Protein Cargo of Extracellular Vesicles From Bovine Follicular Fluid and Analysis of Their Origin From Different Ovarian Cells

Uzbekova, S.; Almiñana, C.; Labas, V.; Teixeira-Gomes, A.; Combes-Soia, L.; Tsikis, G.; Carvalho, A.V.; Uzbekov, R.; Singina, G.

Авторы: Uzbekova Svetlana, Almiñana Carmen, Labas Valerie, Teixeira-Gomes Ana-Paula, Combes-Soia Lucie, Tsikis Guillaume, Carvalho Anais Vitorino, Uzbekov Rustem, Singina Galina
Журнал: Frontiers in Veterinary Science
Том: 7
Год издания: 2020
Издательство: Frontiers Media S.A.
Местоположение издательства: Lausanne, Switzerland
Номер статьи: 584948
DOI: 10.3389/fvets.2020.584948
Аннотация: fluid (FF) fills the interior portion of the ovarian antral follicle and provides asuitable microenvironment for the growth of the enclosed oocyte through molecularfactors that originate from plasma and the secretions of follicular cells. FF containsextracellular nanovesicles (ffEVs), including 30–100-nm membrane-coated exosomes,which carry different types of RNA, proteins, and lipids and directly influence oocytecompetence to develop embryo. In the present study, we aimed to characterize theprotein cargo of EVs from the FF of 3–6-mm follicles and uncover the origins of ffEVs byassessing expression levels of corresponding mRNAs in bovine follicular cells and oocyteand cell proteomes. Isolated exosome-like ffEVs were 53.6 + 23.3 nm in size and couldbe internalized by cumulus-oocyte complex. Proteomes of ffEVs and granulosa cells(GC) were assessed using nanoflow liquid chromatography coupled with high-resolutiontandemmass spectrometry after the gel fractionation of total proteins. In total, 460 proteinisoforms corresponding to 322 unique proteins were identified in ffEVs; among them,190 were also identified via GC. Gene Ontology terms related to the ribosome, proteinand RNA folding, molecular transport, endocytosis, signal transduction, complementand coagulation cascades, apoptosis, and developmental biology pathways, includingPI3K-Akt signaling, were significantly enriched features of ffEV proteins. FfEVs containnumerous ribosome and RNA-binding proteins, which may serve to compact differentRNAs to regulate gene expression and RNA degradation, and might transfer ribosomalconstituents to the oocyte. Majority of genes encoding ffEV proteins expressed atdifferent levels in follicular cells and oocyte, corroborating with numerous proteins, whichwere reported in bovine oocyte and cumulus cells in other studies thus indicating possibleorigin of ffEV proteins. The limited abundance of several mRNAs within follicular cellsindicated that corresponding ffEV proteins likely originated from circulating exosomesreleased by other tissues. Analysis of bovine ffEV transcriptome revealed that mRNAs present in ffEV accounted for only 18.3% of detected ffEV proteins. In conclusion, ourstudy revealed numerous proteins within ffEVs, which originated from follicular and othercells. These proteins are likely involved in the maintenance of follicular homeostasis andmay affect oocyte competence.
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Protein Cargo of Extracellular Vesicles From Bovine Follicular Fluid and Analysis of Their Origin From Different Ovarian Cellsстатья

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