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小鼠视网膜胆碱能无长突细胞中电压门控离子通道的特性分析

Characterization of voltage-gated ionic channels in cholinergic amacrine cells in the mouse retina.

作者信息

Kaneda Makoto, Ito Koichi, Morishima Yosuke, Shigematsu Yasuhide, Shimoda Yukio

机构信息

Department of Physiology, Keio University School of Medicine, Tokyo 160-8582, Japan.

出版信息

J Neurophysiol. 2007 Jun;97(6):4225-34. doi: 10.1152/jn.01022.2006. Epub 2007 Apr 11.

Abstract

Recent studies have shown that cholinergic amacrine cells possess unique membrane properties. However, voltage-gated ionic channels in cholinergic amacrine cells have not been characterized systematically. In this study, using electrophysiological and immunohistochemical techniques, we examined voltage-gated ionic channels in a transgenic mouse line the cholinergic amacrine cells of which were selectively labeled with green fluorescent protein (GFP). Voltage-gated K(+) currents contained a 4-aminopyridine-sensitive current (A current) and a tetraethylammonium-sensitive current (delayed rectifier K(+) current). Voltage-gated Ca(2+) currents contained a omega-conotoxin GVIA-sensitive component (N-type) and a omega-Aga IVA-sensitive component (P/Q-type). Tetrodotoxin-sensitive Na(+) currents and dihydropyridine-sensitive Ca(2+) currents (L-type) were not observed. Immunoreactivity for the Na channel subunit (Pan Nav), the K channel subunits (the A-current subunits [Kv. 3.3 and Kv 3.4]) and the Ca channel subunits (alpha1(A) [P/Q-type], alpha1(B) [N-type] and alpha1(C) [L-type]) was detected in the membrane fraction of the mouse retina by Western blot analysis. Immunoreactivity for the Kv. 3.3, Kv 3.4, alpha1(A) [P/Q-type], and alpha1(B) [N-type] was colocalized with the GFP signals. Immunoreactivity for alpha1(C) [L-type] was not colocalized with the GFP signals. Immunoreactivity for Pan Nav did not exist on the membrane surface of the GFP-positive cells. Our findings indicate that signal propagation in cholinergic amacrine cells is mediated by a combination of two types of voltage-gated K(+) currents (the A current and the delayed rectifier K(+) current) and two types of voltage-gated Ca(2+) currents (the P/Q-type and the N-type) in the mouse retina.

摘要

最近的研究表明,胆碱能无长突细胞具有独特的膜特性。然而,胆碱能无长突细胞中的电压门控离子通道尚未得到系统的表征。在本研究中,我们使用电生理和免疫组织化学技术,在一个转基因小鼠品系中检测电压门控离子通道,该品系的胆碱能无长突细胞被绿色荧光蛋白(GFP)选择性标记。电压门控钾电流包含一个4-氨基吡啶敏感电流(A电流)和一个四乙铵敏感电流(延迟整流钾电流)。电压门控钙电流包含一个ω-芋螺毒素GVIA敏感成分(N型)和一个ω-银环蛇毒素IVA敏感成分(P/Q型)。未观察到河豚毒素敏感的钠电流和二氢吡啶敏感的钙电流(L型)。通过蛋白质免疫印迹分析在小鼠视网膜的膜部分检测到钠通道亚基(泛钠通道)、钾通道亚基(A电流亚基[Kv. 3.3和Kv 3.4])和钙通道亚基(α1(A)[P/Q型]、α1(B)[N型]和α1(C)[L型])的免疫反应性。Kv. 3.3、Kv 3.4、α1(A)[P/Q型]和α1(B)[N型]的免疫反应性与GFP信号共定位。α1(C)[L型]的免疫反应性与GFP信号未共定位。GFP阳性细胞的膜表面不存在泛钠通道的免疫反应性。我们的研究结果表明,在小鼠视网膜中,胆碱能无长突细胞中的信号传播是由两种类型的电压门控钾电流(A电流和延迟整流钾电流)和两种类型的电压门控钙电流(P/Q型和N型)共同介导的。

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