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Progesterone (P4) is predicted to act as a negative regulatory hormone for oocyte maturation events; however, its local effects during follicular development remain poorly understood in bovine. The complex process of oocyte meiosis progression is dependent on cellular communication among follicular cells. Besides, the breakdown of this communication, mainly between cumulus cells (CC) and oocyte, through the retraction of cumulus projections connecting these cells can impact oocyte maturation. In our study, we observed that follicles from the ovary ipsilateral to the corpus luteum (CL) containing high intrafollicular P4 concentrations enhance the abundance of proteins detected in follicular-derived small extracellular vesicles (sEVs) predicted to be involved in the retraction of membrane projections based on actin filaments, such as transzonal projections (TZPs). Conversely, we found that follicles from the ovary contralateral to the CL, which contained low intrafollicular P4 concentrations, had a high detection of proteins predicted to regulate the maintenance of TZPs. We also performed RNAseq analysis which demonstrated that 177 genes were differentially expressed in CC under the different P4 environments. Bioinformatic analysis points to changes associated to cell metabolism in cells from follicles ipsilateral to the CL in comparison to genes involved in cell communication in CC from follicles contralateral to the CL. Our functional analysis experiment confirmed that supplementation of cumulus-oocyte complexes during in vitro maturation with P4 at concentration similar to ipsilateral follicles reduces the number of TZPs. In summary, our study underscores a direct association between P4 concentration and cumulus-oocyte interaction, with potential consequences for the acquisition of oocyte competence., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Juliano Coelho da Silveira reports financial support was provided by São Paulo Research Foundation - FAPESP. Juliano Coelho da Silveira reports financial support was provided by Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - Brasil (CAPES). Juliano Coelho da Silveira reports financial support was provided by National Council for Scientific and Technological Development. If there are other authors, they declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

The risk of chronic diseases such as cardiovascular disease, cancer, inflammation, obesity, and autoimmune disease is linked to the quality of dietary fats, with lower intake of saturated and higher intake of n-6 and n-3 polyunsaturated fatty acids (PUFA) considered beneficial to health. This study investigated the effect of supplementing sows' diets with cow's milk biofortified with n-6 or n-3 PUFA, at varying n-6/n-3 ratios (8.26, 7.92, and 2.72) during their growing phase and throughout gestation and lactation on their reproductive performance and immune-inflammatory status. Specifically, we analyzed circulating cholesterol and fatty acid profiles of serum, colostrum and milk, sow body weight, and neonate colostrum intake, Apgar scores, muscle composition, and embryo viability. Analysis of circulating immunoglobulins (Ig), interleukins, and eicosanoids and complement system hemolytic activity were used to evaluate inflammatory and immune responses of sows and piglets. Expression of lipolysis and lipogenic genes in the liver were investigated in sows and piglets, with additional investigation of hypothalamus genes regulating appetite in sows. Feeding sows milk biofortified with n-6 and n-3 PUFA altered serum fatty acid profiles, reduced triglycerides (TG), increased embryo total number, increased early gestation backfat, and reduced colostrum IgG. Piglets of biofortified sow had higher circulating IgA, IgM and TNF-α, and lower IL-10. Sows fed n-3 biofortified milk had higher very low-density lipoproteins (VLDL) and TNF-α in circulation. Offspring from sows fed n-6 versus n-3 biofortified milk had lower IL-10 and expression levels of SREBP-1. N-3 versus n-6 also lowered arachidonic acid (ARA) levels in sow's milk and piglet viability 1. Findings offer insights into the potential health benefits of dietary supplementation with biofortified milk in swine, which serve as good model of diet-nutrition studies of humans, and therefore can potentially be considered in dietary recommendations both human and animal populations., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Reis et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

