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大鼠围产期脑、卵巢及一种睾丸间质细胞瘤细胞系中芳香化酶细胞色素P-450 mRNA的特性分析:脑特异性芳香化酶转录本存在的证据

Characterization of aromatase cytochrome P-450 mRNA in rat perinatal brain, ovary and a Leydig tumor cell line: evidence for the existence of brain specific aromatase transcripts.

作者信息

Lephart E D, Herbst M A, McPhaul M J

机构信息

Department of Internal Medicine, The University of Texas Southwestern Medical Center at Dallas, 5323 Harry Hines Boulevard, 75235-8857, Dallas, Texas, USA.

出版信息

Endocrine. 1995 Jan;3(1):25-31. doi: 10.1007/BF02917445.

Abstract

The conversion of androgens to estrogens is catalyzed by the aromatase cytochrome P-450 (P-450(AROM)) in a veriety of tissues and cell types in vertebrates. The manner in which aromatase activity is regulated appears to be quite different, even between different tissues of a single species. In the current study, we have determined the sequence of the 5' end of the aromatase mRNA in a rat Leydig tumor cell line, R2C, and in three rat tissue [adult ovary, perinatal amygdala (AmY), and medial basal hypothalamus-preoptic area (MBH-POA)], each of which shows different patterns of aromatase expression. S(1) nuclease protection and primer-extension analyses establish that the site of transcription initiation of the aromatase mRNA present in rat ovarh and the rat Leydig tumor cell line R2C is located approximately 97 nucleotides upstream from the initiator methionine. By contrast, although aromatase mRNA was detected in S(1) nuclease protection experiments using a probe derived from the aromatase open-reading frame, transcripts initiating at this site were absent from RNA samples prepared from perinatal rat AMY and MBH-POA tissue. S(1) mapping and sequencing of the 5' end of AMY and MBH-POA aromatase cDNAs indicate that the aromatase mRNA transcripts present in these rat neural tissues from perinatal animals contain a distinctive 5' terminus and are derived from a different promoter.

摘要

雄激素向雌激素的转化是由芳香化酶细胞色素P - 450(P - 450(AROM))在脊椎动物的多种组织和细胞类型中催化完成的。芳香化酶活性的调节方式似乎差异很大,即使在单一物种的不同组织之间也是如此。在本研究中,我们确定了大鼠睾丸间质细胞瘤细胞系R2C以及三种大鼠组织[成年卵巢、围产期杏仁核(AmY)和内侧基底下丘脑 - 视前区(MBH - POA)]中芳香化酶mRNA 5'端的序列,每种组织都呈现出不同的芳香化酶表达模式。S(1)核酸酶保护分析和引物延伸分析表明,大鼠卵巢和大鼠睾丸间质细胞瘤细胞系R2C中存在的芳香化酶mRNA转录起始位点位于起始甲硫氨酸上游约97个核苷酸处。相比之下,尽管在使用源自芳香化酶开放阅读框的探针进行的S(1)核酸酶保护实验中检测到了芳香化酶mRNA,但从围产期大鼠AmY和MBH - POA组织制备的RNA样本中却没有从该位点起始的转录本。对AmY和MBH - POA芳香化酶cDNA 5'端的S(1)定位和测序表明,围产期动物的这些大鼠神经组织中存在的芳香化酶mRNA转录本含有一个独特的5'末端,并且来自不同的启动子。

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