Spatiotemporal profiling of the bovine oviduct fluid proteome around the time of ovulation

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.

The sperm-interacting proteome in bovine oviduct fluid: temporal and spatial resolution

“…Of the 245 SIPs, 78% were previously reported in oviductal EVs, including exosomes and microvesicles. This proportion was much higher than those reported for proteins identi ed in the whole bovine OF [14], indicating an enrichment in EV-secreted proteins among SIPs. Interactions between spermatozoa and oviduct EVs have already been described in cats [34], cattle [10], pigs [44], and mice [45].…”

“…All proteins containing at least two unique peptides with an FDR < 0.01% were considered for protein quanti cation. Protein abundance was assessed using the Spectrum Count quantitative method and the "weighted spectra" option of the Scaffold software, as previously described [14]. The weighted spectra option of Scaffold is based on the assignment of each peptide to a weight according to whether they are shared or not between identi ed proteins.…”

“…The bovine OF proteome is highly regulated according to the estrous cycle stage [9,13]. In addition, recent work from our laboratory showed that the oviduct anatomical region (isthmus vs. ampulla) also has a signi cant impact on the OF proteome [14].…”

The proteomic analysis of bovine embryos developed in vivo or in vitro reveals the contribution of the maternal environment to early embryo

“…Some other proteins found overabundant in the in vivo -derived embryos were previously identified as interacting with in vitro -produced embryos after incubation with post-ovulatory oviduct fluid, including retinal dehydrogenase 1 (ALDH1A1) and alpha isoform of regulatory subunit A protein phosphatase 2 (PPP2R1A) at the 4–6 cell stage, cytoplasmic aconitate hydratase (ACO1), hemoglobin subunit beta (HBB) and purine nucleoside phosphorylase (PNP) at the morula stage, and annexin A8 (ANXA8), creatine kinase B-type (CKB), chloride intracellular channel protein 1 (CLIC1), cytosolic non-specific dipeptidase (CNDP2), hemoglobin subunit alpha (HBA), ribonuclease inhibitor (RNH1), tubulin polymerization-promoting protein family member 3 (TPPP3), and CD109 at both stages [ 25 ]. A comparison between proteins in cluster 1 and those identified by MS in the isthmic part of the oviduct fluid just after ovulation [ 59 ], i.e., place and time of early embryo development, indicates that 341 (75%) are also present in the oviduct fluid (data not shown). Among those, OVGP1, CD109, CLU, RARRES1, and tubulin beta-4B chain (TUBB4B) were quantified at high abundance in the postovulatory oviduct fluid (36 to 121 NWS, [ 59 ]) and were found overabundant in in vivo -derived embryos from the 4–6 cell up to blastocyst stages, making them good candidates for embryo-interacting proteins originated in the oviduct.…”

An interactive analysis of the mouse oviductal miRNA profiles

“…Developmental control is then passed from the mother to the zygote ( Hamm and Harrison, 2018 ). In addition, a recent comprehensive characterization of the bovine OF proteome revealed a spatiotemporal regulation of the OF according to the anatomical region of the oviduct, the proximity to the ovulating ovary and the stage of the cycle ( Mahé et al, 2022 ). Future approaches should investigate the potential involvement of miRNA in the fine-tune regulation of the oviductal microenvironment.…”