Chadan S, Le Gall J Y, Di Giamberardino L, Filliatreau G
INSERM U.334, Service Hospitalier Frédéric Joliot, Département de Biologie, C.E.A., Orsay.
J Neurosci Res. 1994 Oct 1;39(2):127-39. doi: 10.1002/jnr.490390203.
Slow axonal transport of peripherin has been studied in the motor axons of both intact and regenerating rat sciatic nerves 7 days post-crush. The studies were done by two-dimensional gel electrophoresis after intraspinal injection of 35S-methionine. In the first experiment, the sciatic nerves were removed 3 weeks after the radiolabeling pulse and cut into 6 mm segments. Each nerve segment was submitted to two-dimensional gel electrophoresis and analyzed by an original procedure which allowed us to study the distribution along the nerve of the radioactivity associated with several proteins of the cytoskeleton, especially the intermediate filament proteins, peripherin, and the low molecular mass neurofilament protein, NF-L. Peripherin was transported at two main rates: 66% of the total radiolabeled peripherin moved at 1.42 mm/day and the remainder moved at 2.28 mm/day. The radioactivity associated with NF-L exhibited a similar pattern. In the second experiment, similar intraspinal injections were made 7 days after a unilateral crush of the sciatic nerve. Regenerating nerves exhibited a clear SCa wave. However, in contrast to the intact nerves, the SCb wave could not be precisely defined in the regenerating nerves. Thus, the changes in the amount of transported proteins were analyzed in the SCa wave only. Autoradiograms of 2D-PAGE revealed that in the regenerating axons, the quantity of transported peripherin in SCa was increased by 3.5-fold. In contrast, the quantity of transported NF-L was decreased by 1.6-fold. The regenerating motor axons conveyed significantly greater (approximately twofold) amounts of labeled tubulins and actin than did intact motor axons. Our results suggest that peripherin, although mainly conveyed by SCa, plays a role during the elongation process in addition to actin and tubulin.
在大鼠坐骨神经挤压伤后7天，研究了外周蛋白在完整和再生运动轴突中的慢速轴突运输。通过脊髓内注射35S-甲硫氨酸后进行二维凝胶电泳来开展这些研究。在第一个实验中，放射性标记脉冲3周后取出坐骨神经，切成6毫米的片段。每个神经片段都进行二维凝胶电泳，并通过一种原始程序进行分析，该程序使我们能够研究与几种细胞骨架蛋白相关的放射性在神经中的分布，特别是中间丝蛋白、外周蛋白和低分子量神经丝蛋白NF-L。外周蛋白以两种主要速率运输：总放射性标记外周蛋白的66%以1.42毫米/天的速度移动，其余的以2.28毫米/天的速度移动。与NF-L相关的放射性呈现出类似的模式。在第二个实验中，在单侧坐骨神经挤压伤7天后进行类似的脊髓内注射。再生神经表现出明显的SCa波。然而，与完整神经相比，再生神经中的SCb波无法精确界定。因此，仅在SCa波中分析运输蛋白量的变化。二维聚丙烯酰胺凝胶电泳的放射自显影片显示，在再生轴突中，SCa中运输的外周蛋白量增加了3.5倍。相比之下，运输的NF-L量减少了1.6倍。再生运动轴突比完整运动轴突运输的标记微管蛋白和肌动蛋白量明显更多（约两倍）。我们的结果表明，外周蛋白虽然主要由SCa运输，但除肌动蛋白和微管蛋白外，在伸长过程中也发挥作用。
