Unknown,Transcriptomics,Genomics,Proteomics

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Transcriptome analysis of granulosa cells after conventional vs long FSH-induced superstimulation in cattle

ABSTRACT: Conventional Superstimulation (group 1) vs. Long Superstimulation (group 3) A genome-wide bovine oligo-microarray was used to compare the gene expression of granulosa cells collected from ovarian follicles after differing durations of the growing phase induced by exogenous FSH treatment. Cows were given a conventional (4-day) or long (7-day) superstimulatory treatment (25mg FSH im at 12-h intervals; n=6 per group), followed by prostaglandin treatment with last FSH and LH treatment 24 hr later. Granulosa cells were harvested 24 h after LH treatment. The expression of 416 genes was down-regulated and 615 genes was up-regulated in the long FSH group compared to the conventional FSH group. Quantification by RT-PCR of 7 genes (NTS, PTGS2, PTX3, RGS2, INHBA, CCND2 and LRP8) followed the same trend as in microarrays. Compared to the conventional group, the long FSH group responded to exogenous LH with down-regulation of mRNA for cyclinD2, lipoprotein receptorP8, CYP51A1, CYP19A1, CJA1, INHBA, SRPINE1, and up-regulation of Vannin, POSTN, GTPAse, Cysteine. Markers of fertility and follicle maturity were up-regulated in the long FSH group. We conclude that a prolonged FSH-induced growing phase is associated with transcriptomic characteristics of greater follicular maturity and may therefore be more appropriate for optimizing the superovulatory response and developmental competence of oocytes in cattle. straight comparison of Short Superstimulation group (the reference; group 1) versus a Long Superstimulation protocol ( the treatment; group 3) using 3 different animals (biological replicates) in each group and performed dye swap. For example on array 1: group 1 cow 1 versus group 3 cow 1 (cy3 vs cy5) and on array 4 is the dye swap group 3 cow 1 versus group 1 cow 1 (cy3 vs cy5).

ORGANISM(S): Bos taurus

SUBMITTER: Fernanda Dias

PROVIDER: E-GEOD-80289 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:The main objective of superovulatory treatment is to achieve multiple ovulations and produce multiple embryos. The technique is intended to rescue a cohort of follicles within a follicular wave that would otherwise regress. Superstimulory treatments change the intra-follicular and systemic hormonal milieu, but it is not clear how and to what extent treatment alters gene expression of follicular cells. In this study, a genome-wide bovine oligo-microarray was used to compare the gene expression of granulosa cells from post-LH preovulatory dominant follicle with those from follicles of the same status after superstimulatory treatment. Cows were allocated randomly to two groups (superstimulation group and control group, n=6 cows per group). A new follicular wave was induced by transvaginal ultrasound-guided ablation of follicles ?5 mm in diameter, and a progesterone-releasing device (CIDR) was placed in vagina. The superstimulation group was given eight doses of 25 mg FSH im at 12-h intervals starting from the day of wave emergence (Day 0), whereas the control group was not given any FSH treatment. Both groups were given prostaglandin F2? im twice, 12 h apart, on Day 3 and the CIDR was removed at the second injection; 25 mg pLH was given im 24 h after CIDR removal, and cows were ovariectomized 24 h later. Granulosa cells were collected for RNA extraction, amplification and microarray hybridization. To translate microarray results to a physiological context, a list of differentially expressed transcripts were biologically annotated. A total of 190 genes were down-regulated and 280 genes were up-regulated in superstimulated group when compared with the reference (non-superstimulated control) group. straight comparison of superstimulation group (the treatment; group 1) versus non-superstimulated ( the reference; group 4) using 3 different aninals (biological replicates) in each group and performed dye swap. For example on array 1: group1 cow 1 versus group 4 cow 1 (cy3 vs cy5) and on array 4 is the dey swap group 4 cow 1 versus group 1 cow 1 (cy3 vs cy5).

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Transcriptome analysis of granulosa cells after conventional vs long FSH-induced superstimulation in cattle

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