IMMUNOCYTOCHEMICAL LOCALIZATION OF SODIUM-CHANNELS IN AN INSECT CENTRAL-NERVOUS-SYSTEM USING A SITE-DIRECTED ANTIBODY
IMMUNOCYTOCHEMICAL LOCALIZATION OF SODIUM-CHANNELS IN AN INSECT CENTRAL-NERVOUS-SYSTEM USING A SITE-DIRECTED ANTIBODY
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DOI：
10.1002/neu.480240707
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发表时间：
1993-07-01
期刊：
影响因子：
--
通讯作者：
MURPHY, RA
中科院分区：
文献类型：
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作者：
FRENCH, AS;SANDERS, EJ;MURPHY, RA
针对通道蛋白和特定肽序列的抗体以前曾被用来定位大鼠大脑中的电压激活钠通道。在这里，我们描述了首次使用定位抗体定位昆虫神经系统中的钠通道。用高度保守的SP19序列的抗体对蟑螂的中胸神经节进行染色。在蜡片上用亲和素/生物素方法进行光镜观察，在免疫金标记的薄片上用透射电子显微镜观察抗体标记。昆虫的中央神经节含有明显分离的细胞体、突触神经束、轴索束和神经。光镜下的抗体染色仅限于神经元，在神经节和神经的轴突中染色强烈。在许多神经元胞体的胞浆中也有很强的染色，但在胞核中没有。神经纤维区的标记相对较浅。这些发现可以与已知的神经节电生理学相关联。电子显微镜在轴突周围区域检测到钠通道，可能包括轴突膜和包裹神经胶质细胞膜。轴突线粒体也被大量标记，表明这些细胞器具有钠通道运输功能。
Antibodies to channel proteins and specific peptide sequences have been previously used to localize voltage-activated sodium channels in the rat brain. Here we describe the first localization of sodium channels in an insect nervous system using a site-directed antibody. The mesothoracic ganglion of the cockroach was stained with an antibody to the highly conserved SP19 sequence. Antibody labelling was visualized by light microscopy using the avidin/biotin method on wax sections, and transmission electron microscopy of immunogold-labelled thin sections. Central ganglia of insects contain clearly separated regions of cell bodies, synaptic neuropil, axon tracts, and nerves. Antibody staining by light microscopy was limited to neurons, and was intense in axons throughout the ganglion and nerves. Staining was also strong in the cytoplasm, but not the nuclei, of many neuronal cell bodies. Neuropil regions were relatively lightly labelled. These findings can be correlated with the known electrophysiology of the ganglion. Electron microscopy detected sodium channels in areas surrounding axons, probably including axon membranes and enveloping glial cell membranes. Axonal mitochondria were also heavily labelled, suggesting a sodium channel transport function for these organelles.