Abstract

Understanding the composition of the oviduct fluid (OF) is crucial to better comprehend the microenvironment in which sperm capacitation, fertilization and early embryo development take place. Therefore, our aim was to determine the spatiotemporal changes in the OF proteome according to the anatomical region of the oviduct (ampulla vs. isthmus), the proximity of the ovulating ovary (ipsilateral vs. contralateral side) and the peri-ovulatory stage (pre-ovulatory or Pre-ov vs. post-ovulatory or Post-ov). Oviducts from adult cyclic cows were collected at a local slaughterhouse and pools of OF were analyzed by nanoLC-MS/MS and label-free protein quantification (n = 32 OF pools for all region × stage × side conditions). A total of 3760 proteins were identified in the OF, of which 65% were predicted to be potentially secreted. The oviduct region was the major source of variation in protein abundance, followed by the proximity of the ovulating ovary and finally the peri-ovulatory stage. Differentially abundant proteins between regions, stages and sides were involved in a broad variety of biological functions, including protein binding, response to stress, cell-to-cell adhesion, calcium homeostasis and the immune system. This work highlights the dynamic regulation of oviduct secretions and provides new protein candidates for interactions between the maternal environment, the gametes and the early embryo.

Highlights

  • The objective of this study was to determine the effect on the oviductal fluid (OF) proteome of (1) the region, (2) the proximity of the ovulating ovary and (3) the peri-ovulatory stage (Pre-ov vs. Post-ov) using nanoLC-MS/MS analysis

  • A total of 3760 proteins were identified in the bovine OF (Supplementary Data S1)

  • The present study provides an extensive analysis of the bovine OF proteome considering three factors of variation, i.e., the anatomical oviduct region, the peri-ovulatory stage and the proximity of the ovulating ovary

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Summary

Introduction

Proteins were reported to vary in abundance between cyclic and pregnant cows on day 3 post-estrus, with slight differences between the ampulla and i­sthmus[14]. Region-related differences in oviductal secretions were reported in vitro: effects of OF-derived extracellular vesicles (EVs)[15] and OF p­ roteins[16] on sperm viability and capacitation differed between ampulla and isthmus. The quality of embryos developed in vitro with bovine OF-derived ­EVs17 or ovine ­OECs18 differed depending on the ampullary or isthmic origin of the EVs or OECs. The spatiotemporal variation in OF proteome remains unclear. A dynamic variation between the ampulla and isthmus in the oviduct ipsilateral to ovulation, and between the Pre-ov and Post-ov stages are expected, in response to the specific requirements of gametes and embryos. The objective of this study was to determine the effect on the OF proteome of (1) the region (isthmus vs ampulla), (2) the proximity of the ovulating ovary (ipsilateral vs contralateral side) and (3) the peri-ovulatory stage (Pre-ov vs Post-ov) using nanoLC-MS/MS analysis

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