproductive biotechnologies

Author

Bourdon, Guillaume, Cadoret, Véronique, Charpigny, Gilles, Couturier-Tarrade, Anne, Dalbies-Tran, Rozenn, Flores, Maria-Jose, Froment, Pascal, Raliou, Mariam, Reynaud, Karine, Saint-Dizier, Marie, Jouneau, Alice, Physiologie de la reproduction et des comportements [Nouzilly] (PRC), Institut Français du Cheval et de l'Equitation [Saumur]-Université de Tours (UT)-Centre National de la Recherche Scientifique (CNRS)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Biologie de la Reproduction, Environnement, Epigénétique & Développement (BREED), École nationale vétérinaire d'Alfort (ENVA)-Université de Versailles Saint-Quentin-en-Yvelines (UVSQ)-Université Paris-Saclay-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), and Institut Français du Cheval et de l'Equitation [Saumur]-Université de Tours-Centre National de la Recherche Scientifique (CNRS)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)

Subjects
Follicle, Organoid, lcsh:Veterinary medicine, [SDV.BIO]Life Sciences [q-bio]/Biotechnology, Reproduction, Placenta, Ovary, Uterus, Cell Culture Techniques, [SDV.SA.ZOO]Life Sciences [q-bio]/Agricultural sciences/Zootechny, Review, Oviduct, [SDV.BDLR.RS]Life Sciences [q-bio]/Reproductive Biology/Sexual reproduction, Organoids, Embryoid, Endometrium, Reproductive Techniques, Animals, Domestic, Testis, lcsh:SF600-1100, Animals
Abstract

International audience; Within the past decades, major progress has been accomplished in isolating germ/stem/pluripotent cells, in refining culture medium and conditions and in establishing 3-dimensional culture systems, towards developing organoids for organs involved in reproduction in mice and to some extent in humans. Haploid male germ cells were generated in vitro from primordial germ cells. So were oocytes, with additional support from ovarian cells and subsequent follicle culture. Going on with the female reproductive tract, spherical oviduct organoids were obtained from adult stem/progenitor cells. Multicellular endometrial structures mimicking functional uterine glands were derived from endometrial cells. Trophoblastic stem cells were induced to form 3-dimensional syncytial-like structures and exhibited invasive properties, a crucial point for placentation. Finally, considering the embryo itself, pluripotent embryonic cells together with additional extra-embryonic cells, could self-organize into a blastoid, and eventually into a post-implantation-like embryo. Most of these accomplishments have yet to be reached in farm animals, but much effort is devoted towards this goal. Here, we review the progress and discuss the specific challenges of developing organoids for the study of reproductive biology in these species. We consider the use of such organoids in basic research to delineate the physiological mechanisms involved at each step of the reproductive process, or to understand how they are altered by environmental factors relevant to animal breeding. We evaluate their potential in reproduction of animals with a high genetic value, from a breeding point of view or in the context of preserving local breeds with limited headcounts.

Published
2021

31. Challenging the Ex Vivo Lung Perfusion Procedure With Continuous Dialysis in a Pig Model

Author

Antoine Roux, Isabelle Schwartz-Cornil, Matthieu Glorion, Edouard Sage, Fany Blanc, Morgan Le Guen, Jean-Jacques Leplat, Didier Journois, Céline Urien, Luc Jouneau, Julien De Wolf, Jérôme Estephan, Christophe Richard, Hôpital Foch [Suresnes], Virologie et Immunologie Moléculaires (VIM (UR 0892)), Université de Versailles Saint-Quentin-en-Yvelines (UVSQ)-Université Paris-Saclay-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Génétique Animale et Biologie Intégrative (GABI), Université Paris-Saclay-AgroParisTech-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Biologie de la Reproduction, Environnement, Epigénétique & Développement (BREED), École nationale vétérinaire d'Alfort (ENVA)-Université de Versailles Saint-Quentin-en-Yvelines (UVSQ)-Université Paris-Saclay-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Service de pneumologie [Hôpital Foch], Université de Paris - UFR Médecine Paris Centre [Santé] (UP Médecine Paris Centre), and Université de Paris (UP)

Subjects
Swine, medicine.medical_treatment, [SDV]Life Sciences [q-bio], Cell, Pharmacology, 03 medical and health sciences, 0302 clinical medicine, Renal Dialysis, Animals, Humans, Medicine, Lung, Dialysis, 030304 developmental biology, 0303 health sciences, Transplantation, business.industry, Cell growth, Pig model, Organ Preservation, Metabolism, 3. Good health, Perfusion, medicine.anatomical_structure, 030220 oncology & carcinogenesis, business, Ex vivo, Lung Transplantation
Abstract

International audience; Background: Normothermic ex vivo lung perfusion (EVLP) increases the pool of donor lungs by requalifying marginal lungs refused for transplantation through the recovery of macroscopic and functional properties. However the cell response and metabolism occurring during EVLP generate a nonphysiological accumulation of electrolytes, metabolites, cytokines and other cellular byproducts which may have deleterious effects both at the organ and cell levels, with impact on transplantation outcomes.Methods: We analyzed the physiological, metabolic and genome-wide response of lungs undergoing a 6-hour EVLP procedure in a pig model in 4 experimental conditions: without perfusate modification, with partial replacement of fluid, and with adult or pediatric dialysis filters.Results: Adult and pediatric dialysis stabilized the electrolytic and metabolic profiles while maintaining acid-base and gas exchanges. Pediatric dialysis increased the level of IL-10 and IL-6 in the perfusate. Despite leading to modification of the perfusate composition, the 4 EVLP conditions did not affect the gene expression profiles which were associated in all cases with increased cell survival, cell proliferation, inflammatory response and cell movement, and with inhibition of bleeding.Conclusions: Management of EVLP perfusate by periodic replacement and continuous dialysis has no significant effect on the lung function nor on the gene expression profiles ex vivo. These results suggest that the accumulation of dialysable cell products does not significantly alter the lung cell response during EVLP, a finding that may have impact on EVLP management in the clinic.

Published
2021

32. A comprehensive overview of bull sperm-borne small non-coding RNAs and their diversity across breeds

Author

Chrystelle Le Danvic, Eli Sellem, Mekki Boussaha, Hélène Kiefer, Laurent Schibler, Hélène Jammes, Luc Jouneau, Sébastien Fritz, Sylvain Marthey, Jean-Philippe Perrier, Aurelie Bonnet, Andrea Rau, Biologie de la Reproduction, Environnement, Epigénétique & Développement (BREED), École nationale vétérinaire d'Alfort (ENVA)-Université de Versailles Saint-Quentin-en-Yvelines (UVSQ)-Université Paris-Saclay-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Allice, Génétique Animale et Biologie Intégrative (GABI), Université Paris-Saclay-AgroParisTech-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), ANR-14-LAB3-0008,SeQuaMol,Qualité de la semence et fertilité chez les bovins: développement de nouveaux outils moléculaires de diagnostic(2014), and Université Paris-Saclay-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-École nationale vétérinaire d'Alfort (ENVA)-Université de Versailles Saint-Quentin-en-Yvelines (UVSQ)

Subjects
Male, endocrine system, lcsh:QH426-470, Piwi-interacting RNA, piRNA, Biology, Bull, Deep sequencing, [SDV.GEN.GPL]Life Sciences [q-bio]/Genetics/Plants genetics, 03 medical and health sciences, 0302 clinical medicine, IsomiR, [STAT.ML]Statistics [stat]/Machine Learning [stat.ML], [SDV.BBM.GTP]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Genomics [q-bio.GN], Gene expression, isomiR, Genetics, Animals, Molecular Biology, tRNA, 030304 developmental biology, miRNA, 0303 health sciences, [SDV.GEN]Life Sciences [q-bio]/Genetics, [STAT.AP]Statistics [stat]/Applications [stat.AP], urogenital system, Research, [SDV.BA]Life Sciences [q-bio]/Animal biology, Genetic Variation, RNA, [SDV.BBM.MN]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Molecular Networks [q-bio.MN], Ribosomal RNA, Spermatozoa, Sperm, [SDV.BIBS]Life Sciences [q-bio]/Quantitative Methods [q-bio.QM], [STAT]Statistics [stat], lcsh:Genetics, [SDV.GEN.GA]Life Sciences [q-bio]/Genetics/Animal genetics, [SDV.GEN.GH]Life Sciences [q-bio]/Genetics/Human genetics, 030220 oncology & carcinogenesis, Transfer RNA, sncRNA, RNA, Small Untranslated, Cattle, Transcriptome, [SDV.AEN]Life Sciences [q-bio]/Food and Nutrition, [STAT.ME]Statistics [stat]/Methodology [stat.ME]
Abstract

Background Mature sperm carry thousands of RNAs, including mRNAs, lncRNAs, tRNAs, rRNAs and sncRNAs, though their functional significance is still a matter of debate. Growing evidence suggests that sperm RNAs, especially sncRNAs, are selectively retained during spermiogenesis or specifically transferred during epididymis maturation, and are thus delivered to the oocyte at fertilization, providing resources for embryo development. However , a deep characterization of the sncRNA content of bull sperm and its expression profile across breeds is currently lacking. To fill this gap, we optimized a guanidinium–Trizol total RNA extraction protocol to prepare high-quality RNA from frozen bull sperm collected from 40 representative bulls from six breeds. Deep sequencing was performed (40 M single 50-bp reads per sample) to establish a comprehensive repertoire of cattle sperm sncRNA. Results Our study showed that it comprises mostly piRNAs (26%), rRNA fragments (25%), miRNAs (20%) and tRNA fragments (tsRNA, 14%). We identified 5p-halves as the predominant tsRNA subgroup in bull sperm, originating mostly from Gly and Glu isoacceptors. Our study also increased by ~ 50% the sperm repertoire of known miRNAs and identified 2022 predicted miRNAs. About 20% of sperm miRNAs were located within genomic clusters, expanding the list of known polycistronic pri-miRNA clusters and defining several networks of co-expressed miRNAs. Strikingly, our study highlighted the great diversity of isomiRs, resulting mainly from deletions and non-templated additions (A and U) at the 3p end. Substitutions within miRNA sequence accounted for 40% of isomiRs, with G>A, U>C and C>U substitutions being the most frequent variations. In addition, many sncRNAs were found to be differentially expressed across breeds. Conclusions Our study provides a comprehensive overview of cattle sperm sncRNA, and these findings will pave the way for future work on the role of sncRNAs in embryo development and their relevance as biomarkers of semen fertility.

