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12. Effects of cholesterol content on activity of P-glycoproteins and membrane physical state, and consequences for anthelmintic resistance in the nematode Haemonchus contortus

Author

Riou Mickaël, Guégnard Fabrice, Le Vern Yves, Grasseau Isabelle, Koch Christine, Blesbois Elisabeth, and Kerboeuf Dominique

Subjects
parasite, eggshell, mβcd, fluidity, cholesterol, p-glycoproteins, resistance, anthelmintics, Infectious and parasitic diseases, RC109-216
Abstract

Eukaryote plasma membranes protect cells from chemical attack. Xenobiotics, taken up through passive diffusion, accumulate in the membranes, where they are captured by transporters, among which P-glycoproteins (Pgps). In nematodes such as Haemonchus contortus, eggshells and cuticles provide additional protective barriers against xenobiotics. Little is known about the role of these structures in the transport of chemical molecules. Pgps, members of the ABC transporter family, are present in eggshells and cuticles. Changes in the activity of these proteins have also been correlated with alterations in lipids, such as cholesterol content, in eggshells. However, the cellular mechanisms underlying these effects remain unclear. We show here that an experimental decrease in the cholesterol content of eggshells of Haemonchus contortus, with Methyl-beta-CycloDextrin (MβCD), results in an increase in membrane fluidity, favouring Pgp activity and leading to an increase in resistance to anthelmintics. This effect is modulated by the initial degree of anthelminthic resistance of the eggs. These results suggest that eggshell fluidity plays a major role in the modulation of Pgp activity. They confirm that Pgp activity is highly influenced by the local microenvironment, in particular sterols, as observed in some vertebrate models. Thus, eggshell barriers could play an active role in the transport of xenobiotics.

Published
2020
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14. Impact of glycerol on sperm fertilizing capacity in chicken

Author

Lin, Hsiu-Lien, Grasseau, Isabelle, Mermillod, Pascal, Blesbois, Elisabeth, Vitorino Carvalho, Anais, and Vitorino Carvalho, Anaïs

Subjects
[SDV.BA] Life Sciences [q-bio]/Animal biology, [SDV.BDD] Life Sciences [q-bio]/Development Biology, [SDV.BBM] Life Sciences [q-bio]/Biochemistry, Molecular Biology, [SDV.BC] Life Sciences [q-bio]/Cellular Biology, [SDV.BDLR] Life Sciences [q-bio]/Reproductive Biology, [SDV.BIO] Life Sciences [q-bio]/Biotechnology
Published
2022

15. Application of colloidal centrifugation to remove glycerol from chicken frozen-thawed semen

Author

Lin, Hsiu-Lien, Grasseau, Isabelle, Mermillod, Pascal, Blesbois, Elisabeth, Vitorino Carvalho, Anais, and Vitorino Carvalho, Anaïs

Subjects
[SDV.BA] Life Sciences [q-bio]/Animal biology, [SDV.BDD] Life Sciences [q-bio]/Development Biology, [SDV.BBM] Life Sciences [q-bio]/Biochemistry, Molecular Biology, [SDV.BC] Life Sciences [q-bio]/Cellular Biology, [SDV.BDLR] Life Sciences [q-bio]/Reproductive Biology, [SDV.BIO] Life Sciences [q-bio]/Biotechnology
Published
2022

16. Elimination du glycérol de la semence décongelée de coq par centrifugation dans un gel colloïdal synthétique

Author

Lin, Hsiu-Lien, Grasseau, Isabelle, Mermillod, Pascal, Blesbois, Elisabeth, Vitorino Carvalho, Anais, Vitorino Carvalho, Anaïs, and Infrastructures - Réseau de Centres de Ressources Biologiques pour les animaux domestiques - - CRB-Anim2011 - ANR-11-INBS-0003 - INBS - VALID

Subjects
[SDV.BA] Life Sciences [q-bio]/Animal biology, [SDV.BDD] Life Sciences [q-bio]/Development Biology, [SDV.BBM] Life Sciences [q-bio]/Biochemistry, Molecular Biology, [SDV.BC] Life Sciences [q-bio]/Cellular Biology, [SDV.BDLR] Life Sciences [q-bio]/Reproductive Biology, [SDV.BIO] Life Sciences [q-bio]/Biotechnology
Published
2022

