Unknown,Transcriptomics,Genomics,Proteomics

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Transcriptome profiling of dominant follicle development in the equine ovary

ABSTRACT: The Objective of this study was to establish global gene expression profiles associated with different stages of dominant follicle development in the horse. This was done by collecting ovaries at different stages of an ovulatory follicular wave, when the dominant follicle reached 22 mm (early dominant, ED), 33 mm (late dominant, LD) and 34 h after an i.v. injection of crude equine gonadotropins (CEG; 15 mg i.v.) when the largest follicle reached > 33 mm (preovulatory stage, PO). RNA was then separately collected from granulosa cells and theca-rich follicular wall fractions and was hybridized to the Agilent Horse Gene Expression Microarray. Gene expression was compared between ED and LD and between LD and PO stages within each of granulosa cells and theca walls. Significantly larger numbers of genes were differentially expressed in granulosa cells than in theca walls throughout development, and between LD and PO follicles than between ED and LF follicles. The most salient features were the downregulation of cell cycle genes in granulosa cells during both the ED-LD and LD-PO transitions and the upregulation of genes associated with inflammation, immunity, extracellular matrix remodelling and protection aganints toxic insult in both GCs and TCs, particularly during the LD-PO transition. Five to nine biological replicates per cell type and developmental stage (total of 34 samples) were used in a single dye experiment. Samples were distributed among slides so that each experimental groupeach was represented at least once in each slide. For each gene, mean normalized intensities (n= 5 biological replicates/group) were compared between follicle stages (ED-LD and LD-PO) within each cell type (GC and TC).

ORGANISM(S): Equus caballus

SUBMITTER: Xavier Donadeu

PROVIDER: E-GEOD-52109 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:The molecular mechanisms that regulate the pivotal transformation processes observed in the follicular wall following the pre-ovulatory LH-surge, are still not established, particularly for cells of the thecal layer. To elucidate thecal and granulosa cell type-specific biological functions and signaling pathways, large dominant bovine follicles were collected before and 21 hrs after an exogenous GnRH induced LH surge. Because LH receptor density varies within the granulosa cell populations, antral granulosa (aGC; those aspirated by follicular puncture) and membrane associated granulosa (mGC; those scraped from the follicular wall) were compared to thecal cell expression profiles determined by mRNA microarrays. Thecal cell gene expression was less affected in the peri-ovulatory follicle when compared to granulosa cells, as evidenced by only 2% versus 25% of the ~11,000 genes expressed changing in response to the LH surge, respectively. The majority of the 203 LH-regulated thecal genes were also LH regulated in granulosa cells, leaving a total of 58 genes as LH-regulated theca cell specific genes. Most of the 58 genes (i.e., 74%) thecal specific genes including several known thecal markers (CYP17A1, NR5A1) were downregulated, while most genes identified are new to theca. Many of the newly identified upregulated thecal genes (e.g., PTX3, RND3, PPP4R4) were also upregulated in granulosa. Minimal expression differences were observed between aGC and mGC, however, transcripts encoding extracellular proteins (NID2) and matrix modulators (ADAMTS1, SASH1) predominated these differences. We also identified large numbers of unknown LH-regulated granulosa cell genes and discuss their putative roles in ovarian function. The single dominant ovarian follicle was collected from each cow before the LH surge or 22 hours after GnRH (used to induce LH surge). RNA was extracted from three independent cells within each follicle and there were hybridized on Affymetrix microarrays.

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Transcriptome profiling of dominant follicle development in the equine ovary

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