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[耳蜗组织中γ-氨基丁酸A受体mRNA的检测。原位杂交研究]

[Detection of GABA(A) receptor mRNA in cochlear tissue. An in situ hybridization study].

作者信息

Kempf H G, Brändle T U, Wisden W, Zenner H P, Marx A

机构信息

HNO-Klinik, Medizinischen Hochschule Hannover.

出版信息

HNO. 1995 Jan;43(1):12-8.

PMID:7890545
Abstract

Gamma-aminobutyric acid (GABA) and the GABAergic system play an important role in the efferent modulation of cochlear function. We examined surface preparations of guinea pig and mouse cochleae by in situ hybridization using radioactive labelled oligonucleotides for several subunits of the GABAA receptor. Frozen sections of rat and guinea pig brain (cortex, hippocampus, cerebellum) served as controls. In the mouse cochlea the mRNA of the alpha-1 and alpha-5, beta-1 and gamma-1 subunit were detected, while in guinea pig cochlea mRNA of the alpha-1, alpha-4, alpha-5, and gamma-1 subunit of the GABAA receptor were found. Positive signals were located in the regions of the outer hair cells and had a weaker intensity in the inner hair cells. In the brain sections the several subunits were detected in a variable distribution in the cerebellum, hippocampus and cortical regions. Rat specimens exhibited stronger signals than guinea pig brain sections. These investigations have extended previous results of immunocytochemical experiments from our laboratory demonstrating mRNA sequences of GABAA receptor subunits in the mammalian inner ear. Detection of these nucleotide sequences using surface preparations of the cochlea on a molecular level by in situ hybridization supports the importance of GABA as a cochlear neurotransmitter. Furthermore, it can be concluded that the mammalian cochlea is able to express a GABA-dependent neurotransmission system.

摘要

γ-氨基丁酸(GABA)及其能系统在耳蜗功能的传出调制中发挥重要作用。我们使用放射性标记的寡核苷酸对GABAA受体的几个亚基进行原位杂交,检查了豚鼠和小鼠耳蜗的表面标本。大鼠和豚鼠脑(皮质、海马、小脑)的冰冻切片用作对照。在小鼠耳蜗中检测到α-1、α-5、β-1和γ-1亚基的mRNA,而在豚鼠耳蜗中发现了GABAA受体的α-1、α-4、α-5和γ-1亚基的mRNA。阳性信号位于外毛细胞区域,在内毛细胞中强度较弱。在脑切片中,在小脑、海马和皮质区域检测到几个亚基的分布各不相同。大鼠标本的信号比豚鼠脑切片更强。这些研究扩展了我们实验室先前免疫细胞化学实验的结果,这些结果证明了哺乳动物内耳中GABAA受体亚基的mRNA序列。通过原位杂交在分子水平上使用耳蜗表面标本检测这些核苷酸序列,支持了GABA作为耳蜗神经递质的重要性。此外,可以得出结论,哺乳动物耳蜗能够表达GABA依赖性神经传递系统。

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