摘要
【目的】研究生长分化因子9(growth differentiation facter9,GDF9)的潜在靶蛋白Claudin-11在成年羊驼卵巢中的表达与定位,探索Claudin-11对促进卵泡发育、调节排卵的作用机理。【方法】以成年雌性羊驼为研究对象,用RT-PCR技术分析潜在靶蛋白Claudin-11在成年羊驼卵巢中的表达,并使用实时荧光定量PCR技术分析其相对表达量,采用免疫组织化学法和western blotting法对Claudin-11在成年羊驼卵巢中的表达进行定位分析。【结果】荧光定量PCR结果显示,Claudin-11在羊驼卵巢的初级卵泡中弱表达于次级卵泡;westernblotting结果显示,多克隆兔抗Claudin-11抗体与成年羊驼卵巢组织粗提取物可以发生特异性的免疫反应;免疫组织化学试验结果显示,Claudin-11在羊驼卵巢的原始卵泡和次级卵泡中弱表达于卵泡细胞周围的扁平或立方形卵泡细胞的胞质,而在次级卵泡中Claudin-11在最内层颗粒细胞的胞质中进行表达。【结论】Claudin-11在成年羊驼卵巢组织中表达且存在特异性的细胞定位。
【Objective】To explore the mechanism of Claudin-11 in promoting the follicular development and regulating the ovulation,an experiment was carried out to investigate the expression and localization of Claudin-11 in adult alpaca ovary.【Method】 Using the adult female alpaca as the object of study,the expression of Claudin-11 in the ovary was analyzed by RT-PCR and real-time PCR.Immunohistochemistry and western blotting were used to explore the localization of Claudin-11 in alpaca ovary.【Result】The expression of Claudin-11 in ovarian primary follicle was weaker than in the secondary follicle.Polyclonal antibody happened specific immune response with the crude extract from the ovarian.The expression of Claudin-11 in ovarian primary follicle and secondary follicle was weaker than that in flat or cube follicle cell cytoplasm.Claudin-11 was mainly located in the granulosa cell of the secondary follicle endothecium.【Conclusion】The results showed that Claudin-11 expressed in adult alpaca ovary,and there was a specific cellular localization.
出处
《中国农业科学》
CAS
CSCD
北大核心
2013年第10期2103-2109,共7页
Scientia Agricultura Sinica
基金
农业部"948"项目(2011-Z33)
农业公益性行业项目(201303119)
山西省自然科学基金(2012011046-6)
