Rannikki A S, Zhang F P, Huhtaniemi I T
Department of Physiology, University of Turku, Finland.
Mol Cell Endocrinol. 1995 Feb;107(2):199-208. doi: 10.1016/0303-7207(94)03444-x.
The ontogeny of the follicle-stimulating hormone (FSH) receptor (R) gene expression was studied in the rat testis and ovary between day 12.5 or 14.5 of fetal life (f), respectively, and adulthood. In Northern blots hydbridized with a cRNA probe corresponding to a part of the extracellular domain of the FSHR, specific hybridization to testicular RNA was detected from day f18.5, and to ovarian RNA from postnatal day 7 onwards. The main transcripts in the testis were at all ages 7.0 kb and 2.5 kb in size. In the ovary, the main transcript was always 2.5 kb in size. In order to increase the sensitivity of mRNA detection, the FSHR gene expression was also analyzed using the reverse transcriptase-polymerase chain reaction (RT-PCR) technique with primer pairs corresponding to the near full-length FSHR mRNA or to its extracellular domain. The specificity of the PCR products was verified by Southern hybridization using a nested 32P-labeled cDNA probe. The results indicated that the expression of the extracellular domain of the FSHR was first detected on day f14.5 in the testis and on day f20.5 in the ovary. The full-length mRNA appeared in both sexes 2 days later, which is in agreement with earlier measurements of appearance of FSHR binding in the rat testis (day f17.5) and ovary (day 3 post partum). In situ hybridization using an antisense cRNA probe for FSHR demonstrated that, as early in development as specific hybridization was detected, it was confined to the Sertoli cells in the testis and to granulosa cells in the ovary. When compared with the developmental onset of the LHR gene expression (our earlier data), a major difference was observed in the ovary; the message encoding the extracellular LHR domain appeared > 10 days earlier than that corresponding to the full-length LHR message. In the case of mRNAs for the testicular LHR, and for FSHR of both sexes, the difference between the developmental appearance of the truncated and full-length RNA forms was only 2 days.
在大鼠睾丸和卵巢中，分别研究了从胎儿期第12.5天或14.5天（f）到成年期促卵泡激素（FSH）受体（R）基因表达的个体发生。在用与FSHR细胞外结构域一部分相对应的cRNA探针杂交的Northern印迹中，从胎儿期第18.5天开始检测到与睾丸RNA的特异性杂交，从出生后第7天开始检测到与卵巢RNA的特异性杂交。睾丸中的主要转录本在所有年龄段大小均为7.0 kb和2.5 kb。在卵巢中，主要转录本大小始终为2.5 kb。为了提高mRNA检测的灵敏度，还使用逆转录-聚合酶链反应（RT-PCR）技术，用与接近全长FSHR mRNA或其细胞外结构域相对应的引物对分析FSHR基因表达。PCR产物的特异性通过使用嵌套的32P标记cDNA探针的Southern杂交进行验证。结果表明，FSHR细胞外结构域的表达首先在胎儿期第14.5天在睾丸中检测到，在胎儿期第20.5天在卵巢中检测到。全长mRNA在两性中均在2天后出现，这与大鼠睾丸（胎儿期第17.5天）和卵巢（产后第3天）中FSHR结合出现的早期测量结果一致。使用针对FSHR的反义cRNA探针进行原位杂交表明，早在检测到特异性杂交时，它就局限于睾丸中的支持细胞和卵巢中的颗粒细胞。与促黄体生成素受体（LHR）基因表达的发育起始（我们早期的数据）相比，在卵巢中观察到一个主要差异；编码细胞外LHR结构域的信息比对应于全长LHR信息的信息早出现> 10天。对于睾丸LHR的mRNA以及两性的FSHR，截短和全长RNA形式的发育出现之间的差异仅为2天。
