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首页> 美国卫生研究院文献>Biology of Reproduction >Anti-Müllerian hormone is a survival factor and promotes the growth of rhesusmacaque preantral follicles during matrix-free culture
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Anti-Müllerian hormone is a survival factor and promotes the growth of rhesusmacaque preantral follicles during matrix-free culture

机译：抗苗勒管激素是一种生存因子可促进恒河猴的生长猕猴无基质培养前卵泡

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Anti-Müllerian hormone (AMH) plays a key role during ovarian follicular development, with local actions associated with a dynamic secretion profile by growing follicles. While results for AMH effects on antral follicle growth and function are consistent among studies in various species, any effects on preantral follicle development remain controversial. Therefore, experiments were conducted to investigate the direct actions and role of AMH during follicle development at the preantral stage. Macaque-specific short-hairpin RNAs (shRNAs) targeting AMH mRNA were incorporated into adenoviral vectors to decrease AMH gene expression in rhesus macaque follicles. Secondary follicles were isolated from adult macaque ovaries and cultured individually in the ultra-low-attachment dish containing defined medium supplemented with follicle-stimulating hormone and insulin for 5 weeks. Follicles were randomly assigned to treatment groups: (a) control, (b) nontargeting control shRNA-vector, (c) AMH shRNA-vector, (d) AMH shRNA-vector + recombinant human AMH, and (e) recombinant human AMH. Follicle survival and growth were assessed. Culture media were analyzed for steroid hormone and paracrine factor concentrations. For in vivo study, the nontargeting control shRNA-vector and AMH shRNA-vector were injected into macaque ovaries. Ovaries were collected 9 days postinjection formorphology and immunohistochemistry assessment. Decreased AMH expressionreduced preantral follicle survival and growth in nonhuman primates. Supplemental AMHtreatment in the culture media promoted preantral follicle growth to the small antralstage in vitro with increased steroid hormone and paracrine factor production, as well asoocyte maturation. These data demonstrate that AMH is a critical follicularparacrine/autocrine factor positively impacting preantral follicle survival and growth inprimates.
机译：抗苗勒管激素（AMH）在卵巢卵泡发育过程中起关键作用，其局部作用与卵泡生长引起的动态分泌有关。尽管AMH对肛门卵泡生长和功能的影响的结果在不同物种的研究中是一致的，但对肛门前卵泡发育的任何影响仍存在争议。因此，进行了实验，以研究AMH在产前期卵泡发育过程中的直接作用和作用。将针对AMH mRNA的猕猴特异性短发夹RNA（shRNA）整合到腺病毒载体中，以减少猕猴卵泡中AMH基因的表达。从成年猕猴卵巢中分离出次级卵泡，并在超低附着培养皿中单独培养，该培养皿中含有确定的培养基，其中添加了刺激卵泡的激素和胰岛素，持续5周。卵泡被随机分为治疗组：（a）对照，（b）非靶向对照shRNA载体，（c）AMH shRNA载体，（d）AMH shRNA载体+重组人AMH和（e）重组人AMH。评估卵泡存活和生长。分析培养基中的类固醇激素和旁分泌因子浓度。为了进行体内研究，将非靶向对照shRNA-载体和AMH shRNA-载体注射到猕猴卵巢中。注射后9天收集卵巢形态学和免疫组化评估。 AMH表达降低降低非人类灵长类动物的窦前卵泡存活和生长。补充AMH培养基中的处理促进了窦前卵泡向小肛门的生长类固醇激素和旁分泌因子产生增加的体外阶段，以及卵母细胞成熟。这些数据表明AMH是关键的卵泡旁分泌/自分泌因子积极影响食管前卵泡的存活和生长灵长类动物。

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