Published
2020

33. Transient Changes of Metabolism at The Pronuclear Stage in Mice Influences Skeletal Muscle Phenotype in Adulthood

Author

Amélie Bonnet-Garnier, Barbara Vernus, Christelle Bertrand-Gaday, Laurence Pessemesse, Anne Bonnieu, Emilie Blanchet, François Casas, Martine Letheule, Dynamique Musculaire et Métabolisme (DMEM), Université de Montpellier (UM)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Biologie de la Reproduction, Environnement, Epigénétique & Développement (BREED), Université Paris-Saclay-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-École nationale vétérinaire d'Alfort (ENVA)-Université de Versailles Saint-Quentin-en-Yvelines (UVSQ), École nationale vétérinaire d'Alfort (ENVA), Fondation pour la Recherche Médicale, and École nationale vétérinaire d'Alfort (ENVA)-Université de Versailles Saint-Quentin-en-Yvelines (UVSQ)-Université Paris-Saclay-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)

Subjects
Male, 0301 basic medicine, [SDV]Life Sciences [q-bio], Myostatin, Mitochondrion, Muscle hypertrophy, lcsh:Chemistry, Mice, 0302 clinical medicine, Pyruvic Acid, skeletal Muscle, metabolic reprogramming, Glycolysis, lcsh:QH301-705.5, Spectroscopy, 030219 obstetrics & reproductive medicine, biology, Acetylation, Embryo, General Medicine, Phenotype, Computer Science Applications, Cell biology, mitochondria, medicine.anatomical_structure, Carbohydrate Metabolism, Cytokines, Female, Oxidation-Reduction, epigenetic, Muscle Contraction, Genotype, Embryonic Development, embryo, Article, Catalysis, Inorganic Chemistry, 03 medical and health sciences, medicine, Animals, Epigenetics, Physical and Theoretical Chemistry, Muscle, Skeletal, Molecular Biology, Organic Chemistry, Skeletal muscle, Hypertrophy, Embryo, Mammalian, Disease Models, Animal, 030104 developmental biology, lcsh:Biology (General), lcsh:QD1-999, biology.protein
Abstract

Skeletal muscle has a remarkable plasticity, and its phenotype is strongly influenced by hormones, transcription factors, and physical activity. However, whether skeletal phenotype can be oriented or not during early embryonic stages has never been investigated. Here, we report that pyruvate as the only source of carbohydrate in the culture medium of mouse one cell stage embryo influenced the establishment of the muscular phenotype in adulthood. We found that pyruvate alone induced changes in the contractile phenotype of the skeletal muscle in a sexually dependent manner. For male mice, a switch to a more glycolytic phenotype was recorded, whereas, in females, the pyruvate induced a switch to a more oxidative phenotype. In addition, the influence of pyruvate on the contractile phenotypes was confirmed in two mouse models of muscle hypertrophy: the well-known myostatin deficient mouse (Mstn&minus, /&minus, ) and a mouse carrying a specific deletion of p43, a mitochondrial triiodothyronine receptor. Finally, to understand the link between these adult phenotypes and the early embryonic period, we assessed the levels of two histone H3 post-translational modifications in presence of pyruvate alone just after the wave of chromatin reprogramming specific of the first cell cycle. We showed that H3K4 acetylation level was decreased in Mstn&minus, 2-cell embryos, whereas no difference was found for H3K27 trimethylation level, whatever the genotype. These findings demonstrate for the first time that changes in the access of energy substrate during the very first embryonic stage can induce a precocious orientation of skeletal muscle phenotype in adulthood.

Published
2020

34. Prenatal cannabinoid exposure alters the ovarian reserve in adult offspring of rats

Author

Hubert Lepidi, Anne-Laure Pelissier-Alicot, Beatrice Mandon-Pepin, Blandine Courbiere, Magalie Barbier, Elodie Poumerol, Pierre Castel, Olivier J. Manzoni, Virginie Tassistro, Service de gynécologie-obstétrique [Hôpital Nord - APHM], Aix Marseille Université (AMU)-Assistance Publique - Hôpitaux de Marseille (APHM)- Hôpital Nord [CHU - APHM], Institut méditerranéen de biodiversité et d'écologie marine et continentale (IMBE), Avignon Université (AU)-Aix Marseille Université (AMU)-Institut de recherche pour le développement [IRD] : UMR237-Centre National de la Recherche Scientifique (CNRS), Biologie de la Reproduction, Environnement, Epigénétique & Développement (BREED), Université Paris-Saclay-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-École nationale vétérinaire d'Alfort (ENVA)-Université de Versailles Saint-Quentin-en-Yvelines (UVSQ), Laboratoire d'anatomie pathologique - [Hôpital Nord - APHM], Hôpital Nord [CHU - APHM]-Aix Marseille Université (AMU)-Assistance Publique - Hôpitaux de Marseille (APHM), Microbes évolution phylogénie et infections (MEPHI), Institut de Recherche pour le Développement (IRD)-Aix Marseille Université (AMU)-Centre National de la Recherche Scientifique (CNRS), Institut de Neurobiologie de la Méditerranée [Aix-Marseille Université] (INMED - INSERM U1249), Institut National de la Santé et de la Recherche Médicale (INSERM)-Aix Marseille Université (AMU), A*Midex Foundation of Aix-Marseille University, École nationale vétérinaire - Alfort (ENVA)-Université de Versailles Saint-Quentin-en-Yvelines (UVSQ)-Université Paris-Saclay-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), and Aix Marseille Université (AMU)-Institut National de la Santé et de la Recherche Médicale (INSERM)

Subjects
0301 basic medicine, Agonist, Endocannabinoid system, Drug Inverse Agonism, medicine.drug_class, Offspring, Health, Toxicology and Mutagenesis, medicine.medical_treatment, Morpholines, Gestational Age, 010501 environmental sciences, Biology, Naphthalenes, Toxicology, 01 natural sciences, Andrology, 03 medical and health sciences, Receptor, Cannabinoid, CB1, Pregnancy, Prenatal exposure, SR141716, medicine, Delta 9-tetrahydrocannabinol, Inverse agonist, Animals, Dronabinol, Young adult, Rats, Wistar, Ovarian reserve, Ovarian Reserve, Cannabinoid Receptor Antagonists, 0105 earth and related environmental sciences, Cannabinoid Receptor Agonists, [SDV.BA]Life Sciences [q-bio]/Animal biology, Ovary, General Medicine, Benzoxazines, 030104 developmental biology, Gene Expression Regulation, Prenatal Exposure Delayed Effects, Gestation, Female, Cannabinoid, Rimonabant, WIN55212, Endocannabinoids
Abstract

International audience; In animals, research in the past two decades has demonstrated the strong involvement of the endocannabinoid system (ECS) in numerous steps of the reproductive process, including ovarian physiology. Reproductive lifespan is closely related to the number of nongrowing ovarian follicles, called ovarian reserve (OR), which is definitively established during foetal life. Thus, OR damage may lead to poor reproductive outcomes and a shortened reproductive lifespan. We investigated whether prenatal ECS modulation had an effect on the OR at different ages in the rat offspring. Four groups of gestating female rats (F0) were exposed to the CB1-/CB2-receptor agonist WIN55212 (0.5 mg/kg), the CB1R inverse agonist SR141716 (3 mg/kg) or Δ9THC (5 mg/kg) and were compared to negative control groups. OR was histologically assessed at different postnatal timepoints (F1 individuals): postnatal day (PND) 6, PND40 and PND90. At PND6, prenatal exposure had no effect on OR. In the young adult group (PND90) exposed during gestation to WIN55212, we observed a CB1R-mediated delayed OR decrease, which was reversed by prenatal CB1R blockade by SR141716. Conversely, after prenatal SR141716 exposure, we observed higher OR counts at PND90. RT-PCR experiments also showed that prenatal ECS modulation perturbed the mRNA levels of ECS enzymes and OR regulation genes. Our findings support the role of the ECS in OR regulation during the foetal life of rats and highlight the need for further studies to elucidate its precise role in OR physiology.

Published
2020

35. Ephrin-B2 paces neuronal production in the developing neocortex

Author

Anthony Kischel, Christophe Audouard, Alice Davy, Mohamad-Ali Fawal, Centre de Biologie Intégrative (CBI), Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique (CNRS), Centre de biologie du développement (CBD), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique (CNRS)-Centre de Biologie Intégrative (CBI), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS), Biologie de la Reproduction, Environnement, Epigénétique & Développement (BREED), Université Paris-Saclay-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-École nationale vétérinaire d'Alfort (ENVA)-Université de Versailles Saint-Quentin-en-Yvelines (UVSQ), Universite Paul Sabatier Centre National de la Recherche Scientifique (CNRS), and ANR: French National Research Agency ,15CE13-0010-01

Subjects
Male, animal structures, Mouse, Neurogenesis, [SDV]Life Sciences [q-bio], Blotting, Western, Fluorescent Antibody Technique, Neocortex, Ephrin-B2, Biology, 03 medical and health sciences, Mice, 0302 clinical medicine, medicine, Eph:ephrin signaling, Ephrin, Animals, lcsh:QH301-705.5, [SDV.BDD]Life Sciences [q-bio]/Development Biology, 030304 developmental biology, ephrin signaling, Neurons, 0303 health sciences, Cell Cycle, Erythropoietin-producing hepatocellular (Eph) receptor, Cell Differentiation, Embryonic stem cell, Juxtacrine signalling, Neural progenitors, biological factors, Eph, medicine.anatomical_structure, [SDV.BDD.EO]Life Sciences [q-bio]/Development Biology/Embryology and Organogenesis, lcsh:Biology (General), nervous system, Cerebral cortex, embryonic structures, Female, Neuron, sense organs, Neuroscience, 030217 neurology & neurosurgery, Developmental Biology, Research Article, Signal Transduction
Abstract

Background During mammalian cerebral cortex development, different types of projection neurons are produced in a precise temporal order and in stereotypical numbers. The mechanisms regulating timely generation of neocortex projection neurons and ensuring production in sufficient numbers of each neuronal identity are only partially understood. Results Here, we show that ephrin-B2, a member of the Eph:ephrin cell-to-cell communication pathway, sets the neurogenic tempo in the neocortex. Indeed, conditional mutant embryos for ephrin-B2 exhibit a transient delay in neurogenesis and acute stimulation of Eph signaling by in utero injection of synthetic ephrin-B2 led to a transient increase in neuronal production. Using genetic approaches we show that ephrin-B2 acts on neural progenitors to control their differentiation in a juxtacrine manner. Unexpectedly, we observed that perinatal neuron numbers recovered following both loss and gain of ephrin-B2, highlighting the ability of neural progenitors to adapt their behavior to the state of the system in order to produce stereotypical numbers of neurons. Conclusions Altogether, our data uncover a role for ephrin-B2 in embryonic neurogenesis and emphasize the plasticity of neuronal production in the neocortex.

