Localization of voltage-gated K(+) channels in squid giant axons.
Localization of voltage-gated K(+) channels in squid giant axons.
复制标题
鱿鱼巨轴突中电压门控 K( ) 通道的定位。
DOI：
10.1002/1097-4695(20001115)45:3
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发表时间：
2000
期刊：
影响因子：
--
通讯作者：
Kuzirian,AM
中科院分区：
文献类型：
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作者：
Clay,JR;Kuzirian,AM
我们已经本地化的经典电压门控K+通道内鱿鱼巨轴突免疫细胞化学使用Kv 1抗体Rosenthal等。（1996年）。在轴突膜中观察到广泛分散的强烈免疫荧光斑。在轴浆中也观察到点状免疫荧光，并定位于沿神经长度方向的直径为25-50 μ m的柱（轴突直径为500 μm）。轴浆的免疫电子显微镜显示在该柱内含有直径为30-50 nm的囊泡的K+通道。这些和其他类似大小的囊泡分离轴浆使用一种新的组合高速ultracentragation和可控孔径，玻璃珠分离柱技术。约1%的所有分离的囊泡被标记的K+通道免疫金反应抗体。将分离的囊泡组分掺入人工脂质双层中，发现K+通道的电活性与拉诺等人直接从轴突膜记录的相似。（1988年）。这些含有K+通道的囊泡可能参与K+通道蛋白进入轴突膜的循环。我们还分离出了一个轴浆部分，其中含有直径为150 nm的囊泡，可以将K+通道转运回细胞体。J Neurobiol 45：172-184，2000
We have localized the classical voltage-gated K+ channel within squid giant axons by immunocytochemistry using the Kv1 antibody of Rosenthal et al.(1996). Widely dispersed patches of intense immunofluorescence were observed in the axonal membrane. Punctate immunofluorescence was also observed in the axoplasm and was localized to∼ 25–50-μm-wide column down the length of the nerve (axon diameter∼ 500 μm). Immunoelectronmicroscopy of the axoplasm revealed a K+ channel containing vesicles, 30–50 nm in diameter, within this column. These and other vesicles of similar size were isolated from axoplasm using a novel combination of high-speed ultracentrifugation and controlled-pore size, glass bead separation column techniques. Approximately 1% of all isolated vesicles were labeled by K+ channel immunogold reacted antibody. Incorporation of isolated vesicle fractions within an artificial lipid bilayer revealed K+ channel electrical activity similar to that recorded directly from the axonal membrane by Llano et al.(1988). These K+ channel–containing vesicles may be involved in cycling of K+ channel protein into the axonal membrane. We have also isolated an axoplasmic fraction containing∼ 150-nm-diameter vesicles that may transport K+ channels back to the cell body.© 2000 John Wiley & Sons, Inc. J Neurobiol 45: 172–184, 2000