Project description:The study aimed to uncover differential expression pattern of regulatory microRNAs in bovine granulosa cells derived from preovulatory dominant and subordinate follicles.
Project description:Bovine granulosa cells harvested from cohort follicles at 1.3 days after wave emergence, and from dominant and subordinate follicles at 2.6 days after wave emergence. Total RNA extracted from granulosa cells of 4 cohort, 4 dominant and 4 subordinate follicles was pooled for generation of each respective SAGE library. Keywords: other
Project description:Bovine granulosa cells harvested from cohort follicles at 1.3 days after wave emergence, and from dominant and subordinate follicles at 2.6 days after wave emergence. Total RNA extracted from granulosa cells of 4 cohort, 4 dominant and 4 subordinate follicles was pooled for generation of each respective SAGE library. Keywords: other
Project description:Development of ovarian follicles is controlled at the molecular level by several gene products whose precise expression leads to regression or ovulation of follicles. MicroRNAs (miRNAs) are small non-coding RNAs that regulate gene expression through sequence-specific base pairing with target messenger RNAs (mRNAs) causing translation repression or mRNA degradation. The aim of this study was to identify miRNAs expressed in ovarian follicles, localize their expression within the theca and/or granulosa layers and their putative target genes/pathways that are involved in bovine ovarian follicle development. By using miRCURY microarray (Exiqon) we identified 14 and 49 differentially expressed miRNAs (P < 0.01) between dominant and subordinate follicles in theca and granulosa cells, respectively. The expression levels of four selected miRNAs were confirmed by qRT-PCR. To identify target prediction and pathways of differentially expressed miRNAs Union of Genes option in DIANA miRPath v.2.0 software was used. The predicted targets for these miRNAs were enriched for pathways involving oocyte meiosis, Wnt, TGF-beta, ErbB, Insulin, P13K-Akt and MAPK signaling pathways. This study identified differentially expressed miRNAs in the theca and granulosa cells of dominant and subordinate follicles, and clearly implicates them in having important roles in regulating known molecular pathways that determine the fate of ovarian follicle development.
Project description:The expression pattern of microRNAs in granulosa cells of subordinate and dominant follicles during the early luteal phase of bovine estrous cycle
Project description:The objective of the study was to determine how maternal age influences the transcriptome of the dominant follicle during the preovulatory period. We tested the hypotheses that delayed ovulation in aged cows is associated with 1) altered gene expression of granulosa cells of preovulatory follicles (24 h after LH treatment) and 2) decreased synthesis of progesterone by granulosa cells of the preovulatory follicle. Granulosa cells of preovulatory follicles obtained 24 h after LH treatment from aged Hereford cows (19.0 ±2.5 years; n=3) were compared to those from young cows (9.0 ± 0.6 years; n=3) using bovine specific microarrays (EmbryoGENE-EMBV3; GPL13226). Results were confirmed by RT-qPCR. A total of 1340 genes or gene isoforms were expressed differentially (â¥2-fold change; p ⤠0.05) in aged cows vs. young cows (daughters of aged cows). In conclusion, transcriptome analysis of granulosa cells from aged cows revealed a delayed or suboptimal response to the preovulatory LH stimulus, represented by delayed cellular differentiation, luteinization and progesterone synthesis. Granulosa cells of the dominant preovulatory follicle 24 h after LH treatment were compared between aged vs.young cows. Three biological replicates (each composed of one aged and one young cow). 3 Three technical replicate (dye swap). One biological or technical replicate per array.