21. Nouvelles avancées dans la cryoconservation des spermatozoïdes de coq

Author

Lin, Hsiu-Lien, Grasseau, Isabelle, Mermillod, Pascal, Blesbois, Elisabeth, Vitorino Carvalho, Anais, and Vitorino Carvalho, Anaïs

Subjects
[SDV.BA] Life Sciences [q-bio]/Animal biology, [SDV.BDD] Life Sciences [q-bio]/Development Biology, [SDV.BBM] Life Sciences [q-bio]/Biochemistry, Molecular Biology, [SDV.BC] Life Sciences [q-bio]/Cellular Biology, [SDV.BDLR] Life Sciences [q-bio]/Reproductive Biology, [SDV.BIO] Life Sciences [q-bio]/Biotechnology
Published
2021

25. Seminal plasma exosomes of fertile and subfertile roosters have morphological and biochemical characteristics that affect sperm viability

Author

Cordeiro, Luiz, Vitorino Carvalho, Anais, Grasseau, Isabelle, Lin, Hsiu-Lien, Usbekov, R., Blesbois, Elisabeth, Physiologie de la reproduction et des comportements [Nouzilly] (PRC), 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), 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), France The french branch of WPSA (WPSA). FRA., 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 ProdInra, Archive Ouverte

Subjects
[SDV] Life Sciences [q-bio], urogenital system, [SDV]Life Sciences [q-bio]
Abstract

Seminal plasma exosomes of fertile and subfertile roosters have morphological and biochemical characteristics that affect sperm viability. World poultry congress WPC 2020

Published
2020

26. Exploration of the spermatozoa functions involved in the infertility of broiler infertility

Author

Vitorino Carvalho, Anais, Thelie, Aurore, Soler-Vasco, Laura, Cordeiro, Luiz, Grasseau, Isabelle, Labas, Valérie, Teixeira-Gomes, Ana Paula, Blesbois, Elisabeth, Physiologie de la reproduction et des comportements [Nouzilly] (PRC), Centre National de la Recherche Scientifique (CNRS)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Université de Tours-Institut Français du Cheval et de l'Equitation [Saumur], Biosynthèse & Toxicité des Mycotoxines (ToxAlim-BioToMyc), ToxAlim (ToxAlim), Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Institut National Polytechnique (Toulouse) (Toulouse INP), Université Fédérale Toulouse Midi-Pyrénées-Ecole Nationale Vétérinaire de Toulouse (ENVT), Institut National Polytechnique (Toulouse) (Toulouse INP), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Ecole d'Ingénieurs de Purpan (INPT - EI Purpan), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Institut National de la Recherche Agronomique (INRA)-Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Institut National de la Recherche Agronomique (INRA), Infectiologie et Santé Publique (UMR ISP), Université de Tours-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), Université de Tours (UT)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), 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), Infectiologie Animale et Santé Publique - IASP (Nouzilly, France), and Institut National de la Recherche Agronomique (INRA)

Subjects
[SDV]Life Sciences [q-bio]
Abstract

International audience; Exploration of the spermatozoa functions involved in the infertility of broiler infertility. 19th International Congress on Animal Reproduction ICAR 2020

Published
2020

28. Chicken semen cryopreservation: importance of cryoprotectants.

Author

Lin, Hsiu-Lien Herbie, Blesbois, Elisabeth, and Vitorino Carvalho, Anaïs

Abstract

Sperm cryopreservation is an important strategy to conserve animal genetic diversity and transmit superior genetic background, facilitated by a non-invasive sampling and collection of large amounts of sperm. In chickens, various freezing protocols and cryoprotectants (CPAs) have been developed to freeze sperm, but each lab still debates and competes to disclose the most suitable freezing conditions. Many key points, such as semen donor selection, collection tips, diluent compositions, CPA selection, pre-freezing manipulation, semen packaging type, freezing and thawing rates, are all impactful to the efficiency of chicken sperm cryopreservation. Whereas glycerol is believed as the most efficient and the less toxic CPA, it also presents an unwanted contraceptive effect after insemination. Thus, one of the major developed strategies to conquer this issue is removing glycerol before insemination, but current protocols remain not efficient to completely abolish glycerol contraceptive effect. Novel methodologies will bring us a better understanding of the cellular and molecular mechanisms involved in sperm cryobiology to decipher the challenges of chicken sperm cryopreservation. [ABSTRACT FROM AUTHOR]

