Department of Obstetrics, Affiliated Longhua People's Hospital, Southern Medical University (Longhua People's Hospital), Shenzhen, Guangdong 518109, PR China.
College of Animal Science and Technology, Northwest A&F University, Yangling, Shaanxi 712100, PR China.
Domest Anim Endocrinol. 2024 Oct;89:106859. doi: 10.1016/j.domaniend.2024.106859. Epub 2024 May 23.
GATA4 plays a pivotal role in the reproductive processes of mammals. However, the research on GATA4 in goat ovary is limited. This study aimed to study the expression and function of GATA4 in goat ovary. Utilizing real-time PCR and western blot analysis, we studied the expression and regulatory mechanisms of GATA4 in goat ovary and granulosa cells (GCs). We found that GATA4 was expressed in all follicle types in the goat ovary, with significantly higher levels in GCs of larger follicles (>3 mm) compared to those in smaller follicles (<3 mm). Additionally, we demonstrated that human chorionic gonadotrophin (hCG) induced GATA4 mRNA expression via the activation of PKA, MEK, p38 MAPK, PKC, and PI3K pathways in vitro. Our study also showed that hCG suppressed the levels of miR-200b and miR-429, which in turn directly target GATA4, thereby modulating the basal and hCG-induced expression of GATA4. Functionally, we examined the effect of siRNA-mediated GATA4 knockdown on cell proliferation and hormone secretion in goat GCs. Our results revealed that knockdown of GATA4, miR-200b, and miR-429 suppressed cell proliferation. Moreover, knockdown of GATA4 decreased estradiol and progesterone production by inhibiting the promoter activities of CYP11A1, CYP19A1, HSD3B, and StAR. Collectively, our findings suggest a critical involvement of GATA4 in regulating goat GC survival and steroidogenesis.
GATA4 在哺乳动物的生殖过程中起着关键作用。然而,关于山羊卵巢中 GATA4 的研究有限。本研究旨在研究 GATA4 在山羊卵巢中的表达和功能。利用实时 PCR 和 Western blot 分析,我们研究了 GATA4 在山羊卵巢和颗粒细胞(GCs)中的表达和调控机制。我们发现 GATA4 在山羊卵巢的所有卵泡类型中均有表达,在较大卵泡(>3mm)的 GC 中表达水平明显高于较小卵泡(<3mm)。此外,我们证明人绒毛膜促性腺激素(hCG)通过体外激活 PKA、MEK、p38 MAPK、PKC 和 PI3K 通路诱导 GATA4 mRNA 表达。我们的研究还表明,hCG 抑制了 miR-200b 和 miR-429 的水平,miR-200b 和 miR-429 可以直接靶向 GATA4,从而调节 GATA4 的基础表达和 hCG 诱导的表达。功能上,我们检查了 siRNA 介导的 GATA4 敲低对山羊 GCs 增殖和激素分泌的影响。结果表明,GATA4、miR-200b 和 miR-429 的敲低抑制了细胞增殖。此外,通过抑制 CYP11A1、CYP19A1、HSD3B 和 StAR 的启动子活性,GATA4 的敲低降低了雌二醇和孕酮的产生。综上所述,我们的研究结果表明 GATA4 参与调节山羊 GC 的存活和类固醇生成。