Published
2020

36. Differential Effects of Post-Weaning Diet and Maternal Obesity on Mouse Liver and Brain Metabolomes

Author

Christine Baly, Hélène Jammes, Delphine Rousseau-Ralliard, Luc Jouneau, Alexandre Seyer, Etienne A. Thévenot, Sofiane Safi-Stibler, Anne Gabory, Mélanie Jouin, Biologie de la Reproduction, Environnement, Epigénétique & Développement (BREED), École nationale vétérinaire - Alfort (ENVA)-Université de Versailles Saint-Quentin-en-Yvelines (UVSQ)-Université Paris-Saclay-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), École nationale vétérinaire - Alfort (ENVA), Collège Doctoral, Sorbonne Université (SU), Laboratoire Sciences des Données et de la Décision (LS2D), Département Métrologie Instrumentation & Information (DM2I), Laboratoire d'Intégration des Systèmes et des Technologies (LIST (CEA)), Direction de Recherche Technologique (CEA) (DRT (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Direction de Recherche Technologique (CEA) (DRT (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Laboratoire d'Intégration des Systèmes et des Technologies (LIST (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay, Laboratoire d'Etude du Métabolisme des Médicaments (LEMM), Service de Pharmacologie et Immunoanalyse (SPI), Médicaments et Technologies pour la Santé (MTS), Université Paris-Saclay-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Université Paris-Saclay-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Médicaments et Technologies pour la Santé (MTS), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), MedDay Pharmaceuticals, FCA 13T1 - Ministère de l'Enseignement supérieur, de la Recherche et de l'Innovation, MESRI, Fondation Coeur et Artères FCA 13T1- French National Research Institute for Agriculture, Food and Environment (INRAE, Institut National de Recherche pour l'Agriculture, l'Alimentation et l'Environnement), through the AlimH 'omics' program 2016 - Ministry of Research, France, École nationale vétérinaire d'Alfort (ENVA)-Université de Versailles Saint-Quentin-en-Yvelines (UVSQ)-Université Paris-Saclay-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), École nationale vétérinaire d'Alfort (ENVA), Collège doctoral [Sorbonne universités], Laboratoire d'Intégration des Systèmes et des Technologies (LIST), and Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)

Subjects
0301 basic medicine, Male, obesity, Physiology, Energy homeostasis, Obesity, Maternal, 0302 clinical medicine, Weight loss, Pregnancy, Homeostasis, Anserine, Maternal-Fetal Exchange, Nutrition and Dietetics, 3-Hydroxybutyric Acid, DOHaD, Brain, metabolomics, high-fat diet, Hypothalamus, Statistical analysis, Prenatal Exposure Delayed Effects, Metabolome, Female, medicine.symptom, lcsh:Nutrition. Foods and food supply, Offspring, lcsh:TX341-641, Weaning, Biology, Deoxyglucose, liver, Article, 03 medical and health sciences, Metabolomics, [SDV.BBM.GTP]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Genomics [q-bio.GN], medicine, Animals, Lysine, Maternal Nutritional Physiological Phenomena, medicine.disease, Obesity, Diet, Mice, Inbred C57BL, Metabolic pathway, 030104 developmental biology, weight loss, Energy Metabolism, [SDV.AEN]Life Sciences [q-bio]/Food and Nutrition, 030217 neurology & neurosurgery, Food Science
Abstract

Nutritional changes during developmental windows are of particular concern in offspring metabolic disease. Questions are emerging concerning the role of maternal weight changes before conception, particularly for weight loss, in the development of diet-related disorders. Understanding the physiological pathways affected by the maternal trajectories in the offspring is therefore essential, but a broad overview is still lacking. We recently reported both metabolic and behavioral negative outcomes in offspring born to obese or weight-loss mothers and fed a control of high-fat diet, suggesting long-term modeling of metabolic pathways needing to be further characterized. Using non-targeted LC&ndash, HRMS, we investigated the impact of maternal and post-weaning metabolic status on the adult male offspring&rsquo, s metabolome in three tissues involved in energy homeostasis: liver, hypothalamus and olfactory bulb. We showed that post-weaning diet interfered with the abundance of several metabolites, including 1,5-anhydroglucitol, saccharopine and &beta, hydroxybutyrate, differential in the three tissues. Moreover, maternal diet had a unique impact on the abundance of two metabolites in the liver. Particularly, anserine abundance, lowered by maternal obesity, was normalized by a preconceptional weight loss, whatever the post-weaning diet. This study is the first to identify a programming long-term effect of maternal preconception obesity on the offspring metabolome.

Published
2020

37. No-contact microchip measurements of body temperature and behavioural changes prior to foaling

Author

Florence Jaffrezic, Tristan Jousset, Juliette Auclair-Ronzaud, Pascale Chavatte-Palmer, Cédric Dubois, Laurence Wimel, IFCE, Plateau Technique de la Station Expérimentale, Biologie de la Reproduction, Environnement, Epigénétique & Développement (BREED), and École nationale vétérinaire d'Alfort (ENVA)-Université de Versailles Saint-Quentin-en-Yvelines (UVSQ)-Université Paris-Saclay-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)

Subjects
Flehmen response, TEMPERATURE DECREASE, Temperature monitoring, Rump, animal diseases, Microchip, Body Temperature, 03 medical and health sciences, 0302 clinical medicine, Animal science, Food Animals, Pregnancy, biology.animal, Medicine, Animals, Behaviour, Horses, Video monitoring, Small Animals, 030219 obstetrics & reproductive medicine, [SDV.BA.MVSA]Life Sciences [q-bio]/Animal biology/Veterinary medicine and animal Health, Sheep, biology, business.industry, Equine, 0402 animal and dairy science, Parturition, Temperature, Horse, 04 agricultural and veterinary sciences, 040201 dairy & animal science, Accelerometer, Foal, Animal Science and Zoology, Cattle, Female, business, Gestational length
Abstract

International audience; estational length is highly variable in horses ranging from 320 to 360 days. Thus, determining parturition time is an important challenge for the horse industry. Body temperature can be used in cows and ewes as an indicator of parturition. Thus, the aim of this study is to determine if temperature can also be used as indicator of foaling. Thirty-nine mares were monitored over two foaling seasons (2018 and 2019). They were housed in 16 m2 stalls with access to pasture in group three times a week from 10:00 to 16:00. Night watch as well as video monitoring was ensured during foaling periods. Body temperature was monitored using an identification and temperature sensor microchip implanted in the neckline. Measurement were taken manually every 2 h from 5 days before to 6 h after parturition by moving a microchip reader close to the mares' neck. Mares were equipped with a tail accelerometer recording tail movements and lateral recumbency 24 h before parturition. In addition, behaviour was monitored by video analysis in the hour preceding expulsion of the foal in 8 individuals in 2019. Relationships between behavioural and temperature data were explored throughout principal component analysis (PCA). All foals were born healthy and no human intervention was required during foaling. Mean daily body temperature decreased significantly by 0.3 °C (95%; range: 0.42 to -0.19 °C) between the day of parturition and the mean temperature of the 5 preceding days. A significant temperature decrease was also detected 12 h before and at the onset of parturition. With a 0.5 °C threshold, foaling could be detected 12 h in advance with 96.6% sensitivity and 95.0% specificity, respectively. Tail movements were more frequent and shorter with impending parturition. Body temperature was positively correlated with increased frequency and duration of specific behaviours (flehmen, looking at their flank and rump scratching against the stall wall). In conclusion, as in other species, body temperature was related to signs usually associated with impeding parturition, with a significant temperature drop observed from 12 h before and at the time of foaling. Providing automated measurements become available, temperature monitoring could become an additional tool to predict parturition in mares.

Published
2020

38. Dynamic CpG methylation delineates subregions within super-enhancers selectively decommissioned at the exit from naive pluripotency

Author

Edward Curry, Véronique Azuara, King Hang T. Mau, Roshni A. de Souza, Vincent Brochard, Alice Jouneau, Rute A. Tomaz, Luc Jouneau, Wout Megchelenbrink, Emma Bell, Hendrik Marks, Yaser Atlasi, Hendrik G. Stunnenberg, Bell, Emma [0000-0003-4103-3741], Tomaz, Rute A [0000-0002-9377-1431], Marks, Hendrik [0000-0002-4198-3731], Stunnenberg, Hendrik G [0000-0002-0066-1576], Azuara, Véronique [0000-0003-4608-3713], Apollo - University of Cambridge Repository, Biologie du développement et reproduction (BDR), École nationale vétérinaire d'Alfort (ENVA)-Institut National de la Recherche Agronomique (INRA)-Centre National de la Recherche Scientifique (CNRS), Imperial College London, Radboud Institute for Molecular Life Sciences (RIMLS), Unité de recherche Virologie et Immunologie Moléculaires (VIM), Institut National de la Recherche Agronomique (INRA), Medical Research Council (MRC), Genesis Research Trust, The Portuguese Science & Technology foundation (FCT), Hammersmith Hospital, Faculty of Medicine and Health, Institute of Reproductive Sciences (IRS), Imperial College, London, Princess Máxima Center for Pediatric Oncology, Centre for Molecular Oncology, Department of Precision Medicine, Università della Campania Luigi Vanvitelli, Université Paris Saclay (COMUE), Institute of Reproductive and Developmental Biology, Faculty of Medicine, Department of Molecular Biology, Princeton University, Faculty of Health, Medicine and Life Sciences, Biologie de la Reproduction, Environnement, Epigénétique & Développement (BREED), École nationale vétérinaire d'Alfort (ENVA)-Université de Versailles Saint-Quentin-en-Yvelines (UVSQ)-Université Paris-Saclay-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Biologie du Développement et Reproduction (BDR), École nationale vétérinaire d'Alfort (ENVA)-Institut National de la Recherche Agronomique (INRA), Unité de recherche Virologie et Immunologie Moléculaires (VIM (UR 0892)), French Society for Stem Cell Research. Corbeil-Essonne, FRA., Bell, E., Curry, E. W., Megchelenbrink, W., Jouneau, L., Brochard, V., Tomaz, R. A., Mau, K. H. T., Atlasi, Y., de Souza, R. A., Marks, H., Stunnenberg, H. G., Jouneau, A., Azuara, V., Jouneau, Alice, Azuara, Véronique, and ProdInra, Archive Ouverte

