Onset and Heterogeneity of Responsiveness to FSH in Mouse Preantral Follicles in Culture.
Onset and Heterogeneity of Responsiveness to FSH in Mouse Preantral Follicles in Culture.
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DOI：
10.1210/en.2016-1435
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发表时间：
2017-01-01
期刊：
影响因子：
4.8
通讯作者：
Franks S
中科院分区：
文献类型：
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作者：
Hardy K;Fenwick M;Mora J;Laird M;Thomson K;Franks S
促卵泡激素（follicle-stimulating hormone， FSH）在卵巢窦腔卵泡的正常发育和功能中所起的强制性作用已被公认，但其在窦腔前生长中的作用尚不清楚。本研究的具体目的是研究在培养过程中，不断增大的小鼠腔前卵泡对卵泡刺激素的反应，特别关注其生长速度和基因表达。从产后12 ~ 16天的C57BL/6小鼠卵巢中机械分离出腔前卵泡，并将单个卵泡单独在培养基中培养96小时（n = 511）或与重组人卵泡刺激素10 ng/mL （n = 546）。根据6个地层的初始卵泡直径进行分组，范围为<100 ~ >140 μm。在没有FSH的情况下，各种大小的卵泡均能生长（P < 0.01，配对t检验）。在FSH浓度为10 ng/mL的情况下，所有卵泡的生长速度都更快（P < 0.0001），但卵泡越大，对FSH的反应变化最大。即使最小的卵泡也表达FSH受体信使RNA （mRNA）。蛋白激酶A （PKA）抑制剂KT5720可抑制fsh诱导的卵泡生长，提示PKA途径与fsh诱导的卵泡生长有关。体外FSH处理后，FSH受体mRNA（定量聚合酶链反应测定）和Amh （P < 0.01）均降低（P < 0.01），而StAR （P < 0.0001）和甾体生成酶Cyp11a1 （P < 0.01）和Cyp19 （P < 0.0001）的表达均升高。这些结果显示卵泡对卵泡刺激素的不同反应取决于初始卵泡大小，较小的卵泡对卵泡刺激素的依赖程度低于较大的卵泡。这些发现强烈提示卵泡刺激素在腔前卵泡生长和功能中具有生理作用。
The obligatory role of follicle-stimulating hormone (FSH) in normal development and function of ovarian antral follicles is well recognized, but its function in preantral growth is less clear. The specific objective of this study was to investigate the response, in culture, to FSH of mouse preantral follicles of increasing size, focusing particularly on growth rate and gene expression. Preantral follicles were mechanically isolated from ovaries of C57BL/6 mice, 12 to 16 days postpartum, and single follicles cultured for up to 96 hours in medium alone (n = 511) or with recombinant human FSH 10 ng/mL (n = 546). Data were grouped according to initial follicle diameter in 6 strata ranging from <100 to >140 μm. Follicles of all sizes grew in the absence of FSH (P < 0.01, paired t test). All follicles grew at a faster rate (P < 0.0001) in the presence of 10 ng/mL FSH but larger follicles showed the greatest change in response to FSH. Even the smallest follicles expressed FSH receptor messenger RNA (mRNA). FSH-induced growth was inhibited by KT5720, an inhibitor of protein kinase A (PKA), implicating the PKA pathway in FSH-induced follicle growth. In response to FSH in vitro, FSH receptor mRNA (measured by quantitative polymerase chain reaction) was reduced (P < 0.01), as was Amh (P < 0.01), whereas expression of StAR (P < 0.0001) and the steroidogenic enzymes Cyp11a1 (P < 0.01) and Cyp19 (P < 0.0001) was increased. These results show heterogeneous responses to FSH according to initial follicle size, smaller follicles being less FSH dependent than larger preantral follicles. These findings strongly suggest that FSH has a physiological role in preantral follicle growth and function.