In cattle, 17β-estradiol (E2) is essential for triggering luteolysis via the synthesis of prostaglandin F2α (PGF2α). We aimed to evaluate the effects of E2-treatment on day 15 of the estrous cycle on the transcript abundance of genes involved in the PGF2α synthetic cascade. Nelore heifers (N = 50) were subjected to a hormonal protocol for the synchronization of ovulation. Between days 14 and 23 after estrus, the area (cm 2 ) and blood perfusion (%) of the corpus luteum (CL) and progesterone (P4) plasma concentrations were evaluated daily. On day 15, the heifers were assigned to the Control (2 mL of pure sesame oil, N = 21) or Estradiol group (1 mg of E2 diluted in 2 mL of sesame oil, N = 23). After the treatments at 0 h, uterine biopsies were collected at times 1.5 h (C1.5h, N = 8 and E1.5h, N = 10) or 3 h (C3h, N = 8 and E3h, N = 11); and blood samples were obtained from 0, 3, 4, 6 and 7 h for the measurement of 13,14-dihydro-15-keto-PGF2α (PGFM) concentrations by ELISA. Transcript abundance was determined by RT-qPCR and protein abundance of ESRβ and OXTR was determined by Western Blotting. The Estradiol group showed greater (P < 0.05) concentrations of PGFM at 6 and 7 h compared to the Control group. A progressive decrease in plasma P4 concentrations characterized a hastened functional luteolysis, followed by structural luteolysis in the Estradiol group (P < 0.05). Among the treatment groups, no significant difference was detected for the abundance of PRKCα, PRKCβ, AKR1B1, PTGS2 and ESRα transcripts (P > 0.05). Estradiol treatment decreased the abundance of PLA2G4A, AKR1C4, and ESRβ both 1.5h and 3h after treatment (P < 0.05). The relative expression of PGR and OXTR was greater in E3h compared to the C3h (P > 0.05). Protein abundance did not differ between treatment groups at either experimental times (P > 0.05). Overall, E2 promoted an increase in PGFM concentrations and the hastening of functional and structural luteolysis in Nelore heifers through the upregulation of PGR and OXTR, demonstrating for the first time that the expression of these receptors within 3 h after E2 stimulus was associated with triggering luteolysis in cattle., (Copyright © 2023 Elsevier Inc. All rights reserved.)

The stem cell-based research for reproductive biotechnology has been widely studied and shows promise for repairing defective tissue or degenerated cells to treat different diseases. The adipose tissue and amniotic membrane have awakened great interest in regenerative medicine and arises as a promising source of mesenchymal stem cells. Both types, adipose and amniotic derived mesenchymal stem cells (AMSCs) are multipotent cells with an enhanced ability to differentiate into multiple lineages.. We aimed to evaluate the effect of basal supplementation of exosomes in cell cultures with canine amniotic mesenchymal stem cells (MSCs). Mesenchymal stem cells derived from canine amniotic and adipose tissue were isolated and cultured performing cell passages until 80-90% confluence was reached. The growth curve was determined and peak cell growth was observed in the second passage. The cells were then characterized and differentiated into adipogenic, chondrogenic and osteogenic lineages. Extracellular vesicles from amnion were isolated using an ultracentrifugation protocol and characterized by nanosight analysis. To evaluate their ability to improve cellular viability in naturally inefficient passages, exosomes were co-cultures to the MSC cells. The results showed a 15-20% increase in the expansion rate of cultures supplemented with vesicles extracted in the first and second passages when compared to the control group. Statistical analysis using the Dunnett test (p ≤ 0.05) corroborated this result, showing a positive correlation between supplementation and expansion rate. These results indicate not only the importance of exosomes in the cell communication process but also the feasibility of the culture supplementation protocol for therapeutic purposes. The potential of the AMSCs for reproductive biotechnology is undoubted, however, their application to repair reproductive disorders and the involved mechanisms remain elusive. The strategies to enable the Adipose Stem Cells and AMSCs application in reproductive biotechnology and optimize their use for tissue regeneration open new venues using exosomes interactions., Competing Interests: Declaration of competing interest The authors declare that there are no conflicts of interest on this reports research., (Copyright © 2022 Elsevier Inc. All rights reserved.)

