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中脑多巴胺能神经元中γ-氨基丁酸A型(GABAA)受体亚基变体的鉴定

Identification of GABAA receptor subunit variants in midbrain dopaminergic neurons.

作者信息

Okada Hideki, Matsushita Natsuki, Kobayashi Kenta, Kobayashi Kazuto

机构信息

Department of Molecular Genetics, Institute of Biomedical Sciences, Fukushima Medical University School of Medicine, Fukushima, Japan.

出版信息

J Neurochem. 2004 Apr;89(1):7-14. doi: 10.1111/j.1471-4159.2004.02271.x.

Abstract

Modulation of the activity of dopamine (DA)-producing neurons by GABA plays an important role in the control of DA-mediated brain functions. Ionotropic GABA(A) receptors exist as heteropentametric structures assembling different subunits composed of various subtypes. However, the expression pattern of these subunits in DA neurons in the ventral midbrain has not been fully defined. In the present study, we investigated the subunit composition of GABA(A) receptors in DA neurons in the substantia nigra pars compacta (SNc) and the ventral tegmental area (VTA). We isolated DA neurons from the ventral midbrain of transgenic mice that express green fluorescent protein under the control of the tyrosine hydroxylase (TH) gene promoter and analyzed expression of various GABA(A) receptor subunits in single cells by using the reverse transcription-polymerase chain reaction. This analysis showed the presence of the transcripts encoding alpha2, alpha3, alpha4, beta1, beta3 and gamma2 subunits in the isolated DA neurons. Double fluorescence in situ hybridization with probes for TH and GABA(A) receptor subunit mRNAs revealed the expression of these six subunits in the majority of DA neurons in the SNc and the VTA.

摘要

γ-氨基丁酸(GABA)对多巴胺(DA)能神经元活动的调节在DA介导的脑功能控制中起着重要作用。离子型GABA(A)受体以异五聚体结构存在,由不同亚型组成不同亚基。然而,这些亚基在腹侧中脑DA神经元中的表达模式尚未完全明确。在本研究中,我们调查了黑质致密部(SNc)和腹侧被盖区(VTA)中DA神经元GABA(A)受体的亚基组成。我们从在酪氨酸羟化酶(TH)基因启动子控制下表达绿色荧光蛋白的转基因小鼠腹侧中脑分离出DA神经元,并通过逆转录-聚合酶链反应分析单细胞中各种GABA(A)受体亚基的表达。该分析显示,在分离出的DA神经元中存在编码α2、α3、α4、β1、β3和γ2亚基的转录本。用TH和GABA(A)受体亚基mRNA探针进行的双荧光原位杂交显示,这六个亚基在SNc和VTA的大多数DA神经元中表达。

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