Subjects
0301 basic medicine, CHROMATIN, Transcription, Genetic, [SDV]Life Sciences [q-bio], General Physics and Astronomy, Stem cells, Mice, 0302 clinical medicine, BINDING, MEDIATOR, lcsh:Science, méthylation, Induced pluripotent stem cell, [SDV.BDD]Life Sciences [q-bio]/Development Biology, Regulation of gene expression, Multidisciplinary, Mediator Complex, Gene Expression Regulation, Developmental, Mouse Embryonic Stem Cells, Cell biology, Chromatin, Multidisciplinary Sciences, TRANSCRIPTION FACTORS, Enhancer Elements, Genetic, CpG site, Receptors, Estrogen, GROUND-STATE, DNA methylation, embryonic structures, Science & Technology - Other Topics, RNA Polymerase II, TRANSITION, Germ Layers, Protein Binding, musculoskeletal diseases, Pluripotent Stem Cells, Embryonic stem cells, Science, education, chemical and pharmacologic phenomena, Biology, Article, General Biochemistry, Genetics and Molecular Biology, 03 medical and health sciences, [SDV.BDD] Life Sciences [q-bio]/Development Biology, Developmental biology, parasitic diseases, pluripotence, Animals, enhancer, facteur de transcription, natural sciences, Enhancer, Molecular Biology, Science & Technology, General Chemistry, DNA Methylation, pluripotency, Embryonic stem cell, GENE, 030104 developmental biology, nervous system, Epiblast, IDENTITY, lcsh:Q, CpG Islands, EMBRYONIC STEM-CELLS, 030217 neurology & neurosurgery
Abstract

Clusters of enhancers, referred as to super-enhancers (SEs), control the expression of cell identity genes. The organisation of these clusters, and how they are remodelled upon developmental transitions remain poorly understood. Here, we report the existence of two types of enhancer units within SEs typified by distinctive CpG methylation dynamics in embryonic stem cells (ESCs). We find that these units are either prone for decommissioning or remain constitutively active in epiblast stem cells (EpiSCs), as further established in the peri-implantation epiblast in vivo. Mechanistically, we show a pivotal role for ESRRB in regulating the activity of ESC-specific enhancer units and propose that the developmentally regulated silencing of ESRRB triggers the selective inactivation of these units within SEs. Our study provides insights into the molecular events that follow the loss of ESRRB binding, and offers a mechanism by which the naive pluripotency transcriptional programme can be partially reset upon embryo implantation., Clusters of enhancers, called super-enhancers (SEs), control the expression of cell identity genes. Here, the authors identify two types of enhancer units within SEs in ESCs, characterised by distinctive CpG methylation dynamics, and find that ESRRB regulates the enhancer units decommissioned at exit from naive pluripotency.

Published
2020

39. Retinoic acid synthesis by ALDH1A proteins is dispensable for meiosis initiation in the mouse fetal ovary

Author

Morgane Le Rolle, Isabelle Stévant, Andreas Schedl, Marie-Christine Chaboissier, Norbert B. Ghyselinck, Eric Pailhoux, Fabio Da Silva, Geneviève Jolivet, Anne-Amandine Chassot, Jean-Marie Guigonis, Serge Nef, Institut de Biologie Valrose (IBV), Université Nice Sophia Antipolis (... - 2019) (UNS), COMUE Université Côte d'Azur (2015-2019) (COMUE UCA)-COMUE Université Côte d'Azur (2015-2019) (COMUE UCA)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université Côte d'Azur (UCA), Biologie de la Reproduction, Environnement, Epigénétique & Développement (BREED), École nationale vétérinaire d'Alfort (ENVA)-Université de Versailles Saint-Quentin-en-Yvelines (UVSQ)-Université Paris-Saclay-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Université de Genève (UNIGE), Université Côte d'Azur - Faculté de Médecine (UCA Faculté Médecine), Université Côte d'Azur (UCA), Institut de Génétique et de Biologie Moléculaire et Cellulaire (IGBMC), Université de Strasbourg (UNISTRA)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), COMUE Université Côte d'Azur (2015-2019) (COMUE UCA)-COMUE Université Côte d'Azur (2015-2019) (COMUE UCA)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Côte d'Azur (UCA)-Centre National de la Recherche Scientifique (CNRS), Université Paris-Saclay-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-École nationale vétérinaire d'Alfort (ENVA)-Université de Versailles Saint-Quentin-en-Yvelines (UVSQ), Medical School of the University of Geneva, German Cancer Research Center - Deutsches Krebsforschungszentrum [Heidelberg] (DKFZ), French National Research Agency (ANR)ANR-13-BSV2-0017-02 ARGONADSANR-11-LABX-0028-01ANR-18-CE14-0012 SexSpecsLa Ligue Nationale Contre le Cancer (Equipe Labelisée Ligue Nationale Contre le Cancer), ANR-13-BSV2-0017,ARGONADS,Acide rétinoique et devenir des cellules germinales(2013), ANR-18-CE14-0012,Sex-Specs,Homeostasie tissulaire selon le sex et son impact sur les maladies surrénaliennes(2018), COMUE Université Côte d'Azur (2015 - 2019) (COMUE UCA)-COMUE Université Côte d'Azur (2015 - 2019) (COMUE UCA)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université Côte d'Azur (UCA), UMR E4320 (TIRO-MATOs), COMUE Université Côte d'Azur (2015 - 2019) (COMUE UCA)-COMUE Université Côte d'Azur (2015 - 2019) (COMUE UCA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Côte d'Azur (UCA), Department of Genetic Medicine and Development [Geneva], Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA), Chaboissier, Marie-Christine, Blanc 2013 - Acide rétinoique et devenir des cellules germinales - - ARGONADS2013 - ANR-13-BSV2-0017 - Blanc 2013 - VALID, APPEL À PROJETS GÉNÉRIQUE 2018 - Homeostasie tissulaire selon le sex et son impact sur les maladies surrénaliennes - - Sex-Specs2018 - ANR-18-CE14-0012 - AAPG2018 - VALID, Centres d'excellences - Réseau d'Innovation sur les Voies de Signalisation en Sciences de la Vie - - SIGNALIFE2011 - ANR-11-LABX-0028 - LABX - VALID, Université Nice Sophia Antipolis (1965 - 2019) (UNS), École nationale vétérinaire - Alfort (ENVA)-Université de Versailles Saint-Quentin-en-Yvelines (UVSQ)-Université Paris-Saclay-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), COMUE Université Côte d'Azur (2015-2019) (COMUE UCA)-COMUE Université Côte d'Azur (2015-2019) (COMUE UCA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Côte d'Azur (UCA), ANR-13-BSV2-0017-02 ARGONADSANR-11-LABX-0028-01ANR-18-CE14-0012 SexSpecsLa Ligue Nationale Contre le Cancer (Equipe Labelisée), ANR-11-LABX-0028,SIGNALIFE,Réseau d'Innovation sur les Voies de Signalisation en Sciences de la Vie(2011), and COMUE Université Côte d'Azur (2015 - 2019) (COMUE UCA)-COMUE Université Côte d'Azur (2015 - 2019) (COMUE UCA)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Côte d'Azur (UCA)-Centre National de la Recherche Scientifique (CNRS)

Subjects
[SDV]Life Sciences [q-bio], Retinoic acid, Endogeny, Oogenesis, chemistry.chemical_compound, 0302 clinical medicine, CYP26B1, PRIMORDIAL GERM-CELLS, BINDING, ddc:576.5, [SDV.BDD]Life Sciences [q-bio]/Development Biology, ComputingMilieux_MISCELLANEOUS, Research Articles, Mammals, 0303 health sciences, Multidisciplinary, integumentary system, SciAdv r-articles, CELL SEX DETERMINATION, LINEAGE, MEIOSIS, OOGENESIS, Cell biology, [SDV] Life Sciences [q-bio], medicine.anatomical_structure, DIFFERENTIATION, Female, Research Article, EXPRESSION, FATE, Ovary, Tretinoin, Biology, ALDH1A2, 03 medical and health sciences, Meiosis, medicine, Animals, Gene, neoplasms, 030304 developmental biology, organic chemicals, Proteins, Cell Biology, GENE, biological factors, ALDH1A1, body regions, MICE, Germ Cells, chemistry, biology.protein, 030217 neurology & neurosurgery, Developmental Biology
Abstract

Retinoic acid synthesis is not essential for meiosis in fetal ovaries, challenging the dogma about the meiosis-inducing factors., In mammals, the timing of meiosis entry is regulated by signals from the gonadal environment. All-trans retinoic acid (ATRA) signaling is considered the key pathway that promotes Stra8 (stimulated by retinoic acid 8) expression and, in turn, meiosis entry. This model, however, is debated because it is based on analyzing the effects of exogenous ATRA on ex vivo gonadal cultures, which not accurately reflects the role of endogenous ATRA. Aldh1a1 and Aldh1a2, two retinaldehyde dehydrogenases synthesizing ATRA, are expressed in the mouse ovaries when meiosis initiates. Contrary to the present view, here, we demonstrate that ATRA-responsive cells are scarce in the ovary. Using three distinct gene deletion models for Aldh1a1;Aldh1a2;Aldh1a3, we show that Stra8 expression is independent of ATRA production by ALDH1A proteins and that germ cells progress through meiosis. Together, these data demonstrate that ATRA signaling is dispensable for instructing meiosis initiation in female germ cells.

