Cellular localization of voltage-gated calcium channels and synaptic vesicle-associated proteins in the guinea pig cochlea
Cellular localization of voltage-gated calcium channels and synaptic vesicle-associated proteins in the guinea pig cochlea
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DOI：
10.1385/jmn:27:2:225
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发表时间：
2005-01-01
影响因子：
3.1
通讯作者：
Yates, GK
中科院分区：
文献类型：
--
作者：
Layton, MG;Robertson, D;Yates, GK
应用免疫细胞化学和激光共聚焦显微镜研究了豚鼠耳蜗内电压门控钙通道（VGCC）和突触囊泡相关蛋白SV 2、突触蛋白1和囊泡相关膜蛋白（VAMP）的细胞定位。在豚鼠中，通过Western印迹和小脑的免疫标记来测定抗体对L型，α（1C）[Ca（nu）1.2]和α（1D）[Ca（v）1.3]; P/Q型，α（1A）[Ca（nu）2.1];和R型，α（1 E）[Ca（nu）2.3]高压激活钙通道的α（1）亚基的反应性。在耳蜗的感觉内毛细胞的器官的Corti表现出强烈的细胞内染色，主要定位于其基底外侧极，与抗体直接针对的oc，c亚基。一些α（1C）标记也观察到在内柱细胞，在螺旋神经节的传入神经元的细胞体，并在螺旋韧带的下部区域。支持柱细胞的α（1D）免疫反应性很强，但没有α（1D），内毛细胞的标记。α（1A）亚基在所有三排外毛细胞中均呈胞质分布。α（1 E）标记定位于外毛细胞，主要位于表皮下板区域，也定位于这些毛细胞下方的神经纤维束。强烈的免疫反应性一致地看到与针对SV 2和突触蛋白I的抗体在神经元结构周围的基底外侧表面的内毛细胞和外毛细胞，但不存在从感觉细胞本身。VAMP标记被发现在整个内毛细胞的细胞质和毛细胞下方的神经元结构。这些结果揭示了豚鼠耳蜗的感觉和非感觉元件中VGCC类型的差异分布，内毛细胞表达α L-1C型通道和VAMP，但不表达突触蛋白I或SV 2。
The cellular localization of voltage-gated calcium channels (VGCCs) and synaptic vesicle-associated proteins, SV2, synapsin 1, and vesicle-associated membrane protein (VAMP) (synaptobrevin), was investigated in the guinea pig cochlea using immunocytochernistry and confocal laser scanning microscopy. Reactivity, in guinea pig, of antibodies to the alpha(1) subunits of L-type, alpha(1C) [Ca(nu)1.2] and alpha(1D) [Ca(v)1.3]; P/Q-type, alpha(1A)[Ca(nu)2.1]; and R-type, alpha(1E) [Ca(nu)2.3] high voltage-activated calcium channels, was determined by Western blotting and immunolabeling of cerebellum. In the cochlea the sensory inner hair cells of the organ of Corti displayed strong intracellular staining, predominantly localized to their basolateral poles, with an antibody directed against the oc,c subunit. Some alpha(1C) labeling was also observed in the inner pillar cells, in cell bodies of afferent neurons in the spiral ganglion, and in the inferior region of the spiral ligament. The supporting pillar cells were strongly immunoreactive throughout for alpha(1D), but no alpha(1D), labeling of the inner hair cells was seen. The alpha(1A), subunit showed a cytoplasmic distribution in all three rows of outer hair cells. alpha(1E) labeling localized to the outer hair cells, predominantly in the subcuticular plate region, and also to nerve fiber bundles beneath these hair cells. Strong immunoreactivity was consistently seen with antibodies directed against SV2 and synapsin I in neuronal structures surrounding the basolateral surfaces of both the inner and outer hair cells but was absent from the sensory cells themselves. VAMP labeling was found throughout the cytoplasm of the inner hair cells and in neuronal structures beneath the hair cells. These results reveal a differential distribution of VGCC-types in the sensory and nonsensory elements of the guinea pig cochlea, with the inner hair cells expressing alpha L-1C-type channels and VAMP but not synapsin I or SV2.