文献分享 | ECHO显微镜在抗穆勒氏管激素研究中的应用

在2017年12月27日，由Jing Xu，Fuhua Xu，Maralee S Lawson，Olena Y Tkachenko等作者在Biology of Reproduction中发表一篇了《Anti-Mullerian hormone is a survival factor and promotes the growth of rhesusmacaque preantral follicles during matrix-free culture》的文章。

文章使用Revolve正倒置一体显微镜获得了如下结果:

该图中B-E是通过Revolve正倒置一体显微镜成像得到的结果 ↑

该图中A-B、C-F是通过Revolve正倒置一体显微镜成像得到的结果 ↑

中文摘要：

抗穆勒氏管激素（AMH）在卵泡发育过程起着至关重要的作用，其局部作用于卵泡生长过程中的动态分泌有关。目前不同物种的AMH在腔内卵泡生长和功能的影响已有所验证，但对腔前卵泡发育的作用仍待进一步验证。因此，我们通过实验研究AMH在腔前卵泡发育过程中所起到的作用。利用AMH-shRNA重组腺病毒载体降低猕猴卵泡中AMH基因的表达。将次生卵泡从成年猕猴卵巢中分离出来后用超低附着培养皿单独培养，且培养基中加入促卵泡激素和胰岛素，培养5周。将卵泡随机分配给治疗组，分别为a.对照组；b.非靶向对照组shRNA-vector；c.AMH shRNA-vector；d.AMH shRNA-vector + 重组人源AMH；e.重组人源AMH。评估卵泡的存活和生长情况，并分析培养基中的类固醇激素和旁分泌因子的浓度。在体内研究中，将非靶向对照shRNA-vecotor和AMH shRNA-vector分别注入猕猴的卵巢，术后9天对卵巢进行形态学及免疫组化检测。结果发现，在非人灵长类动物中，AMH表达下调抑制腔前卵泡的存活和生长。体外实验发现，在培养基中添加AMH则可促进腔前卵泡生长发育至小腔窦期，类固醇激素和旁分泌因子表达增加，并促进卵母细胞成熟。这些数据表明，AMH是一个关键的卵泡旁分泌/自分泌因子，促进灵长类的腔前卵泡存活和生长。

英文摘要：

Anti-Mullerian 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 incor- porated 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 hor- mone 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 + recombi-nant 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 for morphology and immunohistochemistry assessment. Decreased AMH expression reduced preantral follicle survival and growth in non- human primates. Supplemental AMH treatment in the culture media promoted preantral follicle growth to the small antral stage in vitro with increased steroid hormone and paracrine factor production, as well as oocyte maturation. These data demonstrate that AMH is a critical follicular paracrine/autocrine factor positively impacting preantral follicle survival and growth in primates.

ECHO Revolve正倒置一体显微镜展现了其非凡的灵活性，可以轻松地实现正置和倒置显微镜转换，创新性地把正倒置显微镜合二为一，开启了显微镜Hybrid时代。

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ECHO Revolve正倒置一体显微镜兼具正置和倒置显微镜的优势，方便小巧，一机多能，可以非常便利地通过旋转实现正倒置配置切换；以iPad桌面替代目镜设计, 获得的视觉体验；同时基于iOS的Echo app，使软件操作更人性化。