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The global effect of follicle-stimulating hormone and tumour necrosis factor ? on gene expression in cultured bovine ovarian granulosa cells.

ABSTRACT: Oocytes mature in ovarian follicles surrounded by granulosa cells. During follicle growth, granulosa cells replicate and secrete hormones, particularly steroids close to ovulation. However, most follicles cease growing and undergo atresia or regression instead of ovulating. To investigate the effects of stimulatory (follicle-stimulating hormone; FSH) and inhibitory (tumour necrosis factor alpha; TNF?) factors on the granulosa cell transcriptome, bovine ovaries were obtained from a local abattoir and pools of granulosa cells were cultured in vitro for six days under defined serum-free conditions with treatments present on days 3-6. Initially dose-response experiments (n = 4) were performed to determine the optimal concentrations of FSH (0.33 ng/ml) and TNF? (10 ng/ml) to be used for the microarray experiments. For array experiments cells were cultured under control conditions, with FSH, with TNF?, or with FSH plus TNF? (n = 4 per group) and RNA was harvested for microarray analyses.Statistical analysis showed primary clustering of the arrays into two groups, control/FSH and TNF?/TNF? plus FSH. The effect of TNF? on gene expression dominated that of FSH, with substantially more genes differentially regulated, and the pathways and genes regulated by TNF? being similar to those of FSH plus TNF? treatment. TNF? treatment reduced the endocrine activity of granulosa cells with reductions in expression of FST, INHA, INBA and AMH. The top-ranked canonical pathways and GO biological terms for the TNF? treatments included antigen presentation, inflammatory response and other pathways indicative of innate immune function and fibrosis. The two most significant networks also reflect this, containing molecules which are present in the canonical pathways of hepatic fibrosis/hepatic stellate cell activation and transforming growth factor ? signalling, and these were up regulated. Upstream regulator analyses also predicted TNF, interferons ? and ?1 and interleukin 1?.In vitro, the transcriptome of granulosa cells responded minimally to FSH compared with the response to TNF?. The response to TNF? indicated an active process akin to tissue remodelling as would occur upon atresia. Additionally there was reduction in endocrine function and induction of an inflammatory response to TNF? that displays features similar to immune cells.

SUBMITTER: Glister C

PROVIDER: S-EPMC3906957 | biostudies-literature | 2014 Jan

REPOSITORIES: biostudies-literature

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The global effect of follicle-stimulating hormone and tumour necrosis factor α on gene expression in cultured bovine ovarian granulosa cells.

Glister Claire C Hatzirodos Nicholas N Hummitzsch Katja K Knight Philip G PG Rodgers Raymond J RJ

BMC genomics 20140128

<h4>Background</h4>Oocytes mature in ovarian follicles surrounded by granulosa cells. During follicle growth, granulosa cells replicate and secrete hormones, particularly steroids close to ovulation. However, most follicles cease growing and undergo atresia or regression instead of ovulating. To investigate the effects of stimulatory (follicle-stimulating hormone; FSH) and inhibitory (tumour necrosis factor alpha; TNFα) factors on the granulosa cell transcriptome, bovine ovaries were obtained fr ...[more]

PMID: 24467805

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Project description:BACKGROUND:Heifers not used as breeding stock are often implanted with steroids to increase growth efficiency thereby altering hormone profiles and potentially changing the environment in which ovarian follicles develop. Because bovine granulosa cell culture is a commonly used technique and often bovine ovaries are collected from abattoirs with no record of implant status, the objective of this study was to determine if the presence of an implant during bovine granulosa cell development impacts follicle stimulating hormone-regulated steroidogenic enzyme expression. Paired ovaries were collected from 16 feedlot heifers subjected to 1 of 3 treatments: non-implanted (n?=?5), Revalor 200 for 28 d (n?=?5), or Revalor 200 for 84 d (n?=?6). Small follicle (1 to 5 mm) granulosa cells were isolated from each pair and incubated with phosphate buffered saline (n?=?16) or 100 ng/mL follicle stimulating hormone (n?=?16) for 24 h. RESULTS:Granulosa cells of implanted heifers treated with follicle stimulating hormone produced medium concentrations of progesterone similar (P?=?0.22) to non-implanted heifers, while medium estradiol concentrations were increased (P?<?0.10) at 28 and 84 d compared to non-implanted heifers indicating efficacy of treatment. Additionally, real-time PCR analysis in response to follicle stimulating hormone treatment demonstrated a decrease in steroidogenic acute regulatory protein (P?=?0.05) mRNA expression in heifers implanted for 84 d and an increase in P450 side chain cleavage mRNA in granulosa cells of heifers implanted for 28 (P?<?0.10) or 84 d (P?<?0.05) compared to non-implanted females. However, no difference in expression of 3-beta-hydroxysteroid dehydrogenase (P?=?0.57) and aromatase (P?=?0.23) were demonstrated in implanted or non-implanted heifers. CONCLUSIONS:These results indicate follicles which develop in the presence of high concentrations of androgenic and estrogenic steroids via an implant tend to demonstrate an altered capacity to respond to follicle stimulating hormone stimulation. Thus, efforts should be made to avoid the use of implanted heifers to study steroidogenesis in small follicle granulosa cell culture systems.

Dataset Information

The global effect of follicle-stimulating hormone and tumour necrosis factor ? on gene expression in cultured bovine ovarian granulosa cells.

Publications

The global effect of follicle-stimulating hormone and tumour necrosis factor α on gene expression in cultured bovine ovarian granulosa cells.

Similar Datasets

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