FGF1 induced Sprouty genes expression by RTK/ERK pathway in bovine ovarian granulosa cells | FGF1对牛卵巢颗粒细胞Spry基因表达的影响

Chinese. 研究体外培养牛卵巢颗粒细胞中Sprouty基因（Spry）的表达，探讨成纤维细胞生长因子1（FGF1）对Spry基因表达的影响。从屠宰场收集牛卵巢，剪下直径2～5 mm的卵泡，机械破碎卵泡后收集颗粒细胞进行体外培养，在培养的第5天用FGF1和PD98059处理培养的颗粒细胞，然后用Trizol提取细胞总RNA，利用牛特异性引物进行实时定量PCR检测，用ΔΔCt方法计算Spry基因的相对表达量。牛卵巢颗粒细胞可以表达Spry基因；Spry基因的表达与FGF1呈现剂量依赖关系，Spry2和Spry4的表达量随FGF1作用时间的延长呈现出先上升后下降的变化趋势；FGF1信号通过RTK/ERK通道对Spry基因的表达产生刺激作用。在体外培养的牛卵巢颗粒细胞中，FGF1信号通过RTK/ERK通道刺激牛Spry基因表达。

English. The aim of the present experiments was to study the expression of Sprouty (Spry) genes in bovine ovarian granulosa cells, and the effects of fibroblast growth factor-1 (FGF1) signaling on Spry genes expression. Bovine ovaries were collected from a local abattoir, and follicles diameter between 2-5 mm were dissected to isolate granulosa cells by mechanical way, and the granuloca cells were cultured for 5 days in vitro. On day 5, granulosa cells were treated with FGF1 and PD98059, and total RNA was extracted from the culutred cells with Trizol. Real-time PCR was used to determine the expression of Spry genes using bovine specific primers and to calculate the relative expression of Spry genes withΔΔCt. ① Spry genes were expressed in bovine ovarian granulosa cells; ② Dose-dependent relationship was observed between the expression of Spry genes and FGF1 signaling. The expression of Spry2 and Spry4 increased significantly at first and then decreased with the treating of FGF1, and FGF1 signaling induced Spry genes expression in bovine ovarian granulosa cells by RTK/ERK pathway. FGF1 induced the expression of Spry genes in bovine ovarian granulosa cells cultured in vitro by RTK/ERK pathway.