ORGANELLE TRANSPORT OF ION CHANNELS IN EXCITABLE CELLS

可兴奋细胞中离子通道的细胞器运输

• 批准号: 2579548
• 负责人: J R CLAY
• 金额: --
• 依托单位: NATIONAL INSTITUTE OF NEUROLOGICAL DISORDERS AND STROKE
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/2579548
• 关键词: alternatives to animals in research; axoplasm; cell motility; confocal scanning microscopy; fluorescence microscopy; fluorescent dye /probe; interference microscopy; ion transport; membrane channels; microtubules; neurofilament; neuronal transport; organelles; squid

This project concerns the transport of ion channels in nerve axoplasm by intracellular organelles. Previous work in this project has demonstrated distinct populations of organelles isolated from the axoplasm of squid giant axons; a putative anterograde organelle population having diameters in the 40-60 nm range, and a putative retrograde population having diameters in the 100-150 nm range. During the past year various microscopic techniques have been applied in an attempt to visualize the organelles in various different settings. For example, laser confocal microscopy has recently been used to observe Brownian motion of organelles which had been labeled with the fluorescent marker, Texas Red. These results are being used to determine the relative yield of organelles in the various fractions from the column, and also to provide some information on organelle size from the Brownian motion. The Texas Red treated preparations are also being used in an attempt to visualize motility of exogenous organelles along microtubules and perhaps also actin filaments in an in vitro motility assay with the differential interference contrast microscopy technique, and they are also being pressure injected into squid axons to determine the targeting of organelles to the axonal membrane with a fluorescence microscope.

本项目涉及神经轴浆中离子通道的转运， 胞内细胞器 该项目的前期工作表明， 从鱿鱼轴质中分离的不同细胞器群体 巨大的轴突;一个假定的顺行细胞器群体， 在40-60纳米范围内，和一个假定的逆行人口， 直径在100-150 nm范围内。 在过去的一年里， 显微技术已经被应用于试图可视化 各种不同的环境中的细胞器。 例如，激光共聚焦 显微镜最近被用来观察布朗运动， 这些细胞器已经用荧光标记物德克萨斯红标记。 这些结果被用来确定相对产量 细胞器中的各种馏分从柱，也提供 从布朗运动中得到一些关于细胞器大小的信息。 德州 红色处理的制剂也被用于试图可视化 外源性细胞器沿着微管运动， 肌动蛋白丝在体外运动试验中的差异 干涉对比显微镜技术，他们也正在 压力注入鱿鱼轴突，以确定目标， 用荧光显微镜观察轴突膜上的细胞器。