title: Constitutive and follicle-stimulating hormone-induced action of somatostatin receptor-2 on regulation of apoptosis and steroidogenesis in bovine granulosa cells.
abstract: In the present study, we employed primary bovine culture of granulosa cells (GCs) as a cellular model to study the potential involvement of somatostatin receptor 2 (SSTR2) in ovarian function. The results showed that bovine GCs expressed SST2 receptor and further found that SSTR2 was possibly regulated by follicle-stimulating hormone (FSH), as a significant increase in protein level of SSTR2 was observed in FSH-treated GCs. For further analysis, endogenous SSTR2 expression was disrupted using small inhibitory RNA (siRNA) and the efficacy of differential silencing of endogenous SSTR2 expression was measured both at transcriptional and translational levels. Transient blockage of SSTR2 evidenced its constitutive action on GCs, as it significantly increased level of cAMP (2.4-folds) and basal progesterone production (∼2-fold, P<0.05) with significant increase (P<0.05) in mRNA levels of StAR and P450ssc without altering estradiol concentration and aromatase mRNA expression. Furthermore, silencing of SSTR2 reduced GCs apoptosis (52.5%, P<0.05) and increased cell proliferation, which was further corroborated by up-regulation in protein expressions of B-cell leukemia/lymphoma 2 (Bcl-2), inhibition of caspase3 and mRNA level of bcl2-associated-X protein (Bax). These results provide evidence that SSTR2 subtype controls GCs apoptosis, proliferation and hormonal secretions through selective constitutive action, independently of somatostatin (SST). Given the local inhibitory actions of SSTR2 on the gonads, we further found that apoptosis in ssRNAi-2 transfected cells decreased (6.8% vs 1.9%, P<0.05) more strongly on FSH treatment. Apoptotic protein expressions and steroid hormone mRNA levels were correlated with a relative decrease in apoptosis and increase in progesterone production. Our results suggest that SSTR2 may play a crucial role as a local inhibitor of FSH action on GCs apoptosis and steroidogenesis.
authors: Key Laboratory of Agricultural Animal Genetics, Breeding and Reproduction, Education Ministry of China, Huazhong Agricultural University, Wuhan, Hubei 430070, People's Republic of China.
authors: Hasan
authors: H
authors: Riaz
authors: Key Laboratory of Agricultural Animal Genetics, Breeding and Reproduction, Education Ministry of China, Huazhong Agricultural University, Wuhan, Hubei 430070, People's Republic of China.
authors: Ping
authors: P
authors: Dong
authors: College of Animal Sciences, Department of Clinical Veterinary Medicine, Huazhong Agricultural University, Wuhan, Hubei 430070, People's Republic of China.
authors: Muhammad
authors: M
authors: Shahzad
authors: Key Laboratory of Agricultural Animal Genetics, Breeding and Reproduction, Education Ministry of China, Huazhong Agricultural University, Wuhan, Hubei 430070, People's Republic of China. Electronic address: yangliguo2006@yahoo.com.cn.
authors: Liguo
authors: L
authors: Yang
mesh_terms: D000818:Animals; D017209:Apoptosis; D002417:Cattle; D049109:Cell Proliferation; D002478:Cells, Cultured; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics / Q000378:metabolism; D005260:Female; D005640:Follicle Stimulating Hormone / Q000502:physiology*; D015870:Gene Expression; D006107:Granulosa Cells / Q000502:physiology*; D019950:Mitogen-Activated Protein Kinase 1 / Q000378:metabolism; D048052:Mitogen-Activated Protein Kinase 3 / Q000378:metabolism; D011374:Progesterone / Q000096:biosynthesis*; D017481:Receptors, Somatostatin / Q000235:genetics / Q000378:metabolism*; D015533:Transcriptional Activation
chemical_list: D017481:Receptors, Somatostatin; D011374:Progesterone; D005640:Follicle Stimulating Hormone; D003577:Cytochrome P-450 Enzyme System; D019950:Mitogen-Activated Protein Kinase 1; D048052:Mitogen-Activated Protein Kinase 3
keywords: Apoptosis; RNAi; SSTR2; bcl2