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蛋白质在迁移性小脑颗粒细胞质膜中的正向转运。

Forward transport of proteins in the plasma membrane of migrating cerebellar granule cells.

机构信息

Institute of Neuroscience, State Key Laboratory of Neuroscience, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200031, China.

出版信息

Proc Natl Acad Sci U S A. 2012 Dec 18;109(51):E3558-67. doi: 10.1073/pnas.1219203110. Epub 2012 Dec 3.

Abstract

Directional flow of membrane components has been detected at the leading front of fibroblasts and the growth cone of neuronal processes, but whether there exists global directional flow of plasma membrane components over the entire migrating neuron remains largely unknown. By analyzing the trajectories of antibody-coated single quantum dots (QDs) bound to two membrane proteins, overexpressed myc-tagged synaptic vesicle-associated membrane protein VAMP2 and endogenous neurotrophin receptor TrkB, we found that these two proteins exhibited net forward transport, which is superimposed upon Brownian motion, in both leading and trailing processes of migrating cerebellar granule cells in culture. Furthermore, no net directional transport of membrane proteins was observed in nonmigrating cells with either growing or stalling leading processes. Analysis of the correlation of motion direction between two QDs on the same process in migrating neurons also showed a higher frequency of correlated forward than rearward movements. Such correlated QD movements were markedly reduced in the presence of myosin II inhibitor blebbistatin,suggesting the involvement of myosin II-dependent active transport processes. Thus, a net forward transport of plasma membrane proteins exists in the leading and trailing processes of migrating neurons, in line with the translocation of the soma.

摘要

膜成分的定向流动已在成纤维细胞的前沿和神经元突起的生长锥中被检测到,但整个迁移神经元的质膜成分是否存在整体定向流动在很大程度上仍是未知的。通过分析两种膜蛋白(过表达的 myc 标记突触小泡相关膜蛋白 VAMP2 和内源性神经营养因子受体 TrkB)结合的抗体包被的单量子点(QDs)的轨迹,我们发现这两种蛋白在前导和尾随突起中都表现出净向前运输,这种运输叠加在布朗运动之上,在培养的小脑颗粒细胞的迁移过程中。此外,在具有生长或停滞前导过程的非迁移细胞中,没有观察到膜蛋白的净定向运输。对迁移神经元中同一突起上两个 QD 运动方向相关性的分析也显示,向前运动的相关性频率高于向后运动。在肌球蛋白 II 抑制剂 blebbistatin 的存在下,这种相关的 QD 运动明显减少,表明肌球蛋白 II 依赖性主动运输过程的参与。因此,在迁移神经元的前导和尾随突起中存在质膜蛋白的净向前运输,与体的易位一致。

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