Published
2020

40. Sources of variation of DNA methylation in rainbow trout: combined effects of temperature and genetic background

Author

Alain Roulet, Lionel Goardon, Céline Ciobotaru, Jean-Michel Le Calvez, Maria Bernard, Mathieu Charles, Audrey Prézelin, Marjorie Bideau, Olivier Bouchez, Laurent Labbé, Nicolas Dechamp, Edwige Quillet, Xavier Cousin, Alexandre Fricot, Mathilde Dupont-Nivet, Marco Moroldo, Delphine Lallias, Génétique Animale et Biologie Intégrative (GABI), Université Paris-Saclay-AgroParisTech-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Système d'Information des GENomes des Animaux d'Elevage (SIGENAE), Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Pisciculture Expérimentale INRAE des Monts d'Arrée (PEIMA), Biologie de la Reproduction, Environnement, Epigénétique & Développement (BREED), Université Paris-Saclay-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-École nationale vétérinaire d'Alfort (ENVA)-Université de Versailles Saint-Quentin-en-Yvelines (UVSQ), Génome et Transcriptome - Plateforme Génomique ( GeT-PlaGe), Plateforme Génome & Transcriptome (GET), Génopole Toulouse Midi-Pyrénées [Auzeville] (GENOTOUL), Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Ecole Nationale Vétérinaire de Toulouse (ENVT), Institut National Polytechnique (Toulouse) (Toulouse INP), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Institut National Polytechnique (Toulouse) (Toulouse INP), Université Fédérale Toulouse Midi-Pyrénées-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), Université Fédérale Toulouse Midi-Pyrénées-Institut National de la Santé et de la Recherche Médicale (INSERM)-Génopole Toulouse Midi-Pyrénées [Auzeville] (GENOTOUL), Université Fédérale Toulouse Midi-Pyrénées-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), H2020 European Research Council652831, European Project: 652831,H2020,H2020-INFRAIA-2014-2015,AQUAEXCEL2020(2015), AgroParisTech-Université Paris-Saclay-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), École nationale vétérinaire - Alfort (ENVA)-Université de Versailles Saint-Quentin-en-Yvelines (UVSQ)-Université Paris-Saclay-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), MARine Biodiversity Exploitation and Conservation (UMR MARBEC), Institut de Recherche pour le Développement (IRD)-Institut Français de Recherche pour l'Exploitation de la Mer (IFREMER)-Université de Montpellier (UM)-Centre National de la Recherche Scientifique (CNRS), Université Toulouse III - Paul Sabatier (UT3), Université de Toulouse (UT)-Université de Toulouse (UT)-Ecole Nationale Vétérinaire de Toulouse (ENVT), Université de Toulouse (UT)-Université de Toulouse (UT)-Institut National Polytechnique (Toulouse) (Toulouse INP), Université de Toulouse (UT)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Université Toulouse III - Paul Sabatier (UT3), Université de Toulouse (UT)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Génopole Toulouse Midi-Pyrénées [Auzeville] (GENOTOUL), Université de Toulouse (UT)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), École nationale vétérinaire d'Alfort (ENVA)-Université de Versailles Saint-Quentin-en-Yvelines (UVSQ)-Université Paris-Saclay-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Ecole Nationale Vétérinaire de Toulouse (ENVT), Université Fédérale Toulouse Midi-Pyrénées-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), Université Fédérale Toulouse Midi-Pyrénées-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Génopole Toulouse Midi-Pyrénées [Auzeville] (GENOTOUL), Université Fédérale Toulouse Midi-Pyrénées-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), and Université Fédérale Toulouse Midi-Pyrénées-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Université Toulouse III - Paul Sabatier (UT3)

Subjects
0301 basic medicine, Cancer Research, [SDE.MCG]Environmental Sciences/Global Changes, Embryonic Development, Biology, DNA methyltransferase, 03 medical and health sciences, chemistry.chemical_compound, 0302 clinical medicine, Animals, isogenic lines, 14. Life underwater, Epigenetics, Genetic variability, dnmt3, Molecular Biology, Gene, Genetics, DNA methylation, EpiRADseq, Temperature, [SDV.BBM.BM]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Molecular biology, Methylation, rainbow trout, [SDV.GEN.GA]Life Sciences [q-bio]/Genetics/Animal genetics, 030104 developmental biology, chemistry, 030220 oncology & carcinogenesis, Oncorhynchus mykiss, Rainbow trout, Genetic Background, DNA, Research Paper
Abstract

International audience; Phenotypic plasticity is a key component of the ability of organisms to respond to changing environmental conditions. In this study, we aimed to study the establishment of DNA methylation marks in response to an environmental stress in rainbow trout and to assess whether these marks depend on the genetic background. The environmental stress chosen here was temperature, a known induction factor of epigenetic marks in fish. To disentangle the role of epigenetic mechanisms such as DNA methylation in generating phenotypic variations, nine rainbow trout isogenic lines with no genetic variability within a line were used. For each line, half of the eggs were incubated at standard temperature (11°C) and the other half at high temperature (16°C), from eyed-stage to hatching. In order to gain a first insight into the establishment of DNA methylation marks in response to an early temperature regime (control 11°C vs. heated 16°C), we have studied the expression of 8 dnmt3 (DNA methyltransferase) genes, potentially involved in de novo methylation, and analysed global DNA methylation in the different rainbow trout isogenic lines using LUMA (LUminometric Methylation Assay). Finally, finer investigation of genome-wide methylation patterns was performed using EpiRADseq, a reduced-representation library approach based on the ddRADseq (Double Digest Restriction Associated DNA) protocol, for six rainbow trout isogenic lines. We have demonstrated that thermal history during embryonic development alters patterns of DNA methylation, but to a greater or lesser extent depending on the genetic background.

Published
2020
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41. Identification of 56 Proteins Involved in Embryo–Maternal Interactions in the Bovine Oviduct

Author

Emmanuelle Com, Charles Banliat, Ana-Paula Teixeira-Gomes, Marie Saint-Dizier, Régis Lavigne, Charles Pineau, Pascal Mermillod, Valérie Labas, Benoit Guyonnet, Guillaume Tsikis, Physiologie de la reproduction et des comportements [Nouzilly] (PRC), Institut Français du Cheval et de l'Equitation [Saumur]-Université de Tours-Centre National de la Recherche Scientifique (CNRS)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Chirurgie et Imagerie pour la Recherche et l'Enseignement [Nouzilly] (Plate-forme CIRE), Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Infectiologie et Santé Publique (UMR ISP), Université de Tours-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Institut de recherche en santé, environnement et travail (Irset), Université d'Angers (UA)-Université de Rennes 1 (UR1), Université de Rennes (UNIV-RENNES)-Université de Rennes (UNIV-RENNES)-École des Hautes Études en Santé Publique [EHESP] (EHESP)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Structure Fédérative de Recherche en Biologie et Santé de Rennes ( Biosit : Biologie - Santé - Innovation Technologique ), Proteomics Core Facility (Protim), Université de Rennes 1 (UR1), Université de Rennes (UNIV-RENNES)-Université de Rennes (UNIV-RENNES)-Plateforme Génomique Santé Biogenouest®, Biologie de la Reproduction, Environnement, Epigénétique & Développement (BREED), Université Paris-Saclay-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-École nationale vétérinaire d'Alfort (ENVA)-Université de Versailles Saint-Quentin-en-Yvelines (UVSQ), No grant number, Allan Wilson Centre, 2017/0684, Association Nationale de la Recherche et de la Technologie, Centre National de la Recherche Scientifique (CNRS)-Université de Tours-Institut Français du Cheval et de l'Equitation [Saumur]-Institut National de la Recherche Agronomique (INRA), Biologie du Développement et Reproduction (BDR), Institut National de la Recherche Agronomique (INRA), Institut Français du Cheval et de l'Equitation [Saumur] (IFCE)-Université de Tours (UT)-Centre National de la Recherche Scientifique (CNRS)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Université de Tours (UT)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Université d'Angers (UA)-Université de Rennes (UR)-École des Hautes Études en Santé Publique [EHESP] (EHESP)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Structure Fédérative de Recherche en Biologie et Santé de Rennes ( Biosit : Biologie - Santé - Innovation Technologique ), Université de Rennes (UR)-Plateforme Génomique Santé Biogenouest®, École nationale vétérinaire - Alfort (ENVA)-Université de Versailles Saint-Quentin-en-Yvelines (UVSQ)-Université Paris-Saclay-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Institut Français du Cheval et de l'Equitation [Saumur]-Université de Tours (UT)-Centre National de la Recherche Scientifique (CNRS)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Structure Fédérative de Recherche en Biologie et Santé de Rennes ( Biosit : Biologie - Santé - Innovation Technologique )-Institut National de la Santé et de la Recherche Médicale (INSERM)-École des Hautes Études en Santé Publique [EHESP] (EHESP)-Université de Rennes 1 (UR1), Université de Rennes (UNIV-RENNES)-Université de Rennes (UNIV-RENNES)-Université d'Angers (UA), École nationale vétérinaire d'Alfort (ENVA)-Université de Versailles Saint-Quentin-en-Yvelines (UVSQ)-Université Paris-Saclay-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), and Jonchère, Laurent

Subjects
0301 basic medicine, Blastomeres, Proteome, oviduct, [SDV]Life Sciences [q-bio], Oviducts, morula, Proteomics, lcsh:Chemistry, 0302 clinical medicine, tubal fluid, OVGP1, Zona pellucida, lcsh:QH301-705.5, Spectroscopy, ComputingMilieux_MISCELLANEOUS, fallopian tube, 030219 obstetrics & reproductive medicine, bovine, Serine Endopeptidases, Embryo, General Medicine, Computer Science Applications, Cell biology, [SDV] Life Sciences [q-bio], medicine.anatomical_structure, embryonic structures, 4–6 cell, Oviduct, Female, Vitelline Membrane, endocrine system, animal structures, Annexins, embryo, Perivitelline space, Biology, Catalysis, Article, Protein–protein interaction, Inorganic Chemistry, 03 medical and health sciences, proteomics, medicine, Animals, Physical and Theoretical Chemistry, Molecular Biology, urogenital system, secretions, Organic Chemistry, Blastomere, 030104 developmental biology, lcsh:Biology (General), lcsh:QD1-999, cattle
Abstract

The bovine embryo develops in contact with the oviductal fluid (OF) during the first 4&ndash, 5 days of pregnancy. The aim of this study was to decipher the protein interactions occurring between the developing embryo and surrounding OF. In-vitro produced 4&ndash, 6 cell and morula embryos were incubated or not (controls) in post-ovulatory OF (OF-treated embryos) and proteins were then analyzed and quantified by high resolution mass spectrometry (MS) in both embryo groups and in OF. A comparative analysis of MS data allowed the identification and quantification of 56 embryo-interacting proteins originated from the OF, including oviductin (OVGP1) and several annexins (ANXA1, ANXA2, ANXA4) as the most abundant ones. Some embryo-interacting proteins were developmental stage-specific, showing a modulating role of the embryo in protein interactions. Three interacting proteins (OVGP1, ANXA1 and PYGL) were immunolocalized in the perivitelline space and in blastomeres, showing that OF proteins were able to cross the zona pellucida and be taken up by the embryo. Interacting proteins were involved in a wide range of functions, among which metabolism and cellular processes were predominant. This study identified for the first time a high number of oviductal embryo-interacting proteins, paving the way for further targeted studies of proteins potentially involved in the establishment of pregnancy in cattle.