Published
2022
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34. Pregnancy after short exposure of cryopreserved porcine embryo to cryoprotective agents

Author

Guignot, Florence, Dubuisson, Florine, Camille, Voiry, Eric, Royer, Anais, Giovanetti, Christian, Moussu, Frédéric, Elleboudt, jonathan, savoie, Woelders, Henri, Mermillod, Pascal, Blesbois, Elisabeth, Physiologie de la reproduction et des comportements [Nouzilly] (PRC), Institut National de la Recherche Agronomique (INRA)-Institut Français du Cheval et de l'Equitation [Saumur]-Université de Tours (UT)-Centre National de la Recherche Scientifique (CNRS), Unité Expérimentale de Physiologie Animale de l‘Orfrasiére (UE PAO), Institut National de la Recherche Agronomique (INRA), Wageningen UR Livestock Research, Guignot, Florence, and Institut National de la Recherche Agronomique (INRA)-Institut Français du Cheval et de l'Equitation [Saumur]-Université de Tours-Centre National de la Recherche Scientifique (CNRS)

Subjects
[SDV] Life Sciences [q-bio], [SDV]Life Sciences [q-bio], ComputingMilieux_MISCELLANEOUS
Abstract

International audience

Published
2019

35. Breakthrough to improve the reproductive capacity of gene bank material

Author

Blesbois, Elisabeth, LIPTOI, K., Woelders, Henri, B, berna-juarez, S, Nandi, Thelie, Aurore, govoroun, marina, E, varkonyi, Guignot, Florence, Santiago-Moreno, Julian, Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), and Guignot, Florence

Subjects
[SDV] Life Sciences [q-bio], [SDV]Life Sciences [q-bio], ComputingMilieux_MISCELLANEOUS
Abstract

International audience

Published
2019

36. The seminal acrosin‐inhibitor ClTI1/SPINK2 is a fertility‐associated marker in the chicken

Author

Thélie, Aurore and Blesbois, Elisabeth

Subjects
fertility, proteomics, chicken, seminal plasma
Abstract

A paper produced in the frame of the H2020 project 677353 'Innovative Management of Animal GEnetic resources' This study investigates the role of components of the seminal plasma on malefertility and identifies SPINK2 protein as an important actor of fertility in birds through its innibitory action on acrosin isoforms proteins.&nbsp

Published
2019

37. Seminal plasma amino acid profile in different breeds of chicken: Role of seminal plasma on sperm cryoresistance

Author

Santiago-Morenos, Julian and Blesbois, Elisabeth

Subjects
chicken, frozen semen, seminal plasma, breed variability
Abstract

A paper produced in the frame of the H2020 project 677353 'Innovative Management of Animal GEnetic resources' This paper reveals the role of the seminal plasma composition in the differencesbetween chicken breeds regarding fertility obtained from frozen semen. Finally, removing seminal plasma decreases the variability of results and damages to DNA (fragmentation).

Published
2019

38. How vitrification and in vitro culture affect pig embryos? Clues from RNA-sequencing data

Author

Alminana, Carmen, Bauersachs, Stefan, Blesbois, Elisabeth, Mermillod, Pascal, Guignot, Florence, Guignot, Florence, and Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)

Subjects
[SDV] Life Sciences [q-bio], [SDV]Life Sciences [q-bio], ComputingMilieux_MISCELLANEOUS
Abstract

International audience

Published
2019

39. Chicken semen cryopreservation and use for the restoration of rare genetic resources

Author

Thelie, Aurore and Blesbois, Elisabeth

Subjects
fertility, genetic resources, chicken, cryopreservation
Abstract

A paper produced in the frame of the H2020 project 677353 'Innovative Management of Animal GEnetic resources' This paper analyses a practical case of using a semen bank to recoverlost families from a chicken experimental line. It provides informations to optimize the process and highlights the importance of standardized procedures to improve the performance.

