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永生化下丘脑GT1-7神经元表达功能性A型γ-氨基丁酸受体。

Immortalized hypothalamic GT1-7 neurons express functional gamma-aminobutyric acid type A receptors.

作者信息

Hales T G, Kim H, Longoni B, Olsen R W, Tobin A J

机构信息

Department of Anesthesiology, School of Medicine, University of California, Los Angeles 90024.

出版信息

Mol Pharmacol. 1992 Aug;42(2):197-202.

PMID:1325030
Abstract

Neuronal cell lines provide a source of pure populations of neurons and allow the properties of many neurotransmitter receptors to be studied. However, none of these cells have been reported to express functional gamma-aminobutyric acid (GABA)A receptors. Indeed, there have been no reports of cell lines expressing functional amino acid receptors. Using biochemical and electrophysiological techniques, we have identified a neuronal cell line expressing functional GABAA receptors. Membranes from immortalized hypothalamic (GT1-7) neurons bound [3H]muscimol but not [3H]flunitrazepam. GABA-activated chloride currents, recorded from GT1-7 cells, were blocked by bicuculline and Zn2+ but were insensitive to diazepam. These results suggest that GABAA receptors on GT1-7 cells lack gamma subunits. The neurosteroid 5 alpha-pregnan-3 alpha-ol-20-one and pentobarbital both modulated GABAA receptors in these cells. Polymerase chain reaction analysis of the cells revealed the presence of mRNAs encoding alpha 1, beta 1, and beta 3 polypeptides. GT1-7 cells provide a useful model system for studying the regulation of GABAA receptor polypeptide expression.

摘要

神经元细胞系为纯神经元群体提供了来源,并使得许多神经递质受体的特性得以研究。然而,尚无报道称这些细胞中有任何一种表达功能性γ-氨基丁酸(GABA)A受体。事实上,也没有关于表达功能性氨基酸受体的细胞系的报道。我们运用生化和电生理技术,鉴定出了一种表达功能性GABAA受体的神经元细胞系。永生化下丘脑(GT1-7)神经元的细胞膜能结合[3H]蝇蕈醇,但不能结合[3H]氟硝西泮。从GT1-7细胞记录到的GABA激活的氯离子电流被荷包牡丹碱和Zn2+阻断,但对地西泮不敏感。这些结果表明,GT1-7细胞上的GABAA受体缺乏γ亚基。神经甾体5α-孕烷-3α-醇-20-酮和戊巴比妥均可调节这些细胞中的GABAA受体。对这些细胞进行聚合酶链反应分析,结果显示存在编码α1、β1和β3多肽的mRNA。GT1-7细胞为研究GABAA受体多肽表达的调控提供了一个有用的模型系统。

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