Published
2020

42. Effect of environmental exposure to a maternally-learned odorant on anxiety-like behaviors at weaning in mice

Author

Aurélie Dewaele, Christine Baly, Vincent Bombail, Karine Badonnel, Didier Durieux, Angélique Favreau-Peigné, Marie-Annick Persuy, Biologie de la Reproduction, Environnement, Epigénétique & Développement (BREED), École nationale vétérinaire d'Alfort (ENVA)-Université de Versailles Saint-Quentin-en-Yvelines (UVSQ)-Université Paris-Saclay-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Neurobiologie de l'olfaction (NBO), Institut National de la Recherche Agronomique (INRA), Physiologie de la Nutrition et du Comportement Alimentaire (PNCA (UMR 0914)), and AgroParisTech-Université Paris-Saclay-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)

Subjects
0106 biological sciences, Offspring, [SDV]Life Sciences [q-bio], Physiology, Experimental and Cognitive Psychology, Olfaction, Weaning, Biology, Anxiety, Perinatal, 010603 evolutionary biology, 01 natural sciences, Open field, Mice, Pregnancy, Lactation, medicine, Animals, 0501 psychology and cognitive sciences, 050102 behavioral science & comparative psychology, Ecology, Evolution, Behavior and Systematics, Perinatal Exposure, Behavior, Animal, Maternal Deprivation, [SCCO.NEUR]Cognitive science/Neuroscience, 05 social sciences, Anxiety-like behavior, Environmental exposure, Environmental Exposure, medicine.disease, Open field test, medicine.anatomical_structure, Odorant, Odorants, Female, psychological phenomena and processes
Abstract

International audience; Early sensory experience, such as exposure to maternal or other environmental factors, is considered to influence neurocognitive development and behaviors. In many species, exposure to odorants during pregnancy or lactation impacts the morpho-functional development of the olfactory circuitry with changes in olfactory sensitivity, feeding behavior and food preferences at birth or later. However, few studies have investigated the impact of a perinatal exposure to odorants on the anxiety-like behavior of animals to stressfull stimuli. Here, we exposed mice to heptaldehyde (HEP) during pregnancy and lactation and measured the anxiety-like behavior of their offspring to stress-inducing novel stimuli at weaning in presence or absence of odorants. We applied a combined social and maternal separation as a stressor and measured the anxiety-like behavior in an open field (OF) in presence of two odorants, HEP or alpha-pinene (AP) as a control odorant. Although the presence of the odorant during the social separation did not influence anxiety-like behavior, we found that, if mice born to non-odorized mothers exhibited a decreased exploratory behavior in the presence of both odorants, the effect was restricted to AP for the mice perinatally exposed to HEP. These results show that anxiety-like behaviors during a stress-inducing event could be reduced by the presence of a familiar odorant. We propose that the recall of an early olfactory experience could contribute to the improvement of animal welfare in various situations associated with husbandry practices.

Published
2020

43. Effects of micronutrient supplementation on performance and epigenetic status in dairy cows

Author

Hélène Jammes, E. Mariani, J. Zawadzki, Hélène Kiefer, Marion Boutinaud, M. Fargetton, Maxime Gasselin, P. Debournoux, Audrey Prézelin, Biologie de la Reproduction, Environnement, Epigénétique & Développement (BREED), École nationale vétérinaire d'Alfort (ENVA)-Université de Versailles Saint-Quentin-en-Yvelines (UVSQ)-Université Paris-Saclay-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), École nationale vétérinaire d'Alfort (ENVA), Physiologie, Environnement et Génétique pour l'Animal et les Systèmes d'Elevage [Rennes] (PEGASE), AGROCAMPUS OUEST, Institut national d'enseignement supérieur pour l'agriculture, l'alimentation et l'environnement (Institut Agro)-Institut national d'enseignement supérieur pour l'agriculture, l'alimentation et l'environnement (Institut Agro)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), XR Reprod, Partenaires INRAE, Groupe Pilardière, and Université Paris-Saclay-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-École nationale vétérinaire d'Alfort (ENVA)-Université de Versailles Saint-Quentin-en-Yvelines (UVSQ)

Subjects
040301 veterinary sciences, Soybean meal, Ice calving, Biology, methylome, SF1-1100, Epigenesis, Genetic, 0403 veterinary science, 03 medical and health sciences, cows, Animal science, Casein, Lactation, medicine, Animals, Metritis, 030304 developmental biology, 2. Zero hunger, 0303 health sciences, [SDV.BA]Life Sciences [q-bio]/Animal biology, Postpartum Period, food and beverages, 04 agricultural and veterinary sciences, medicine.disease, Micronutrient, Mastitis, Diet, Animal culture, medicine.anatomical_structure, Milk, micronutrients, [SDV.SA.SPA]Life Sciences [q-bio]/Agricultural sciences/Animal production studies, Dietary Supplements, Leukocytes, Mononuclear, Animal Science and Zoology, Cattle, Female, peripartum, performances, Postpartum period
Abstract

International audience; The postpartum period is crucial in dairy cows and is marked by major physiological and metabolic changes that affect milk production, immune response and fertility. Nutrition remains the most important lever for limiting the negative energy balance and its consequences on general health status in highly selected dairy cows. In order to analyze the effect of a commercial micronutrient on intrinsic parameters, performances and the epigenome of dairy cows, 2 groups of 12 Holstein cows were used: 1 fed a standard diet (mainly composed of corn silage, soybean meal and non-mineral supplement) and the other 1 fed the same diet supplemented with the commercial micronutrient (µ-nutrient supplementation) for 4 weeks before calving and 8 weeks thereafter. Milk production and composition, BW, body condition score (BCS), DM intake (DMI) and health (calving score, metritis and mastitis) were recorded over the study period. Milk samples were collected on D15 and D60 post-calving for analyses of casein, Na+ and K+ contents and metalloprotease activity. Milk leukocytes and milk mammary epithelial cells (mMECs) were purified and counted. The viability of mMECs was assessed, together with their activity, through an analysis of gene expression. At the same time points, peripheral blood mononuclear cells (PBMCs) were purified and counted. Using genomic DNA extracted from PBMCs, mMECs and milk leukocytes, we assessed global DNA methylation (Me-CCGG) to evaluate the epigenetic imprinting associated with the µ-nutrient-supplemented diet. The µ-nutrient supplementation increased BCS and BW without modifying DMI or milk yield and composition. It also improved calving condition, reducing the time interval between calving and first service. Each easily collectable cell type displayed a specific pattern of Me-CCGG with only subtle changes associated with lactation stages in PBMCs. In conclusion, the response to the µ-nutrient supplementation improved the body condition without alteration of global epigenetic status in dairy cows.

Published
2020

44. Prenatal air pollution exposure to diesel exhaust induces cardiometabolic disorders in adulthood in a sex-specific manner

Author

Rousseau-Ralliard, Delphine, Richard, Christophe, Hoarau, Pauline, Lallemand, Marie-Sylvie, Morillon, Lucie, Aubrière, Marie-Christine, Valentino, Sarah A, Dahirel, Michèle, Guinot, Marine, Fournier, Natalie, Morin, Gwendoline, Mourier, Eve, Camous, Sylvaine, Slama, Rémy, Cassee, Flemming R, Couturier-Tarrade, Anne, Chavatte-Palmer, Pascale, IRAS OH Toxicology, dIRAS RA-1, IRAS OH Toxicology, dIRAS RA-1, Biologie de la Reproduction, Environnement, Epigénétique & Développement (BREED), Université Paris-Saclay-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-École nationale vétérinaire d'Alfort (ENVA)-Université de Versailles Saint-Quentin-en-Yvelines (UVSQ), Lipides : systèmes analytiques et biologiques (Lip(Sys)²), Université Paris-Saclay, Sciences de l'Animal et de l'Aliment de Jouy (SAAJ), Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Institute for Advanced Biosciences / Institut pour l'Avancée des Biosciences (Grenoble) (IAB), Centre Hospitalier Universitaire [Grenoble] (CHU)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Etablissement français du sang - Auvergne-Rhône-Alpes (EFS)-Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes (UGA), Utrecht University [Utrecht], Chavatte-Palmer, Pascale, and École nationale vétérinaire - Alfort (ENVA)-Université de Versailles Saint-Quentin-en-Yvelines (UVSQ)-Université Paris-Saclay-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)

Subjects
Adult, Male, Offspring, [SDV]Life Sciences [q-bio], Birth weight, Gestational exposure, Physiology, Biochemistry, 03 medical and health sciences, 0302 clinical medicine, Pregnancy, Environmental Science(all), Air Pollution, 11. Sustainability, Fetal programming, medicine, Animals, Humans, Weaning, 030212 general & internal medicine, Vehicle Emissions, 030304 developmental biology, General Environmental Science, 2. Zero hunger, Intergenerational effect, 0303 health sciences, Inhalation, business.industry, Adult offspring, diesel exhaust, medicine.disease, 3. Good health, [SDV] Life Sciences [q-bio], Blood pressure, Rabbit biomedical model, [SDV.SPEE] Life Sciences [q-bio]/Santé publique et épidémiologie, Cardiovascular Diseases, Maternal Exposure, 13. Climate action, In utero, Prenatal Exposure Delayed Effects, Gestation, [SDV.SPEE]Life Sciences [q-bio]/Santé publique et épidémiologie, Female, Rabbits, Metabolic syndrome, business
Abstract

Background Results from observational and experimental studies indicate that exposure to air pollutants during gestation reduces birth weight, whereas little is known on potential cardiometabolic consequences for the offspring at adulthood. Objectives Our aim was to evaluate the long-term effects of gestational exposure to diesel engine exhaust (DE) on adult offspring phenotype in a rabbit model. Methods The protocol was designed to mimic human exposure in large European cities. Females rabbits were exposed to diluted (1 mg/m3) DE (exposed, n = 9) or clean air (controls, n = 7), from 3 days after mating, 2 h/d and 5 d/wk in a nose-only inhalation system throughout gestation (gestation days 3–27). After birth and weaning, 72 offspring (47 exposed and 25 controls) were raised until adulthood (7.5 months) to evaluate their cardio-metabolic status, including the monitoring of body weight and food intake, fasting biochemistry, body composition (iDXA), cardiovascular parameters and glucose tolerance. After a metabolic challenge (high fat diet in males and gestation in females), animals were euthanized for postmortem phenotyping. Results Sex-specific responses to maternal exposure were observed in adult offspring. Age-related increases in blood pressure (p = 0.058), glycaemia (p = 0.029), and perirenal fat mass (p = 0.026) as well as reductions in HDL-cholesterol (p = 0.025) and fat-to-body weight ratio (p = 0.011) were observed in exposed males, suggesting a metabolic syndrome. Almost only trends were observed in exposed females with higher triglycerides and decreased bone density compared to control females. Metabolic challenges triggered or amplified some biological responses, especially in females. Conclusions In utero exposure to air pollution predisposed rabbit offspring to cardiometabolic disorders in a sex-specific manner.

