Chen J K, Heckert L L
Department of Molecular and Integrative Physiology, The University of Kansas Medical Center, Kansas City, Kansas 66160, USA.
Endocrinology. 2001 Mar;142(3):1167-78. doi: 10.1210/endo.142.3.8021.
Dmrt1 is a recently described gene that is expressed exclusively in the testis and is required for postnatal testis differentiation. Here we describe the expression of Dmrt1 in postnatal rat testis and Sertoli cells. RNase protection analysis was used to examine Dmrt1 messenger RNA (mRNA) levels in intact testis during postnatal development and in primary cultures of Sertoli cells under various culture conditions. We show that Dmrt1 mRNA levels rise significantly beginning approximately 10 days after birth and remain elevated until after the third postnatal week. Thereafter, mRNA levels drop coincident with the proliferation of germ cells in the testis. In freshly isolated Sertoli cells, Dmrt1 mRNA levels were robust but decreased significantly when the cells were placed in culture for 24 h. Treatment of Sertoli cells with either FSH or 8-bromo-cAMP resulted in a significant rise in Dmrt1 mRNA levels. This cAMP response was sensitive to treatment with the transcriptional inhibitor actinomycin D but not to the translational inhibitor cycloheximide. The cAMP-dependent rise in Dmrt1 mRNA also required activation of protein kinase A, as mRNA induction was sensitive to the inhibitor H89. Studies also show that Dmrt1 expression was inhibited by phorbol esters (PMA) but only modestly effected by serum.
Dmrt1是一个最近被描述的基因,它仅在睾丸中表达,是出生后睾丸分化所必需的。在这里,我们描述了Dmrt1在出生后大鼠睾丸和支持细胞中的表达。采用核糖核酸酶保护分析法检测出生后发育过程中完整睾丸以及在各种培养条件下支持细胞原代培养物中Dmrt1信使核糖核酸(mRNA)水平。我们发现,Dmrt1 mRNA水平在出生后约10天开始显著升高,并一直保持升高,直到出生后第三周之后。此后,mRNA水平随着睾丸中生精细胞的增殖而下降。在新鲜分离的支持细胞中,Dmrt1 mRNA水平很高,但当细胞培养24小时后显著下降。用促卵泡激素(FSH)或8-溴环磷腺苷(8-bromo-cAMP)处理支持细胞会导致Dmrt1 mRNA水平显著升高。这种环磷腺苷反应对转录抑制剂放线菌素D的处理敏感,但对翻译抑制剂环己酰亚胺不敏感。Dmrt1 mRNA的环磷腺苷依赖性升高也需要蛋白激酶A的激活,因为mRNA的诱导对抑制剂H89敏感。研究还表明,佛波酯(PMA)可抑制Dmrt1的表达,但血清对其影响较小。