Estrous cycle impacts microRNA content in extracellular vesicles that modulate bovine cumulus cell transcripts during in vitro maturation†

Abstract

Extracellular vesicles (EVs) are nanoparticles secreted by ovarian follicle cells. Extracellular vesicles are an important form of intercellular communication, since they carry bioactive contents, such as microRNAs (miRNAs), mRNAs, and proteins. MicroRNAs are small noncoding RNA capable of modulating mRNA translation. Thus, EVs can play a role in follicle and oocyte development. However, it is not clear if EV contents vary with the estrous cycle stage. The aim of this study was to investigate the bovine miRNA content in EVs obtained from follicles at different estrous cycle stages, which are associated with different progesterone (P4) levels in the follicular fluid (FF). We collected FF from 3 to 6 mm follicles and evaluated the miRNA profile of the EVs and their effects on cumulus-oocyte complexes during in vitro maturation. We observed that EVs from low P4 group have a higher abundance of miRNAs predicted to modulate pathways, such as MAPK, RNA transport, Hippo, Cell cycle, FoxO, oocyte meiosis, and TGF-beta. Additionally, EVs were taken up by cumulus cells and, thus, affected the RNA global profile 9 h after EV supplementation. Cumulus cells supplemented with EVs from low P4 presented upregulated genes that could modulate biological processes, such as oocyte development, immune responses, and Notch signaling compared with genes of cumulus cells in the EV free media or with EVs from high P4 follicles. In conclusion, our results demonstrate that EV miRNA contents are distinct in follicles exposed to different estrous cycle stage. Supplementation with EVs impacts gene expression and biological processes in cumulus cells.

Post-Translational Protein Deimination Signatures in Serum and Serum-Extracellular Vesicles of Bos taurus Reveal Immune, Anti-Pathogenic, Anti-Viral, Metabolic and Cancer-Related Pathways for Deimination

“…F95-enrichment for deiminated proteins from serum and serum-EVs revealed a range of immunological, metabolic and gene regulatory proteins as candidates for this post-translational modification, therefore indicating hitherto under-recognized modes for protein-moonlighting of these proteins in bovine immunity and physiology. Few studies have hitherto assessed roles for PADs and deimination in cattle [22,23,42], while a range of studies have been carried out on EVs in relation to cattle immunity, fertility and development [54,57,60,61,63,65,68,74,78,83,85,88,90]. Export of post-translationally modified proteins, such as deiminated proteins in the current study, has not been assessed before in cattle serum.…”

Liquid biopsy based on small extracellular vesicles predicts chemotherapy response of canine multicentric lymphomas

“…Particle size and concentration was determined using nanoparticle tracking analysis (NTA). The protocols were based on previous work and briefly described above 59 , 60 .…”

“…The transmission electron microscopy was performed according Avila and colleagues 60 . Briefly, exosome pellets isolated from 200 µl of serum of dogs from control and lymphoma group were diluted in 50 μl of fixation solution (0.1 M cacodylate; 2.5% glutaraldehyde and 4% paraformaldehyde at pH 7.2–7.4) for 2 h at 4℃.…”

Plasma extracellular vesicle miRNAs as potential biomarkers of superstimulatory response in cattle

“…Both miRNAs play a role in regulating luteal steroidogenesis in bovine luteal tissues 72 and human follicular cells 73 . Recently, de Ávila et al 74 investigated the EV-miRNA content of bovine follicles at different stages of the estrous cycle in association with different follicular fluid P4 concentrations. A group of EV-miRNAs was up-regulated in the follicular fluids of the low compared with the high P4 group.…”