Glass J D, Griffin J W
Department of Neurology, Johns Hopkins University School of Medicine, Baltimore, Maryland 21287.
J Neurosci. 1994 Jun;14(6):3915-21. doi: 10.1523/JNEUROSCI.14-06-03915.1994.
Slow axonal transport is the mechanism by which cytoskeletal proteins are distributed within the axon. This function has traditionally been considered an exclusively unidirectional, anterograde process. Previous observations of cytoskeletal redistribution in surviving, transected axons of the C57BL/Ola mouse led us to hypothesize a retrograde component of cytoskeletal transport. To test this hypothesis against previous methods of measuring slow transport of cytoskeleton, we radioactively pulse-labeled proteins in sensory neurons of C57BL/Ola mice and followed their redistribution by gel fluorography in ligated and unligated sciatic nerves. Slow axonal transport of cytoskeletal proteins proceeded with the same characteristics in C57BL/Ola as in standard C57BL/6 mice. In comparison to the transport profiles from unligated control nerves, in ligated nerves there was redistribution of radiolabeled neurofilament and tubulin proteins back toward the cell body during the 14 d experimental period. These observations demonstrate that pulse-labeled cytoskeletal proteins move bidirectionally in this experimental system, and may provide insight into the normal mechanisms of cytoskeletal maintenance.
慢轴突运输是细胞骨架蛋白在轴突内分布的机制。传统上,这一功能被认为是完全单向的顺行过程。先前对C57BL/Ola小鼠存活的横断轴突中细胞骨架重新分布的观察使我们推测细胞骨架运输存在逆行成分。为了用先前测量细胞骨架慢运输的方法来验证这一假设,我们用放射性脉冲标记C57BL/Ola小鼠感觉神经元中的蛋白质,并通过凝胶荧光成像追踪其在结扎和未结扎的坐骨神经中的重新分布。细胞骨架蛋白的慢轴突运输在C57BL/Ola小鼠中的特征与标准C57BL/6小鼠相同。与未结扎对照神经的运输情况相比,在结扎神经中,放射性标记的神经丝和微管蛋白在14天的实验期内会重新分布回细胞体。这些观察结果表明,脉冲标记的细胞骨架蛋白在该实验系统中双向移动,可能有助于深入了解细胞骨架维持的正常机制。