Published
2021

45. Genetic merit for fertility alters the bovine uterine luminal fluid proteome†

Author

Georg J. Arnold, Gilles Charpigny, Miwako Kösters, Thomas Fröhlich, Katrin Gegenfurtner, Pascal Mermillod, Olivier Desnoës, Patrick Lonergan, Olivier Sandra, Pascal Salvetti, Sébastien Fritz, Florian Flenkenthaler, Daniel Le Bourhis, Eckhard Wolf, Laboratory for Functional Genome Analysis (LAFUGA), Ludwig-Maximilians-Universität München (LMU), Génétique Animale et Biologie Intégrative (GABI), Université Paris-Saclay-AgroParisTech-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Station de Phénotypage, Allice, Union Evolution, Partenaires INRAE, Biologie de la Reproduction, Environnement, Epigénétique & Développement (BREED), École nationale vétérinaire d'Alfort (ENVA)-Université de Versailles Saint-Quentin-en-Yvelines (UVSQ)-Université Paris-Saclay-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Physiologie de la reproduction et des comportements [Nouzilly] (PRC), Institut Français du Cheval et de l'Equitation [Saumur]-Université de Tours (UT)-Centre National de la Recherche Scientifique (CNRS)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), University College Dublin [Dublin] (UCD), European Project: 0312097(2003), AgroParisTech-Université Paris-Saclay-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), École nationale vétérinaire - Alfort (ENVA)-Université de Versailles Saint-Quentin-en-Yvelines (UVSQ)-Université Paris-Saclay-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Institut Français du Cheval et de l'Equitation [Saumur] (IFCE)-Université de Tours (UT)-Centre National de la Recherche Scientifique (CNRS)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Université Paris-Saclay-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-École nationale vétérinaire d'Alfort (ENVA)-Université de Versailles Saint-Quentin-en-Yvelines (UVSQ), and Institut Français du Cheval et de l'Equitation [Saumur]-Université de Tours-Centre National de la Recherche Scientifique (CNRS)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)

Subjects
0301 basic medicine, Proteomics, Proteome, media_common.quotation_subject, [SDV]Life Sciences [q-bio], Fertility, Biology, [SDV.BDLR.RS]Life Sciences [q-bio]/Reproductive Biology/Sexual reproduction, Andrology, 03 medical and health sciences, Tandem Mass Spectrometry, Genetic predisposition, Conceptus, Vitamin metabolic process, Animals, dairy cows, Folate Receptor 1, media_common, 2. Zero hunger, Uterus, 0402 animal and dairy science, Maternal effect, Computational Biology, 04 agricultural and veterinary sciences, Cell Biology, General Medicine, 040201 dairy & animal science, early development, Interferon tau, Lactotransferrin, Lactoferrin, 030104 developmental biology, Reproductive Medicine, uterine fluid, Chromogranin A, Cattle, Female
Abstract

Over the last decades, fertility of dairy cows has declined due to selection strategies focusing on milk yield. To study the effect of genetic merit for fertility on the proteome of the bovine uterine luminal fluid, Holstein heifers with low- and two groups of heifers with high-fertility index (high-fertility Holstein and Montbéliarde) were investigated. To focus on the maternal effect, heifers from all groups were synchronized and received on Day 7 high-quality embryos. Uterine luminal fluid from Day 19 pregnant heifers was analyzed in a holistic proteomic approach using nano-LC-MS/MS analysis combined with a label-free quantification approach. In total, 1737 proteins were identified, of which 597 differed significantly in abundance between the three groups. The vast majority of proteome differences was found comparing both high-fertility groups to the low-fertility Holstein group, showing that the genetic predisposition for fertility is prevalent regarding the uterine luminal fluid proteome. Evaluation of this dataset using bioinformatic tools revealed an assignment of higher abundant proteins in low-fertility Holstein to several metabolic processes, such as vitamin metabolic process, which comprises folate receptor alpha (FOLR1) and retinol-binding protein, indicating an involvement of disturbed metabolic processes in decreased fertility. Moreover, immune system-related proteins — lactotransferrin and chromogranin A — were enriched in low-fertility cows together with interferon tau 3 h and interferon tau-2. Our results indicate that the genetic merit for fertility leads to substantial quantitative differences at the level of proteins in uterine fluid of pregnant animals, thus altering the microenvironment for the early conceptus.

Published
2019

46. Preimplantation development in ungulates: a 'ménage à quatre' scenario

Author

Hervé Acloque, Jérôme Artus, Isabelle Hue, Inserm UMR935, AP-HP Hôpital Bicêtre (Le Kremlin-Bicêtre), Biologie de la Reproduction, Environnement, Epigénétique & Développement (BREED), École nationale vétérinaire - Alfort (ENVA)-Université de Versailles Saint-Quentin-en-Yvelines (UVSQ)-Université Paris-Saclay-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Génétique Animale et Biologie Intégrative (GABI), AgroParisTech-Université Paris-Saclay-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Acloque, Hervé, École nationale vétérinaire d'Alfort (ENVA)-Université de Versailles Saint-Quentin-en-Yvelines (UVSQ)-Université Paris-Saclay-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), and Université Paris-Saclay-AgroParisTech-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)

Subjects
0301 basic medicine, Embryology, Embryonic Development, Biology, [SDV.BDLR.RS]Life Sciences [q-bio]/Reproductive Biology/Sexual reproduction, 03 medical and health sciences, 0302 clinical medicine, Endocrinology, Embryonic disc, medicine, Inner cell mass, Conceptus, Animals, Blastocyst, Zona pellucida, Perissodactyla, Artiodactyla, 030219 obstetrics & reproductive medicine, [SDV.BDLR.RS] Life Sciences [q-bio]/Reproductive Biology/Sexual reproduction, Embryogenesis, Uterus, Obstetrics and Gynecology, Trophoblast, [SDV.BDD.EO] Life Sciences [q-bio]/Development Biology/Embryology and Organogenesis, Cell Biology, Embryonic stem cell, Cell biology, 030104 developmental biology, medicine.anatomical_structure, [SDV.BDD.EO]Life Sciences [q-bio]/Development Biology/Embryology and Organogenesis, Reproductive Medicine, embryonic structures, Female
Abstract

International audience; In ungulates, early embryonic development differs dramatically from that of mice and humans and is characterized by an extended period of pre-and peri-implantation development in utero. After hatching from the zona pellucida, the ungulate blastocyst will stay free in the uterus for many days before implanting within the uterine wall. During this protracted peri-implantation period, an intimate dialog between the embryo and the uterus is established through a complex series of paracrine signals. The blastocyst elongates, leading to extreme growth of extra-embryonic tissues, and at the same time, the inner cell mass moves up into the trophoblast and evolves into the embryonic disc, which is directly exposed to molecules present in the uterine fluids. In the peri-implantation period, uterine glands secrete a wide range of molecules, including enzymes, growth factors, adhesion proteins, cytokines, hormones, and nutrients like amino and fatty acids, which are collectively referred to as histotroph. The identification, role, and effects of these secretions on the biology of the conceptus are still being described; however, the studies that have been conducted to date have demonstrated that histotroph is essential for embryonic development and serves a critical function during the pre-and peri implantation periods. Here, we present an overview of current knowledge on the molecular dialogue among embryonic, extraembryonic, and maternal tissues prior to implantation. Taken together, the body of work described here demonstrates the extent to which this dialog enables the coordination of the development of the conceptus with respect to the establishment of embryonic and extra-embryonic tissues as well as in preparation for implantation. Reproduction (2020) 159 R151-R172

Published
2019

47. No-Contact Microchip Monitoring of Body Temperature in Yearling Horses

Author

Marie Frejaville, Tristan Jousset, Florence Jaffrezic, Laurence Wimel, Pascale Chavatte-Palmer, Juliette Auclair-Ronzaud, Cédric Dubois, Stéphanie Benoist, Institut Français du Cheval et de l'Equitation, Génétique Animale et Biologie Intégrative (GABI), AgroParisTech-Université Paris-Saclay-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Université Paris-Saclay, Biologie de la Reproduction, Environnement, Epigénétique & Développement (BREED), Université Paris-Saclay-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-École nationale vétérinaire d'Alfort (ENVA)-Université de Versailles Saint-Quentin-en-Yvelines (UVSQ), Institut Français du Cheval et de l'Equitation [Saumur], Université Paris-Saclay-AgroParisTech-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), and École nationale vétérinaire d'Alfort (ENVA)-Université de Versailles Saint-Quentin-en-Yvelines (UVSQ)-Université Paris-Saclay-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)

Subjects
Male, 040301 veterinary sciences, [SDV]Life Sciences [q-bio], Drinking, Forage, Biology, Body Temperature, 0403 veterinary science, Animal science, Animals, [INFO]Computer Science [cs], Horses, equine, [SDV.GEN]Life Sciences [q-bio]/Genetics, [SDV.BA.MVSA]Life Sciences [q-bio]/Animal biology/Veterinary medicine and animal Health, 0402 animal and dairy science, Free access, Temperature, Horse, Rectal temperature, microchip, 04 agricultural and veterinary sciences, Prostheses and Implants, 040201 dairy & animal science, monitoring, [SDV.GEN.GA]Life Sciences [q-bio]/Genetics/Animal genetics, sexual dimorphism, Female, Seasons
Abstract

