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预处理性神经挤压可加速后续挤压后形成的新芽中细胞骨架蛋白的运输。

Conditioning nerve crush accelerates cytoskeletal protein transport in sprouts that form after a subsequent crush.

作者信息

McQuarrie I G, Jacob J M

机构信息

Department of Neurosurgery, School of Medicine, Case Western Reserve University, Cleveland, Ohio 44106.

出版信息

J Comp Neurol. 1991 Mar 1;305(1):139-47. doi: 10.1002/cne.903050113.

Abstract

To examine the relationship between axonal outgrowth and the delivery of cytoskeletal proteins to the growing axon tip, outgrowth was accelerated by using a conditioning nerve crush. Because slow component b (SCb) of axonal transport is the most rapid vehicle for carrying cytoskeletal proteins to the axon tip, the rate of SCb was measured in conditioned vs. sham-conditioned sprouts. In young Sprague-Dawley rats, the conditioning crush was made to sciatic nerve branches at the knee; 14 days later, the test crush was made where the L4 and L5 spinal nerves join to form the sciatic nerve in the flank. Newly synthesized proteins were labeled in motor neurons by injecting 35S-methionine into the lumbar spinal cord 7 days before the test crush. The wave of pulse-labeled SCb proteins reached the crush by the time it was made and subsequently entered sprouts. The nerve was removed and sectioned for SDS-PAGE and fluorography 4-12 days after the crush. Tubulins, neurofilament proteins, and representative "cytomatrix" proteins (actin, calmodulin, and putative microtubule-associated proteins) were removed from gels for liquid scintillation counting. Labeled SCb proteins entered sprouts without first accumulating in parent axon stumps, presumably because sprouts begin to grow within hours after axotomy. The peak of SCb moved 11% faster in conditioned than in sham-conditioned sprouts: 3.0 vs. 2.7 mm/d (p less than 0.05). To confirm that sprouts elongate more rapidly when a test crush is preceded by a conditioning crush, outgrowth distances were measured in a separate group of rats by labeling fast axonal transport with 3H-proline 24 hours before nerve retrieval.(ABSTRACT TRUNCATED AT 250 WORDS)

摘要

为研究轴突生长与细胞骨架蛋白向生长中的轴突末端运输之间的关系,通过进行预处理性神经挤压来加速轴突生长。由于轴突运输的慢成分b(SCb)是将细胞骨架蛋白运输到轴突末端的最快载体,因此在预处理组与假预处理组的新芽中测量了SCb的运输速率。在幼年Sprague-Dawley大鼠中,在膝部对坐骨神经分支进行预处理性挤压;14天后,在侧腹处对L4和L5脊神经汇合形成坐骨神经的部位进行测试性挤压。在测试性挤压前7天,通过向腰脊髓注射35S-甲硫氨酸,在运动神经元中标记新合成的蛋白质。脉冲标记的SCb蛋白在进行测试性挤压时已到达挤压部位并随后进入新芽。挤压后4至12天,取出神经进行切片,用于SDS-聚丙烯酰胺凝胶电泳(SDS-PAGE)和荧光自显影。从凝胶中分离出微管蛋白、神经丝蛋白和代表性的“细胞基质”蛋白(肌动蛋白、钙调蛋白和假定的微管相关蛋白)用于液体闪烁计数。标记的SCb蛋白进入新芽时并未首先在母轴突残端积累,推测是因为轴突切断后数小时内新芽就开始生长。预处理组新芽中SCb的峰值移动速度比假预处理组快11%:分别为3.0 vs. 2.7 mm/天(p<0.05)。为证实预处理性挤压后测试性挤压时新芽伸长更快,在另一组大鼠中,通过在取神经前24小时用3H-脯氨酸标记快速轴突运输来测量生长距离。(摘要截短于250字)

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