KIF1A is the primary anterograde motor protein required for the axonal transport of dense-core vesicles in cultured hippocampal neurons
KIF1A is the primary anterograde motor protein required for the axonal transport of dense-core vesicles in cultured hippocampal neurons
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DOI：
10.1016/j.neulet.2011.01.018
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发表时间：
2011-03-24
影响因子：
2.5
通讯作者：
Silverman, M. A.
中科院分区：
文献类型：
--
作者：
Lo, K. Y.;Kuzmin, A.;Silverman, M. A.
致密核囊泡（DCV）负责运输、加工和分泌神经肽货物，所述神经肽货物介导广泛的生物过程，包括神经元发育、存活以及学习和记忆。DCV在细胞体中合成，并由驱动蛋白马达蛋白沿着微管转运到突触前和突触后释放位点。由于依赖于基于驱动蛋白的转运，我们试图确定驱动蛋白-3家族成员KIF 1A是否在原代培养的海马神经元中转运DCV，如已描述的无脊椎动物神经元。双色活细胞成像显示，DCV标记物嗜铬粒蛋白A-RFP和BDNF-RFP与KIF 1A-GFP一起在顺行和逆行方向上移动。为了证明KIF 1A在DCV转运中的功能作用，使用RNA干扰（shRNA）降低神经元中的运动蛋白表达。双标记的DCV标记物显示在表达针对KIF 1A的shRNA的细胞中细胞器通量显著减少。由KIF 1A以外的马达驱动的货物运输，包括线粒体和转铁蛋白受体，在KIF 1A shRNA表达细胞中不受影响。总之，这些数据支持KIF 1A在哺乳动物神经元中DCV的顺行运输中的主要作用，并且还提供了KIF 1A在逆行DCV运输期间与DCV保持相关的证据。皇冠版权所有（C）2011由爱思唯尔爱尔兰有限公司出版。保留所有权利。
Dense-core vesicles (DCVs) are responsible for transporting, processing, and secreting neuropeptide cargos that mediate a wide range of biological processes, including neuronal development, survival, and learning and memory. DCVs are synthesized in the cell body and are transported by kinesin motor proteins along microtubules to pre- and postsynaptic release sites. Due to the dependence on kinesin-based transport, we sought to determine if the kinesin-3 family member, KIF1A, transports DCVs in primary cultured hippocampal neurons, as has been described for invertebrate neurons. Two-color, live-cell imaging showed that the DCV markers, chromogranin A-RFP and BDNF-RFP, move together with KIF1A-GFP in both the anterograde and retrograde directions. To demonstrate a functional role for KIF1A in DCV transport, motor protein expression in neurons was reduced using RNA interference (shRNA). Fluorescently tagged DCV markers showed a significant reduction in organelle flux in cells expressing shRNA against KIF1A. The transport of cargo driven by motors other than KIF1A, including mitochondria and the transferrin receptor, was unaffected in KIF1A shRNA expressing cells. Taken together, these data support a primary role for KIF1A in the anterograde transport of DCVs in mammalian neurons, and also provide evidence that KIF1A remains associated with DCVs during retrograde DCV transport. Crown Copyright (C) 2011 Published by Elsevier Ireland Ltd. All rights reserved.