Spermatozoa induce transcriptomic alterations in bovine oviductal epithelial cells prior to initial contact

Abstract

The capability of spermatozoa to directly influence maternal gene expression is already established. Indeed, some of the changes induced by spermatozoa may have a direct functional importance in the pre-conceptional period. Although the mechanisms underlying these sperm-maternal interactions are not well characterized, it is possible that they could involve ligands that are released from the spermatozoa. This study therefore aimed to test whether physical contact between bovine spermatozoa and bovine oviductal epithelial cells (BOECs) is a prerequisite for spermatozoa-induced gene expression changes. We used two co-culture models: a contact co-culture model in which spermatozoa interact directly with BOECs, and a non-contact co-culture model in which an insert with the pore size of 0.4 μm was placed between spermatozoa and BOECs. Messenger RNA sequencing analysis of BOECs by RNA-seq revealed ten differentially expressed genes in contact system and 108 differentially expressed genes in the non-contact system after 10 h of co-culture. Retinol metabolism pathway and ovarian steroidogenesis pathway were significantly enriched in the non-contact co-culture system. Q-PCR analysis revealed that transcriptional responses can be rapid, with increased expression of four genes (DHRS3, CYP1B1, PTGS2, and ATF3) detectable within just 90 min of co-incubation, but with expression levels highly dependent on the type of co-culture system. The findings from our study demonstrate that direct contact with spermatozoa is not necessary to induce changes in gene expression of oviductal epithelial cells, suggesting that spermatozoa may be able to signal to maternal tissues in advance of their arrival.

Bovine Follicular Fluid Derived Extracellular Vesicles Modulate the Viability, Capacitation and Acrosome Reaction of Bull Spermatozoa

“…Frozen semen straws from three different bulls were processed separately following the methods described by our group previously [ 36 ]. In brief, frozen semen straws (4 straws from each bull) were thawed at 37 °C and the contents of the straws were layered over 4 mL of 60% isoosmotic Percoll ® solution (GE Healthcare, 17-0891-02, Vendevägen, Sweden) Subsequently, the overlayered semen samples were centrifuged at 300× g for 20 min at RT.…”

Extracellular vesicles in seminal plasma: A safe and relevant tool to improve fertility in livestock?

“…It appears logical to state that the SP is a composite secretion that promotes cell-to-cell communication between reproductive and non-reproductive cells, following evidence from experimental studies in vivo (Alvarez-Rodriguez et al, 2019a;2020a) and in vitro (Reshi et al, 2020). Of interest, however, is to insist in the fact that sEVs are, at least in vitro (Bai et al, 2018), able to elicit specific changes in the lining epithelium.…”

Role of extracellular vesicles in intercellular communication during reproduction

“…Studies showed that EVs produced from the male reproductive tract, including epididymosomes and prostasomes, play vital roles in the maturation process of spermatozoa (Saez et al, 2003; Wu et al, 2017). Spermatozoa undergo several physical and biochemical changes during their journey towards oocyte starting from the anterior vagina after coitus (Reshi et al, 2020). These modifications possibly occur due to the crosstalk between the spermatozoa and the female reproductive tract leading to functional maturation.…”