Nitsche E M, Moquin A, Adams P S, Guenette R S, Lakins J N, Sinnecker G H, Kruse K, Tenniswood M P
W.A. Jones Cell Science Center, Lake Placid, New York, USA.
Am J Med Genet. 1996 May 3;63(1):231-8. doi: 10.1002/(SICI)1096-8628(19960503)63:1<231::AID-AJMG40>3.0.CO;2-M.
Male sexual differentiation is a process that involves androgen action via the androgen receptor. Defects in the androgen receptor, many resulting from point mutations in the androgen receptor gene, lead to varying degrees of impaired masculinization in chromosomally male individuals. To date no specific androgen regulated morphogens involved in this process have been identified and no marker genes are known that would help to predict further virilization in infants with partial androgen insensitivity. In the present study we first show data on androgen regulated gene expression investigated by differential display reverse transcription PCR (dd RT PCR) on total RNA from human neonatal genital skin fibroblasts cultured in the presence or absence of 100 nM testosterone. Using three different primer combinations, 54 cDNAs appeared to be regulated by androgens. Most of these sequences show the characteristics of expressed mRNAs but showed no homology to sequences in the database. However 15 clones with significant homology to previously cloned sequences were identified. Seven cDNAs appear to be induced by androgen withdrawal. Of these, five are similar to ETS (expression tagged sequences) from unknown genes; the other two show significant homology to the cDNAs of ubiquitin and human guanylate binding protein 2 (GBP-2). In addition, we have identified 8 cDNA clones which show homologies to other sequences in the database and appear to be upregulated in the presence of testosterone. Four of these clones again are similar to ETS from unknown genes. Three differential expressed sequences that appear to be upregulated in the presence of testosterone show significant homology to the cDNAs of L-plastin and one to the cDNA of testican. This latter gene codes for a proteoglycan involved in cell social behavior and therefore of special interest in this context. The results of this study are of interest in further investigation of normal and disturbed androgen-dependent gene expression.
男性性分化是一个涉及雄激素通过雄激素受体发挥作用的过程。雄激素受体的缺陷，其中许多是由雄激素受体基因的点突变导致的，会使染色体为男性的个体出现不同程度的男性化受损。迄今为止，尚未鉴定出参与这一过程的特定雄激素调节形态发生素，也没有已知的标记基因可用于预测部分雄激素不敏感婴儿的进一步男性化情况。在本研究中，我们首先展示了通过差异显示逆转录聚合酶链反应（dd RT PCR）对在有或无100 nM睾酮的情况下培养的人类新生儿生殖器皮肤成纤维细胞的总RNA进行研究得到的雄激素调节基因表达数据。使用三种不同的引物组合，54个cDNA似乎受雄激素调节。这些序列中的大多数显示出表达的mRNA的特征，但与数据库中的序列没有同源性。然而，鉴定出了15个与先前克隆序列具有显著同源性的克隆。七个cDNA似乎是由雄激素撤除诱导的。其中，五个与未知基因的ETS（表达标签序列）相似；另外两个与泛素和人鸟苷酸结合蛋白2（GBP - 2）的cDNA具有显著同源性。此外，我们鉴定出8个cDNA克隆，它们与数据库中的其他序列具有同源性，并且在睾酮存在的情况下似乎上调。其中四个克隆再次与未知基因的ETS相似。三个在睾酮存在的情况下似乎上调的差异表达序列与L - 肌动蛋白的cDNA具有显著同源性，另一个与睾丸蛋白聚糖的cDNA具有同源性。后一个基因编码一种参与细胞社交行为的蛋白聚糖，因此在这种情况下特别受关注。本研究结果对于进一步研究正常和紊乱的雄激素依赖性基因表达具有重要意义。
