Rossi P, Dolci S, Albanesi C, Grimaldi P, Ricca R, Geremia R
Department of Public Health and Cell Biology, University of Rome Tor Vergata, Italy.
Dev Biol. 1993 Jan;155(1):68-74. doi: 10.1006/dbio.1993.1007.
Follicle-stimulating hormone (FSH) and its intracellular mediator, cAMP, increase the mRNA levels for the Steel factor (SLF, the c-kit ligand) in cultured primary mouse Sertoli cells. The inductive effect of cAMP is more evident in cultures from 13-day-old animals than in cultures from 18-day-old animals. Analysis through the polymerase chain reaction (PCR) indicates that (Bu)2cAMP or FSH treatment increases the levels of the mRNAs for both the potentially soluble form and the transmembrane form of SLF in cultured Sertoli cells. The ratio between mRNAs encoding the potentially soluble form and the transmembrane form of SLF increases during postnatal testis development, and it is higher in cultured Sertoli cells with respect to total testis, suggesting that, under the in vitro conditions, SLF could be produced by Sertoli cells mainly as a soluble factor. Soluble recombinant SLF stimulates, in a dose-dependent fashion, thymidine incorporation in cultures of isolated germ cell populations enriched in the mitotic stages (spermatogonia), independently of the presence of serum, whereas cAMP analogs have no effect. Autoradiographic analysis shows that SLF selectively stimulates DNA synthesis in type A spermatogonia.
促卵泡激素(FSH)及其细胞内介质环磷酸腺苷(cAMP)可提高原代培养的小鼠支持细胞中干细胞因子(SLF,即c-kit配体)的mRNA水平。cAMP的诱导作用在13日龄动物的培养物中比在18日龄动物的培养物中更明显。通过聚合酶链反应(PCR)分析表明,(Bu)2cAMP或FSH处理可提高培养的支持细胞中潜在可溶性形式和跨膜形式的SLF的mRNA水平。编码潜在可溶性形式和跨膜形式的SLF的mRNA之间的比率在出生后睾丸发育过程中增加,并且在培养的支持细胞中相对于整个睾丸更高,这表明在体外条件下,SLF可能主要作为一种可溶性因子由支持细胞产生。可溶性重组SLF以剂量依赖的方式刺激富含有丝分裂阶段(精原细胞)的分离生殖细胞群体培养物中的胸腺嘧啶掺入,与血清的存在无关,而cAMP类似物则没有作用。放射自显影分析表明,SLF选择性地刺激A型精原细胞中的DNA合成。