The oviduct is an important reproductive structure that connects the ovary to the uterus and takes place to important events such as oocyte final maturation, fertilization and early embryonic development. Thus, gametes and embryo can be directly influenced by the oviductal microenvironment composed by epithelial cells such secretory and ciliated cells and oviductal fluid. The oviduct composition is anatomically dynamic and is under ovarian hormones control. The oviductal fluid provides protection, nourishment and transport to gametes and embryo and allows interaction to oviductal epithelial cells. All these functions together allows the oviduct to provides the ideal environment to the early reproductive events. Extracellular vesicles (EVs) are biological nanoparticles that mediates cell communication and are present at oviductal fluid and plays an important role in gametes/embryo - oviductal cells communication. This review will present the ability of the oviducts based on its dynamic and systemic changes during reproductive events, as well as the contribution of EVs in this process., Competing Interests: Conflicts of interest: The authors have no conflict of interest to declare.

Freeze boar semen is still the biggest challenge for the swine industry due to the high cold shock sensitivity of boar sperm cells and the variance of post-thaw results among individuals and ejaculates from the same boar. To solve this problem, we investigate if miRNAs present in sperm cells and small extracellular vesicles (EVs) from seminal plasma of raw boar ejaculates can predict high-quality ejaculates after underwent the freeze-thaw process. For this, we obtained miRNAs samples of sperm cells and EVs from raw seminal plasma from 27 ejaculates before the cryopreservation process. Two groups with different freezability considering the analysis post-thaw of structure and sperm functionality were formed: High freezability (HF; n = 04) and low freezability (LF; n = 04). That done, we investigated the miRNAs profile of sperm cells and EVs from seminal plasma in both groups. Three miRNAs were differently abundant in LF ejaculates, being the ssc-miR-503 found in higher levels in sperm cells (P < 0.10). The ssc-miR-130a and ssc-miR-9 most abundant in EVs from seminal plasma (P < 0.10), in LF ejaculates. Through enrichment analysis, it was possible to verify that these miRNAs could be performing modifications in the development of male germ cells and in the production of energy to spermatozoa to maintain their viability and functionality. Therefore, we can demonstrate that ssc-miR-503, ssc-miR-130a, and ssc-miR-9 are related to low sperm cryotolerance in boars semen. So those miRNAs can be used as a biomarker to predict their low ability to tolerate the cryopreservation process., Competing Interests: Declaration of competing interest The authors declare that there is no conflict of interest that could be perceived as prejudicing the impartiality of the research reported., (Copyright © 2021 Elsevier Inc. All rights reserved.)

Information on molecular mechanisms through which sex-steroids regulate oviductal function to support early embryo development is lacking. Here, we hypothesized that the periovulatory endocrine milieu affects the miRNA processing machinery and miRNA expression in bovine oviductal tissues. Growth of the preovulatory follicle was controlled to obtain cows that ovulated a small follicle (SF) and subsequently bore a small corpus luteum (CL; SF-SCL) or a large follicle (LF) and large CL (LF-LCL). These groups differed in the periovulatory plasmatic sex-steroid's concentrations. Ampulla and isthmus samples were collected on day four of the estrous cycle. Abundance of DROSHA, DICER1, and AGO4 transcripts was greater in the ampulla than the isthmus. In the ampulla, transcription of these genes was greater for the SF-SCL group, while the opposite was observed in the isthmus. The expression of the 88 most abundant miRNAs and 14 miRNAs in the ampulla and 34 miRNAs in isthmus were differentially expressed between LF-LCL and SF-SCL groups. Integration of transcriptomic and miRNA data and molecular pathways enrichment showed that important pathways were inhibited in the SF-SCL group due to miRNA control. In conclusion, the endocrine milieu affects the miRNA expression in the bovine oviduct in a region-specific manner.