Evans L L, Bridgman P C
Washington University School of Medicine, Department of Anatomy and Neurobiology, St. Louis, MO 63110, USA.
Proc Natl Acad Sci U S A. 1995 Nov 21;92(24):10954-8. doi: 10.1073/pnas.92.24.10954.
Organelle movement along actin filaments has been demonstrated in dissociated squid axoplasm [Kurznetsov, S. A., Langford, G.M. & Weiss, D. G. (1992) Nature (London) 356, 722-725 and Bearer, E.L., DeGiorgis, J.A., Bodner, R.A., Kao, A.W. & Reese, T.S. (1993) Proc. Natl. Acad. Sci. USA 90, 11252-11256] but has not been shown to occur in intact neurons. Here we demonstrate that intracellular transport occurs along actin filament bundles in intact neuronal growth cones. We used video-enhanced differential interference contrast microscopy to observe intracellular transport in superior cervical ganglion neurons cultured under conditions that enhance the visibility of actin bundles within growth cone lamellipodia. Intracellular particles, ranging in size from < 0.5-1.5 microns, moved along linear structures (termed transport bundles) at an average maximum rate of 0.48 micron/sec. After particle movement had been viewed, cultures were preserved by rapid perfusion with chemical fixative. To determine whether particle transport occurred along actin, we then used fluorescence microscopy to correlate this movement with actin and microtubule distributions in the same growth cones. The observed transport bundles colocalized with actin but not with microtubules. The rates of particle movement and the association of moving particles with actin filament bundles suggest that myosins may participate in the transport of organelles (or other materials) in intact neurons.
在分离的枪乌贼轴质中已证实细胞器沿肌动蛋白丝移动[库尔兹涅佐夫,S. A.,兰福德,G. M. & 韦斯,D. G.(1992年)《自然》(伦敦)356, 722 - 725;以及贝勒,E. L.,德乔吉斯,J. A.,博德纳,R. A.,考,A. W. & 里斯,T. S.(1993年)《美国国家科学院院刊》90, 11252 - 11256],但尚未证明其在完整神经元中发生。在此我们证明,在完整的神经元生长锥中,细胞内运输沿肌动蛋白丝束发生。我们使用视频增强微分干涉对比显微镜来观察在增强生长锥片足内肌动蛋白束可见性的条件下培养的颈上神经节神经元中的细胞内运输。大小从<0.5 - 1.5微米不等的细胞内颗粒沿着线性结构(称为运输束)移动,平均最大速率为0.48微米/秒。在观察到颗粒移动后,通过用化学固定剂快速灌注来保存培养物。为了确定颗粒运输是否沿肌动蛋白发生,然后我们使用荧光显微镜将这种移动与同一生长锥中的肌动蛋白和微管分布相关联。观察到的运输束与肌动蛋白共定位,但与微管不共定位。颗粒移动的速率以及移动颗粒与肌动蛋白丝束的关联表明,肌球蛋白可能参与完整神经元中细胞器(或其他物质)的运输。