Published
2019

40. Seminal plasma amino acid profile in different breeds of chicken : Role of seminal plasma on sperm cryoresistance

Author

Santiago-Moreno, Julián, Bernal, Berenice, Pérez-Cerezales, Serafín, Castaño, Cristina, Toledano-Díaz, Adolfo, Esteso, Milagros C., Gutiérrez-Adán, Alfonso, López-Sebastián, Antonio, Gil, María G., Woelders, Henri, Blesbois, Elisabeth, Santiago-Moreno, Julián, Bernal, Berenice, Pérez-Cerezales, Serafín, Castaño, Cristina, Toledano-Díaz, Adolfo, Esteso, Milagros C., Gutiérrez-Adán, Alfonso, López-Sebastián, Antonio, Gil, María G., Woelders, Henri, and Blesbois, Elisabeth

Abstract

Seminal plasma is a key biological fluid that modulates sperm function in the reproduction process. However, its role in sperm biotechnologies is scarce in poultry. The aims of the present study were to study the amino acids profile and total proteins of seminal plasma in 12 Spanish chicken breeds and to investigate the role of seminal plasma on cryoresistance of rooster sperm. To investigate the role of seminal plasma on cryoresistance, diluted pooled semen samples were cryopreserved in the presence and absence of seminal plasma. Glutamic acid was the most abundant free amino acid in seminal plasma, followed by alanine, serine, valine, and glycine. There was an influence of breed (P<0.05) on the percentage of viable sperm after freezing-thawing of samples with seminal plasma. Cluster analysis revealed that White Prat, Black Castellana, Blue Andaluza, Quail Castellana, and Red-Barred Vasca returned the best freezing-thawing response (good freezers). There was a positive correlation between seminal plasma concentrations of valine, isoleucine lysine, leucine and post thaw viability. The evaluation of fertilization capacity of frozen-thawed semen from the breeds White Prat (‘good freezer’) and Black-Red Andaluza (‘bad freezer’) showed that good freezer had higher fertility (20/68, 29.4%) compared to bad freezer breed (14/76, 18.4%), even if the difference was not significant (P = 0.08). The TUNEL assay revealed that freezing/thawing procedures in presence of seminal plasma provoked higher DNA fragmentation in most of the breeds, with a positive correlation between seminal alanine, valine, isoleucine, methionine, leucine, tyrosine, phenylalanine concentrations and DNA integrity. DNA fragmentation was lower in absence of seminal plasma and the breed effect on sperm viability was highly reduced. It is concluded that specific seminal plasma amino acids were associated with post-thaw percentage of viable sperm and DNA integrity. The removal of seminal plasma decreases th

Published
2019

41. Access to pasture in an outdoor housing system affects welfare indicators and improves rooster sperm quality in two native Mediterranean breeds

Author

Santiago-Moreno, Julian and Blesbois, Elisabeth

Subjects
semen motility, urogenital system, chicken, outdoor system
Abstract

A paper produced in the frame of the H2020 project 677353 'Innovative Management of Animal GEnetic resources'. The study explores the effect of rooster management on sperm quality, and shows a favorable effect of an outdoor system on the motilityoffresh semen, but not on the motility offrozen/thawed semen.

Published
2018

42. Chicken semen cryopreservation and use for the restoration of rare genetic resources

Author

Thélie, Aurore, Bailliard, Amélie, Seigneurin, François, Zerjal, Tatiana, Tixier-Boichard, Michèle, Blesbois, Elisabeth, Physiologie de la reproduction et des comportements [Nouzilly] (PRC), Institut National de la Recherche Agronomique (INRA)-Institut Français du Cheval et de l'Equitation [Saumur]-Université de Tours-Centre National de la Recherche Scientifique (CNRS), Syndicat des Sélectionneurs Avicoles et Aquacoles Français (SYSAAF), Génétique Animale et Biologie Intégrative (GABI), Institut National de la Recherche Agronomique (INRA)-AgroParisTech, PIA CRB-Anim, and Institut National de la Recherche Agronomique (INRA)-Institut Français du Cheval et de l'Equitation [Saumur]-Université de Tours (UT)-Centre National de la Recherche Scientifique (CNRS)

Subjects
Male, endocrine system, preservation, cryobanks, [SDV.BIO]Life Sciences [q-bio]/Biotechnology, poulet, Biotechnologies, glycerol, Cryoprotective Agents, Freezing, Animals, Insemination, Artificial, ressource génétique animale, Cryopreservation, capacité de reprodution, cryoconservateur, urogenital system, cryoconservation du sperme, Sperm Banks, fertilité animale, Genetic Variation, stockage à long terme, Spermatozoa, Fertility, stockage de semence, Female, Chickens, Semen Preservation
Abstract

