TACHYKININ (SUBSTANCE-P) GENE-EXPRESSION IN LEYDIG-CELLS OF THE HUMAN AND MOUSE TESTIS
TACHYKININ (SUBSTANCE-P) GENE-EXPRESSION IN LEYDIG-CELLS OF THE HUMAN AND MOUSE TESTIS
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DOI：
10.1210/endo-128-5-2441
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发表时间：
1991-05-01
期刊：
影响因子：
4.8
通讯作者：
IVELL, R
中科院分区：
文献类型：
--
作者：
CHIWAKATA, C;BRACKMANN, B;IVELL, R
P物质免疫反应在间质细胞，尤其是人睾丸中可检测到，在小鼠间质细胞中检测到较少，但在大鼠中检测不到。用改进的聚合酶链式反应(PCR)方法在人、小鼠和牛的睾丸提取液中检测到速激肽前A(物质-P)mRNA，而在大鼠、猪的睾丸、作为阴性对照的牛的甲状腺和黄体中检测不到。这项检测能够区分该基因的α、β和伽马转录本，并表明只有贝塔和伽马转录本存在于睾丸中。对牛下丘脑、小鼠脑和人睾丸的PCR产物进行测序分析，证实了这些转录本的结构，这些转录本编码P物质和神经激肽A(K物质)神经肽激素。使用这种方法的一种变种，可以在500个新鲜制备的纯化的小鼠间质细胞中鉴定速激肽的转录本。在平行研究中，聚合酶链式反应分析也能够证实人类睾丸中存在P物质和神经激动素A受体的mRNA。因此，速激肽物质-P和神经激动素-A现在必须被添加到调节睾丸内功能的潜在旁分泌物质的列表中。
Specific substance-P immunoreactivity can be detected in the Leydig cells, particularly of human testes, and to a lesser degree in mouse Leydig cells, but not in the rat. Using a modified polymerase chain reaction (PCR) assay, preprotachykinin-A (substance-P) mRNA could be detected in extracts of human, mouse, and bovine testes, but not in rat or boar testes or in bovine thyroid or corpus luteum used as negative controls. This assay is able to discriminate among the alpha, beta, and gamma transcripts of the gene and shows that only the beta and gamma transcripts are present in the testes. Sequencing analysis of the PCR products from bovine hypothalamus, mouse brain, and human testis confirmed the structure of these transcripts, which encode both substance-P and neurokinin-A (substance-K) neuropeptide hormones. Using a variant of this assay it was possible to identify tachykinin transcripts in as few as 500 freshly prepared purified mouse Leydig cells. In parallel studies PCR analysis was also able to confirm the presence of mRNA for both substance-P and neurokinin-A receptors in human testes. Thus, the tachykinins substance-P and neurokinin-A must now be added to the list of potentially paracrine substances regulating intratesticular function.