Relative abundance of extra-cellular miRNAs in bovine follicular fluid: Implication for cell–cell communication during oocyte growth

Abstract

Here we aimed to investigate the expression patternof the circulating extra-cellular miRNAs in exosome andnon-exosomal fraction of follicular fluid consisted of fullygrown or growing oocytes and to validate exosome medi-ated Here we aimed to investigate the expression patternof the circulating extra-cellular miRNAs in exosome andnon-exosomal fraction of follicular fluid consisted of fullygrHere we aimed to investigate the expression patternof the circulating extra-cellular miRNAs in exosome andnon-exosomal fraction of follicular fluid consisted of fullygrown or growing oocytes and to validate exosome medi-ated cell–cell communication between follicular cells. Forthis, follicles of 5–8mm diameter (n=120) were isolatedand individual COCs were subjected to brilliant cresyl blue(BCB) staining and classified as BCB+ (fully grown,n=60)and BCB−(growing,n=60) groups. The correspondingfollicular fluid, granulosa cells and theca cells were usedfor further molecular analysis. miRNAs isolated fromexosomal and nonexosomal portion of follicular fluidfrom the two categories was used for cDNA synthesis andsubsequent analysis using a human miRNA PCR array (with745 miRNA).own or growing oocytes and to validate exosome medi-ated cell–cell communication between follicular cells. Forthis, follicles of 5–8mm diameter (n=120) were isolatedand individual COCs were subjected to brilliant cresyl blue(BCB) staining and classified as BCB+ (fully grown,n=60)and BCB−(growing,n=60) groups. The correspondingfollicular fluid, granulosa cells and theca cells were usedfor further molecular analysis. miRNAs isolated fromexosomal and nonexosomal portion of follicular fluidfrom the two categories was used for cDNA synthesis andsubsequent analysis using a human miRNA PCR array (with745 miRNA).cell–cell communication between follicular cells. Forthis, follicles of 5–8mm diameter (n=120) were isolatedand individual COCs were subjected to brilliant cresyl blue(BCB) staining and classified as BCB+ (fully grown,n=60)and BCB−(growing,n=60) groups. The correspondingfollicular fluid, granulosa cells and theca cells were usedfor further molecular analysis. miRNAs isolated fromexosomal and nonexosomal portion of follicular fluidfrom the two categories was used for cDNA synthesis andsubsequent analysis using a human miRNA PCR array (with745 miRNA).</p