羔羊卵巢AMH的免疫组织化学分析  

Immunocytochemical Study of Anti-Müllerian Hormone in Ovarian Follicles from Lambs

作　　者：蒋香菊[1,2] 吴阳升 张利[1,2] 古丽米热.阿不都热依木 林嘉鹏 汪立芹 杨楠[2,3] 唐淑红[2,3] 黄俊成 

机构地区：[1]石河子大学动物科技学院,新疆石河子832000 [2]农业部草食家畜遗传育种与繁殖重点实验室,乌鲁木齐830000 [3]新疆农业大学,乌鲁木齐830052

出　　处：《西北农业学报》2014年第7期19-24,共6页Acta Agriculturae Boreali-occidentalis Sinica

基　　金：国家自然科学基金(U1203381);新疆维吾尔自治区公益性科研院所基本科研业务经费项目;国家重大转基因专项(2014ZX08008-003);新疆维吾尔自治区高技术项目(201111113);新疆维吾尔自治区科技支疆项目(201291147)

摘　　要：为探讨AMH在羔羊和成年羊卵巢中时间和空间表达变化,揭示卵泡生长发育调控的机制。收集1月龄羔羊和正常成年羊卵巢,采用HE染色观察卵巢的组织学特征,免疫组化法检测绵羊卵巢AMH蛋白表达分布,RT-PCR技术分析卵巢卵丘细胞中AMH基因的表达变化。结果显示,卵巢HE染色后,对外源激素处理有反应的卵巢存在大量的大直径卵泡,且无中等卵泡和小卵泡,也没有黄体化和排卵;对外源激素没有反应的卵巢仅存在原始卵泡和少量的初级卵泡,无有腔卵泡。免疫组化结果显示,AMH存在于生长期的腔前卵泡和小的有腔卵泡,以及卵丘细胞,大多原始卵泡无AMH的表达,而在原始卵泡向初级卵泡转变,外层的颗粒细胞由扁平向柱状转变时AMH有表达,颗粒细胞存在表达AMH和不表达AMH 2种类型。RT-PCR结果显示,羔羊卵丘细胞的表达量高于成年羊。AMH可能参与卵泡发育的启动、向促性腺激素依赖期的转变等过程的调控,其功能作用还有待于深入研究。To investigate the temporal and spatial expression patterns of AMH in lamb and ewe ovary. Histological and immunohistochemistry of AMH were analyzed on ovaries from one-month-old lambs and ewes. The mRNA expression of AMH on the cumulus cell from in vitro maturation （IVM） of cu- mulus-oocyte complex （COC） was detected using quantitative RT-PCR. The ovaries were over-stimu- lated in that all the surfaces were filled with large follicles without any small ones, and without lutein- ization or ovulation. In addtion, a large number of primordial follicles were present on the gonadotro- pin-unstimulated lamb＇s ovary. The immunoreactivity of AMH was limited to granulosa cells in growing follicles, and was localized in the granulosal cells of preantral and small antral follicles, aswell as in the cumulus oophorus, and no immunoreactivity was observed in most of the primordial fol- licles, except in transitory follicles contained an ooeyte surrounded by a single-layer mixture of flat- tened and cuboidal granulosal cells. The granulosal cell had been differentiated two types, expressing AMH and unexpressing AMH cell lines. The expression of AMH had no significant difference in the cumulus cell from IVM of COC between the one-month-old lamb and ewe. These observations suggest that AMH plays a key role in the modulation of the responsiveness of follicles progress through the gonadotropin-responsive to gonadotropin-dependent stage. A better understanding of AMH function is needed to uncover the mechanism which participates in managing the reserve of primordial and small growing follicles.

关 键 词：AMH 羔羊 卵巢 颗粒细胞 

分 类 号：S826[农业科学—畜牧学]