Terasaki M, Schmidek A, Galbraith J A, Gallant P E, Reese T S
Laboratory of Neurobiology, National Institute of Neurological Disorders and Stroke, National Institutes of Health, Bethesda, MD 20892-4062, USA.
Proc Natl Acad Sci U S A. 1995 Dec 5;92(25):11500-3. doi: 10.1073/pnas.92.25.11500.
In order to explore how cytoskeletal proteins are moved by axonal transport, we injected fluorescent microtubules and actin filaments as well as exogenous particulates into squid giant axons and observed their movements by confocal microscopy. The squid giant axon is large enough to allow even cytoskeletal assemblies to be injected without damaging the axon or its transport mechanisms. Negatively charged, 10- to 500-nm beads and large dextrans moved down the axon, whereas small (70 kDa) dextrans diffused in all directions and 1000-nm beads did not move. Only particles with negative charge were transported. Microtubules and actin filaments, which have net negative charges, made saltatory movements down the axon, resulting in a net rate approximating that previously shown for slow transport of cytoskeletal elements. The present observations suggest that particle size and charge determine which materials are transported down the axon.
为了探究细胞骨架蛋白如何通过轴突运输移动,我们将荧光微管、肌动蛋白丝以及外源性颗粒注射到枪乌贼的巨大轴突中,并通过共聚焦显微镜观察它们的移动。枪乌贼的巨大轴突足够大,甚至可以注射细胞骨架组件而不损伤轴突或其运输机制。带负电荷的10至500纳米珠子和大葡聚糖沿轴突向下移动,而小(70 kDa)葡聚糖向各个方向扩散,1000纳米珠子则不移动。只有带负电荷的颗粒被运输。带有净负电荷的微管和肌动蛋白丝沿轴突进行跳跃式移动,其净速率接近先前显示的细胞骨架元件缓慢运输的速率。目前的观察结果表明,颗粒大小和电荷决定了哪些物质沿轴突运输。