KINESIN ASSOCIATES WITH ANTEROGRADELY TRANSPORTED MEMBRANOUS ORGANELLES INVIVO
KINESIN ASSOCIATES WITH ANTEROGRADELY TRANSPORTED MEMBRANOUS ORGANELLES INVIVO
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DOI：
10.1083/jcb.114.2.295
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发表时间：
1991-07-01
影响因子：
7.8
通讯作者：
BRADY, ST
中科院分区：
文献类型：
--
作者：
HIROKAWA, N;SATOYOSHITAKE, R;BRADY, ST
生化、药理学和免疫细胞化学研究表明，微管激活的 ATP 酶、驱动蛋白参与快速轴突运输中膜结合细胞器的运动。体外研究表明，驱动蛋白优先沿顺行方向移动细胞器，但缺乏有关驱动蛋白在活轴突中的功能和精确定位的数据。目前的研究旨在确定驱动蛋白是否与体内顺行或逆行移动细胞器相关。周围神经被结扎以产生沿确定方向移动的细胞器的积累。通过免疫荧光和使用超低温切片术的免疫金电子显微镜分析连接的近端（顺行）和远端（逆行）区域的驱动蛋白定位。大量的驱动蛋白与近侧的顺行移动细胞器相关，而在远端检测到的驱动蛋白明显较少。统计分析表明驱动蛋白主要与膜结合的细胞器相关。这些观察结果表明轴突驱动蛋白主要与体内顺行移动细胞器相关。
Biochemical, pharmacological and immunocytochemical studies have implicated the microtubule-activated ATPase, kinesin, in the movement of membrane bounded organelles in fast axonal transport. In vitro studies suggested that kinesin moves organelles preferentially in the anterograde direction, but data about the function and precise localization of kinesin in the living axon were lacking. The current study was undertaken to establish whether kinesin associates with anterograde or retrograde moving organelles in vivo. Peripheral nerves were ligated to produce accumulations of organelles moving in defined directions. Regions proximal (anterograde) and distal (retrograde) to the ligation were analyzed for kinesin localization by immunofluorescence, and by immunogold electron microscopy using ultracryomicrotomy. Substantial amounts of kinesin were associated with anterograde moving organelles on the proximal side, while significantly less kinesin was detected distally. Statistical analyses indicated that kinesin was mostly associated with membrane-bounded organelles. These observations indicate that axonal kinesin is primarily associated with anterograde moving organelles in vivo.