International audience; For the past 50 yr, practices for ex situ preservation of endangered breeds have been extended. Semen and primordial germ cells, gonadic tissues have been frozen to create genetic stocks of chicken genetic diversity in cryobanks. Semen cryopreservation stays the preferred method since it is not invasive. Many protocols have been developed to cryopreserve chicken semen, but they give highly variable success rate. The aim of the present study was to standardize and prove the effectiveness of semen long-term storage for the restitution of lost families. We showed that semen straws stored for 18 yr in liquid nitrogen did not lose their fertilizing ability. We demonstrated the usefulness of cryopreserved semen stored in the French National Cryobank for the recovery of families of a subfertile experimental chicken line. In order to highlight the standardization of the cryopreserved method, different cryoprotectant protocols were also tested on a rare breed, freezing/thawing and insemination conditions were controlled. The best results were obtained using glycerol protocol, a sperm dilution of 1:4 (semen:extender). The insemination dose of 200 million sperm/female was as efficient as 400 million of sperm. Altogether, these results demonstrated the effectiveness of chicken semen long-term storage for the restoration of lost genetic resources and highlighted the importance of standardized chicken semen cryopreservation using procedures combining biophysical (cryoprotectants, freezing/thawing conditions) and zootechnical (artificial insemination) features.

Published
2018

46. Seminal plasma amino acid profile in different breeds of chicken: Role of seminal plasma on sperm cryoresistance

Author

Santiago-Moreno, Julián, primary, Bernal, Berenice, additional, Pérez-Cerezales, Serafín, additional, Castaño, Cristina, additional, Toledano-Díaz, Adolfo, additional, Esteso, Milagros C., additional, Gutiérrez-Adán, Alfonso, additional, López-Sebastián, Antonio, additional, Gil, María G., additional, Woelders, Henri, additional, and Blesbois, Elisabeth, additional

Published
2019
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50. Search for the genes involved in oocyte maturation and early embryo development in the hen

Author

Blesbois Elisabeth, Monget Philippe, Martin-Magniette Marie-Laure, Balzergue Sandrine, Couty Isabelle, Batellier Florence, Elis Sebastien, and Govoroun Marina S

Subjects
Biotechnology, TP248.13-248.65, Genetics, QH426-470
Abstract

Abstract Background The initial stages of development depend on mRNA and proteins accumulated in the oocyte, and during these stages, certain genes are essential for fertilization, first cleavage and embryonic genome activation. The aim of this study was first to search for avian oocyte-specific genes using an in silico and a microarray approaches, then to investigate the temporal and spatial dynamics of the expression of some of these genes during follicular maturation and early embryogenesis. Results The in silico approach allowed us to identify 18 chicken homologs of mouse potential oocyte genes found by digital differential display. Using the chicken Affymetrix microarray, we identified 461 genes overexpressed in granulosa cells (GCs) and 250 genes overexpressed in the germinal disc (GD) of the hen oocyte. Six genes were identified using both in silico and microarray approaches. Based on GO annotations, GC and GD genes were differentially involved in biological processes, reflecting different physiological destinations of these two cell layers. Finally we studied the spatial and temporal dynamics of the expression of 21 chicken genes. According to their expression patterns all these genes are involved in different stages of final follicular maturation and/or early embryogenesis in the chicken. Among them, 8 genes (btg4, chkmos, wee, zpA, dazL, cvh, zar1 and ktfn) were preferentially expressed in the maturing occyte and cvh, zar1 and ktfn were also highly expressed in the early embryo. Conclusion We showed that in silico and Affymetrix microarray approaches were relevant and complementary in order to find new avian genes potentially involved in oocyte maturation and/or early embryo development, and allowed the discovery of new potential chicken mature oocyte and chicken granulosa cell markers for future studies. Moreover, detailed study of the expression of some of these genes revealed promising candidates for maternal effect genes in the chicken. Finally, the finding concerning the different state of rRNA compared to that of mRNA during the postovulatory period shed light on some mechanisms through which oocyte to embryo transition occurs in the hen.

Published
2008
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