Cid-Arregui A, Parton R G, Simons K, Dotti C G
Cell Biology Program, European Molecular Biology Laboratory, Heidelberg, Germany.
J Neurosci. 1995 Jun;15(6):4259-69. doi: 10.1523/JNEUROSCI.15-06-04259.1995.
The envelope glycoproteins of Semliki Forest virus (SFV), Vesicular Stomatitis virus (VSV), and Influenza Fowl Plague virus (FPV) are vectorially targeted in neurons to the plasma membrane of dendrites (SFV and VSV) and axons (FPV). To gain insight into the mechanisms responsible for such polarized delivery we have examined the effects on neurons of nocodazole and brefeldin A (BFA), which are known to cause microtubule depolymerization and disassembly of the Golgi apparatus, respectively. Nocodazole treatment blocked transport of all viral glycoproteins to both axons and dendrites. BFA treatment induced disruption of the Golgi complex, including the trans-Golgi network (TGN), and tubulation of endosomes. However, the delivery of the SFV and FPV glycoproteins to the cell surface was not affected significantly by BFA, although processing and sorting were altered, as revealed by surface biotinylation and immunofluorescence microscopy of fixed nonpermeabilized cells. These results demonstrate the involvement of microtubules in axonal and dendritic transport of integral membrane glycoproteins, and the existence of a BFA-sensitive component in the sorting but not in the transport machinery.
塞姆利基森林病毒(SFV)、水疱性口炎病毒(VSV)和禽流行性感冒病毒(FPV)的包膜糖蛋白在神经元中以向量方式靶向树突(SFV和VSV)和轴突(FPV)的质膜。为了深入了解这种极化递送的机制,我们研究了诺考达唑和布雷菲德菌素A(BFA)对神经元的影响,已知它们分别会导致微管解聚和高尔基体解体。诺考达唑处理阻断了所有病毒糖蛋白向轴突和树突的转运。BFA处理导致高尔基体复合物(包括反式高尔基体网络(TGN))的破坏和内体的微管化。然而,尽管通过固定的非通透细胞的表面生物素化和免疫荧光显微镜观察发现加工和分选发生了改变,但BFA对SFV和FPV糖蛋白向细胞表面的递送没有显著影响。这些结果证明了微管参与了整合膜糖蛋白的轴突和树突运输,以及在分选而非运输机制中存在对BFA敏感的成分。