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通过显微注射在体外研究背根神经节单个神经元中辣根过氧化物酶的轴浆运输。

Axoplasmic transport of horseradish peroxidase in single neurons of the dorsal root ganglion studied in vitro by microinjection.

作者信息

Meller K

机构信息

Abteilung für Cytologie, Ruhr-Universität Bochum, Federal Republic of Germany.

出版信息

Cell Tissue Res. 1992 Oct;270(1):139-48. doi: 10.1007/BF00381888.

Abstract

The dependence of anterograde axoplasmic transport on cytoskeletal components was investigated using microinjection of horseradish peroxidase (HRP) into the somata of chick dorsal root ganglion cells in vitro. Microinjected HRP was transported anterogradely in the neurites and their branches; this transport was disturbed by colchicine in a drug-dependent and time-dependent manner. Cytochalasin B, a drug that depolymerizes actin, did not inhibit the transport of HRP, despite the formation of local swellings in neurites. The microinjection of polyclonal antibodies directed against tubulin and monoclonal antibodies (mAbs) against 200-kDa neurofilaments disturbed the axoplasmic transport of co-injected HRP, which then exhibited an irregular and discontinuous distribution in the axonal branches. The transport of HRP became discontinuous after the injection of anti-tubulin antibodies and led to the formation of globular deposits of HRP. Polyclonal antibodies against actin and mAbs to 160-kDa and 68-kDa neurofilaments seemed to have no effect on the axoplasmic transport of co-injected HRP. Microinjection of antibodies against tubulin induced formation of perinuclear bundles consisting of cytoskeletal components. The transport of HRP thus appears to be regulated by an intact microtubular system and cross-linker components (200-kDa neurofilaments) of the cytoskeleton. Actin and most intermediate filament proteins do not seem to play an essential role in the transport of HRP.

摘要

利用体外将辣根过氧化物酶(HRP)显微注射到鸡背根神经节细胞的胞体中,研究了顺行轴浆运输对细胞骨架成分的依赖性。显微注射的HRP在神经突及其分支中顺行运输;这种运输受到秋水仙碱的干扰,且呈药物依赖性和时间依赖性。细胞松弛素B是一种能使肌动蛋白解聚的药物,尽管神经突中形成了局部肿胀,但它并未抑制HRP的运输。针对微管蛋白的多克隆抗体和针对200-kDa神经丝的单克隆抗体(mAb)的显微注射干扰了共注射的HRP的轴浆运输,随后HRP在轴突分支中呈现出不规则和不连续的分布。注射抗微管蛋白抗体后,HRP的运输变得不连续,并导致HRP形成球状沉积物。针对肌动蛋白的多克隆抗体以及针对160-kDa和68-kDa神经丝的mAb似乎对共注射的HRP的轴浆运输没有影响。针对微管蛋白的抗体显微注射诱导了由细胞骨架成分组成的核周束的形成。因此,HRP的运输似乎受细胞骨架完整的微管系统和交联成分(200-kDa神经丝)的调节。肌动蛋白和大多数中间丝蛋白似乎在HRP的运输中不发挥重要作用。

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