Ontogeny of follicle-stimulating hormone receptor gene expression in isolated human ovarian follicles
Ontogeny of follicle-stimulating hormone receptor gene expression in isolated human ovarian follicles
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DOI：
10.1210/jc.82.11.3748
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发表时间：
1997-11-01
影响因子：
5.8
通讯作者：
Gosden, RG
中科院分区：
文献类型：
--
作者：
Oktay, K;Briggs, D;Gosden, RG
FSH刺激有腔卵泡生长，但其在早期阶段的作用（如果有的话）尚不清楚。我们的目的是确定卵泡阶段的FSH受体（FSHr）基因首次表达。我们使用基于PCR的策略来分析从人卵巢分离后从原始到多层阶段的单个卵泡。卵巢组织来自11名妇女（年龄范围，25-33岁）接受择期剖宫产。组织在含有1%胶原酶的培养基中部分解聚，并手动解剖卵泡，使其不含基质。显微镜下确认卵泡阶段为原始（不生长）、初级（1层立方颗粒细胞）或具有2层或更多层颗粒细胞。直肌和基质组织用作阴性对照。将每个卵泡的信使核糖核酸（mRNA）逆转录，并使用FSHr和肌动蛋白的引物通过巢式PCR扩增所得cDNA。9个原始卵泡均不表达FSHr mRNA。33%的初级卵泡和2层卵泡FSHr mRNA阳性（分别为4/12和3/9），100%（n = 4）的多层卵泡。FSH在生长卵泡和原始卵泡中的表达差异有统计学意义。这项研究首次表明，FSHr基因的转录开始于卵泡生长的最早阶段，并表明FSH可能在腔前卵泡发育中具有迄今为止未被怀疑的生理作用。此外，本研究还证明了研究人类卵泡发生过程中其他基因表达的实际可行性。
FSH stimulates antral follicles to grow, but its role in earlier stages, if any, is obscure. Our aim was to determine the follicle stage at which the FSH receptor (FSHr) gene is first expressed. We used a PCR-based strategy to analyze single follicles ranging from primordial to multilaminar stages after isolation from human ovaries. Ovarian tissue was obtained from 11 women (age range, 25-33 yr) undergoing elective cesarean section. Tissue was partially disaggregated in medium containing 1% collagenase, and follicles were manually dissected free of stroma. Follicle stages were confirmed microscopically as primordial (nongrowing), primary (1 layer of cuboidal granulosa cells), or having 2 or more layers of granulosa cells. Rectus muscle and stromal tissue were used as negative controls. Messenger ribonucleic acid (mRNA) from each follicle was reverse transcribed, and the resulting cDNA was amplified by nested PCR using primers for FSHr and actin. None of the 9 primordial follicles expressed FSHr mRNA. Thirty-three percent of the primary and 2-layer follicles were positive for FSHr mRNA (4 of 12 and 3 of 9, respectively), as were 100% (n = 4) of the multilaminar follicles. The difference in FSH expression between the growing and primordial follicles was significant. This study shows for the first time that transcription of the FSHr gene begins at the earliest stages of follicular growth and indicates that FSH may have a hitherto unsuspected physiological role in preantral follicle development. In addition, this study demonstrates the practical feasibility of investigating the expression of other genes during human folliculogenesis.