International audience; In clinics, temperature is used as an indicator of health. Mostly rectal temperature is recorded, requiring handling and time. Temperature-sensitive identification microchips could be an alternative. Foals (26 males and 17 females), 4-12 months old, were housed in stalls over two winters (December-February). They were equipped with an identification and temperature sensor microchip implanted in the neckline. Temperature was recorded using an antenna located near the drinking trough. Animals were fed concentrated feed and forage twice daily, with free access to water. Rectal temperatures (79 measurements) were recorded simultaneously in 26 animals. Data were analyzed with a linear mixed model, using natural cubic splines for the mean curve and a random horse effect. All animals remained healthy throughout the study. More than 100,000 recordings were obtained. Mean temperature for all individuals at all times was 37.5 +/- 0.1 degrees C. Time of the day affected temperature with a daily amplitude of 0.96 degrees C (P < .001). Lowest temperatures were observed before dawn, the acrophase occurring around 18:00, with a smaller increase around midday. Mean temperature was 0.26 degrees C higher in males (P < .05). It was also 0.1 degrees C higher in light (

Published
2019

48. DNA methylation stability in fish spermatozoa upon external constraint: Impact of fish hormonal stimulation and sperm cryopreservation

Author

Hélène Jammes, Pierre-Yves Le Bail, Alexandra Depince, Anne Gabory, Catherine Labbé, Katarzyna Dziewulska, Laboratoire de Physiologie et Génomique des Poissons (LPGP), Structure Fédérative de Recherche en Biologie et Santé de Rennes ( Biosit : Biologie - Santé - Innovation Technologique )-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Biologie de la Reproduction, Environnement, Epigénétique & Développement (BREED), École nationale vétérinaire d'Alfort (ENVA)-Université de Versailles Saint-Quentin-en-Yvelines (UVSQ)-Université Paris-Saclay-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Agricultural University of Szczecin, Partenaires INRAE, ANR-11-INBS-0003,CRB-Anim,Réseau de Centres de Ressources Biologiques pour les animaux domestiques(2011), European Project: COST Action FA 1205, AQUAGAMETE, European Project: COST Action FA1201,EPICONCEPT, École nationale vétérinaire - Alfort (ENVA)-Université de Versailles Saint-Quentin-en-Yvelines (UVSQ)-Université Paris-Saclay-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), and ANR‐11‐INBS‐0003, Agence Nationale de la Recherche

Subjects
0301 basic medicine, Male, endocrine system, [SDV]Life Sciences [q-bio], gamete, Fertilization in Vitro, Biology, Cryopreservation, Andrology, Gonadotropin-Releasing Hormone, 03 medical and health sciences, 0302 clinical medicine, Cryoprotective Agents, Goldfish, Genetics, medicine, Animals, [INFO]Computer Science [cs], Dimethyl Sulfoxide, Epigenetics, Zebrafish, 030219 obstetrics & reproductive medicine, urogenital system, Methanol, Embryo, Cell Biology, Methylation, DNA Methylation, Sperm, Propylene Glycol, Spermatozoa, Domperidone, Drug Combinations, 030104 developmental biology, medicine.anatomical_structure, DNA methylation, Oocytes, Sperm Motility, Gamete, Female, Spermatogenesis, epigenetic, Developmental Biology, biotechnology, Semen Preservation
Abstract

International audience; Highly differentiated mature spermatozoa carry not only genetic but also epigenetic information that is to be transmitted to the embryo. DNA methylation is one epigenetic actor associated with sperm nucleus compaction, gene silencing, and prepatterning of embryonic gene expression. Therefore, the stability of this mark toward reproductive biotechnologies is a major issue in animal production. The present work explored the impact of hormonal induction of spermiation and sperm cryopreservation in two cyprinids, the goldfish (Carassius auratus) and the zebrafish (Danio rerio), using LUminometric Methylation Assay (LUMA). We showed that while goldfish hormonal treatment did increase sperm production, it did not alter global DNA methylation of spermatozoa. Different sperm samples repeatedly collected from the same males for 2 months also showed the same global DNA methylation level. Similarly, global DNA methylation was not affected after cryopreservation of goldfish spermatozoa with methanol, whereas less efficient cryoprotectants (dimethylsulfoxide and 1,2‐propanediol) decreased DNA methylation. In contrast, cryopreservation of zebrafish spermatozoa with methanol induced a slight, but significant, increase in global DNA methylation. In the less compact nuclei, that is, goldfish fin somatic cells, cryopreservation did not change global DNA methylation regardless of the choice of cryoprotectant. To conclude, global DNA methylation is a robust parameter with respect to biotechnologies such as hormonal induction of spermiation and sperm cryopreservation, but it can be altered when the best sperm manipulation conditions are not met.

Published
2019

49. Oocyte maturation under lipotoxic conditions induces carryover transcriptomic and functional alterations during post-hatching development of good-quality blastocysts: novel insights from a bovine embryo-transfer model

Author

Ken Declerck, Pieter Meysman, Wim Vanden Berghe, Christophe Richard, Katrien Sprangers, Waleed F.A. Marei, K. L. J. Desmet, Peter E.J. Bols, Isabelle Hue, Gerrit T.S. Beemster, Jo L.M.R. Leroy, Kris Laukens, Department of Veterinary Sciences, University of Antwerp (UA), Department of Theriogenology, Cairo University, Biologie de la Reproduction, Environnement, Epigénétique & Développement (BREED), École nationale vétérinaire d'Alfort (ENVA)-Université de Versailles Saint-Quentin-en-Yvelines (UVSQ)-Université Paris-Saclay-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Department of Biology, Department of Mathematics & Computer Science (Antwerp), Department of Biomedical Sciences, and This study was supported by the Flemish Research Fund (FWO grant 11L8716N and FWO project 42/FAO10300/6541)

Subjects
animal structures, Veterinary medicine, interferon-τ, embryo, Biology, fatty acids, maternal metabolism, Transcriptome, Andrology, 03 medical and health sciences, 0302 clinical medicine, Pregnancy, Gene expression, medicine, metabolic disorders, Animals, Humans, implantation, Epigenetics, 030304 developmental biology, 2. Zero hunger, 0303 health sciences, 030219 obstetrics & reproductive medicine, embryo metabolism, Rehabilitation, Embryogenesis, Obstetrics and Gynecology, Embryo, Embryo culture, [SDV.BDLR]Life Sciences [q-bio]/Reproductive Biology, Oocyte, Embryo Transfer, Embryo transfer, In Vitro Oocyte Maturation Techniques, medicine.anatomical_structure, Blastocyst, Reproductive Medicine, embryonic structures, gene expression, Oocytes, Cattle, Female, Human medicine
Abstract

STUDY QUESTION Does oocyte maturation under lipolytic conditions have detrimental carry-over effects on post-hatching embryo development of good-quality blastocysts after transfer? SUMMARY ANSWER Surviving, morphologically normal blastocysts derived from bovine oocytes that matured under lipotoxic conditions exhibit long-lasting cellular dysfunction at the transcriptomic and metabolic levels, which coincides with retarded post-hatching embryo development. WHAT IS KNOWN ALREADY There is increasing evidence showing that following maturation in pathophysiologically relevant lipotoxic conditions (as in obesity or metabolic syndrome), surviving blastocysts of good (transferable) morphological quality have persistent transcriptomic and epigenetic alteration even when in vitro embryo culture takes place under standard conditions. However, very little is known about subsequent development in the uterus after transfer. STUDY DESIGN, SIZE, DURATION Bovine oocytes were matured in vitro in the presence of pathophysiologically relevant, high non-esterified fatty acid (NEFA) concentrations (HIGH PA), or in basal NEFA concentrations (BASAL) as a physiological control. Eight healthy multiparous non-lactating Holstein cows were used for embryo transfers. Good-quality blastocysts (pools of eight) were transferred per cow, and cows were crossed over for treatments in the next replicate. Embryos were recovered 7 days later and assessed for post-hatching development, phenotypic features and gene expression profile. Blastocysts from solvent-free and NEFA-free maturation (CONTROL) were also tested for comparison. PARTICIPANTS/MATERIALS, SETTING, METHODS Recovered Day 14 embryos were morphologically assessed and dissected into embryonic disk (ED) and extraembryonic tissue (EXT). Samples of EXT were cultured for 24 h to assess cellular metabolic activity (glucose and pyruvate consumption and lactate production) and embryos’ ability to signal for maternal recognition of pregnancy (interferon-τ secretion; IFN-τ). ED and EXT samples were subjected to RNA sequencing to evaluate the genome-wide transcriptome patterns. MAIN RESULTS AND THE ROLE OF CHANCE The embryo recovery rate at Day 14 p.i. was not significantly different among treatment groups (P > 0.1). However, higher proportions of HIGH PA embryos were retarded in growth (in spherical stage) compared to the more elongated tubular stage embryos in the BASAL group (P LARGE-SCALE DATA The datasets of RNA sequencing analysis are available in the NCBI’s Gene Expression Omnibus (GEO) repository, series accession number GSE127889 (https://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE127889). Datasets of differentially expressed genes and their gene ontology functions are available in the Mendeley datasets at http://dx.doi.org/10.17632/my2z7dvk9j.2. LIMITATIONS, REASONS FOR CAUTION The bovine model was used here to allow non-invasive embryo transfer and post-hatching recovery on Day 14. There are physiological differences in some characteristics of post-hatching embryo development between human and cows, such as embryo elongation and trophoblastic invasion. However, the main carry-over effects of oocyte maturation under lipolytic conditions described here are evident at the cellular level and therefore may also occur during post-hatching development in other species including humans. In addition, post-hatching development was studied here under a healthy uterine environment to focus on carry-over effects originating from the oocyte, whereas additional detrimental effects may be induced by maternal metabolic disorders due to adverse changes in the uterine microenvironment. RNA sequencing results were not verified by qPCR, and no solvent control was included. WIDER IMPLICATIONS OF THE FINDINGS Our observations may increase the awareness of the importance of maternal metabolic stress at the level of the preovulatory oocyte in relation to carry-over effects that may persist in the transferrable embryos. It should further stimulate new research about preventive and protective strategies to optimize maternal metabolic health around conception to maximize embryo viability and thus fertility outcome. STUDY FUNDING/COMPETING INTEREST(S) This study was supported by the Flemish Research Fund (FWO grant 11L8716N and FWO project 42/FAO10300/6541). The authors declare there are no conflicts of interest.

Published
2019

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