Dataset Information

The proteome of equine oviductal fluid varies before and after ovulation: a comparative study

ABSTRACT: Equine fertilization cannot be performed in the laboratory as equine spermatozoa do not cross the oocyte´s zona pellucida in vitro. Hence, a more profound study of equine oviductal fluid (OF) composition at the pre-ovulatory and post-ovulatory stages could help to understand what components are required to achieve fertilization in horses. Our work aimed to elucidate the proteomic composition of equine OF at both stages. To do this, OF was obtained postmortem from oviducts of slaughtered mares ipsilateral to a pre-ovulatory follicle (n = 4) or a recent ovulation (n = 4); the samples were kept at -80 °C until analysis. After protein extraction and isobaric tags for relative and absolute quantification (iTRAQ) labelling, the samples were analyzed by nanoliquid chromatography coupled to tandem mass spectrometry (LC-MS/MS). The analysis of the spectra resulted in the identification of a total of 1173 proteins present in pre-ovulatory and post-ovulatory samples; among these, 691 were unique for Equus caballus. Proteins from post-ovulatory oviductal fluid were compared with proteins from pre-ovulatory oviductal fluid and were categorized as up- (positive log fold change) or down-regulated (negative log fold change); 15 proteins were found to be down-regulated in the post-ovulatory fluid and 154 were up-regulated in the post-ovulatory OF compared to the pre-ovulatory fluid. The identified proteins were related to sperm-oviduct interaction, fertilization, and metabolism, among others. Our data reveal consistent differences in the proteome of equine OF prior and after ovulation, helping to increase our understanding in the factors that promote fertilization and early embryo development in horses.

INSTRUMENT(S): TripleTOF 5600

ORGANISM(S): Equus Caballus (horse)

TISSUE(S): Oviductal Fluid

SUBMITTER: Antonio Ramos Fernández

LAB HEAD: Beatriz Macías García

PROVIDER: PXD025320 | Pride | 2022-02-17

REPOSITORIES: Pride

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Publications

The Proteome of Equine Oviductal Fluid Varies Before and After Ovulation: A Comparative Study.

Fernández-Hernández Pablo P Marinaro Federica F Sánchez-Calabuig María Jesús MJ García-Marín Luis Jesús LJ Bragado María Julia MJ González-Fernández Lauro L Macías-García Beatriz B

Frontiers in veterinary science 20210805

Equine fertilization cannot be performed in the laboratory as equine spermatozoa do not cross the oocyte's zona pellucida <i>in vitro</i>. Hence, a more profound study of equine oviductal fluid (OF) composition at the pre-ovulatory and post-ovulatory stages could help in understanding what components are required to achieve fertilization in horses. Our work aimed to elucidate the proteomic composition of equine OF at both stages. To do this, OF was obtained postmortem from oviducts of slaughtere ...[more]

PMID: 34422946

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Project description:The success of the early reproductive events depends on an appropriate communication between gametes/embryos and the oviduct. Extracellular vesicles (EVs) contained in oviductal secretions have been suggested as new players in mediating this crucial cross-talk by transferring their cargo (e.g. proteins, mRNA and small ncRNA) from cell to cell. However, little is known about the oviduct EVs composition and their implications in reproductive success. The objective of our study was to determine the changes of oviductal EVs small ncRNA content under the hormonal influence during the estrous cycle. Methods: EVs, exosomes and microvesicles, were isolated from bovine oviductal fluid at different stages of the estrous cycle (postovulatory-stage, early luteal phase, late luteal phase and pre-ovulatory stage). RNA enriched for short molecules was isolated and used for the preparation of small RNA-seq libraries. RNA-sequencing was performed on an Illumina HiSeq 2500. The obtained sequence reads that passed quality filters were annotated by BLAST alignment to various sets of non-coding and coding RNAs. Read counts were calculated based on the frequency of unique sequences after removal of PCR duplicates. Statistical analysis was done with EdgeR to identify differential small ncRNA abundance across the different stages of the estrous cycle. Results: Small RNA-Seq identified the presence of different types of ncRNAs (miRNAs, rRNA fragments, tRNA fragments, snRNA, snoRNA, and other ncRNAs), which were partially also under hormonal influence. Major differences were found between post-ovulatory and early luteal and the rest of the stages for small ncRNAs. Interesting miRNAs identified in oviductal EVs, miR-34b and miR-449a, showed differential abundance among stages. Dysregulation of these miRNAs has been associated with defective cilia in the oviduct and infertility. Conclusions: Our findings represent the first extensive oviductal signature of bovine oviductal EVs miRNA content and contribute to a better understanding of the role of EVs as modulators of gamete/embryo-maternal interactions.

Dataset Information

The proteome of equine oviductal fluid varies before and after ovulation: a comparative study

Publications

The Proteome of Equine Oviductal Fluid Varies Before and After Ovulation: A Comparative Study.

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