摘要
为揭示Wnt1和Frizzled(Fzd)2参与卵巢和卵泡发育的作用机制,选择12只3周龄小鼠,应用免疫组化研究Wnt1和Fzd2在卵巢组织内的定位,免疫共沉淀鉴定其蛋白间的配受体关系.结果表明:在健康初级、次级卵泡卵母细胞以及腔前和腔后卵泡近基膜颗粒细胞的胞质和胞膜中,Wnt1均表现较强的免疫染色,Fzd2在胞质内免疫染色;在卵泡腔周围颗粒细胞和透明带周围卵丘细胞中Wnt1免疫染色弱;从卵巢中心扩展或环绕生长卵泡的条形基质细胞中Wnt1和Fzd2免疫染色阳性,间质腺存在免疫染色细胞;在闭锁卵泡卵母细胞中Wnt1和Fzd2呈非均匀强染色;在卵母细胞裂解液的Wnt1或Fzd2免疫沉淀物的电泳图谱中,分别同时存在Fzd2和Wnt1蛋白条带.表明Wnt1定位于卵母细胞和近基膜颗粒细胞的胞质和胞膜中,Fzd2定位于胞质内;在卵巢基质细胞、卵泡膜细胞和间质腺细胞中存在Wnt1和Fzd2分布;Wnt1和Fzd2之间可能为配体-受体关系.
In order to reveal the mechanism of Wnt1 and Frizzled（Fzd） 2 in the regulation of ovarian and follicular development,twelve 3-week old mice were used to investigate the localization of Wnt1 and Fzd2 in the ovary by immunohistochemistry（IHC） and their ligand-receptor relationship by co-immunoprecipitation（Co-IP）.The results showed that a strong immunostaining of Wnt1 was found in the cytoplasm and on the member of oocytes of health primary and secondary follicles,as well as that in the granular cells near basal lamina of preantral and antral follicles,however Fzd2 staining was found only in the cytoplasm.There was weakly immunostaining of Wnt1 in granular cells around antrum and cumulus oophorus around zona pellucida.The elongated stromal cells round developing follicles or branched from ovarian cores were immunostained by Wnt1 and Fzd2.Furthermore,some scattered cells were also immunostained in the interstitial gland,while in the oocytes of follicular atresia,a strong and asymmetrical immunostaining was also found.Co-IP analysis showed that there was a direct interaction between Fzd2 and Wnt1 in the ovarian oocytes.Together,the results over clearly demonstrated the expression and localization of Wnt1 and Fzd2 in a cell-specific manner and their ligand-receptor relation,implying the important roles during ovarian follicular development.
出处
《浙江大学学报（农业与生命科学版）》
CAS
CSCD
北大核心
2011年第3期289-294,共6页
Journal of Zhejiang University:Agriculture and Life Sciences
基金
安徽科技学院引进和稳定人才科研资助项目(09RC10)
