Nogo-A inhibits necdin-accelerated neurite outgrowth by retaining necdin in the cytoplasm
Nogo-A inhibits necdin-accelerated neurite outgrowth by retaining necdin in the cytoplasm
复制标题
DOI：
10.1016/j.mcn.2009.01.009
复制
发表时间：
2009-05-01
影响因子：
3.5
通讯作者：
He, Cheng
中科院分区：
文献类型：
--
作者：
Liu, Xiujie;Wang, Yonggang;He, Cheng
Nogo-A在中枢神经系统中被认为是轴突再生的抑制剂。以往的研究主要集中在Nogo-A油在少突胶质细胞中的作用，神经元细胞内Nogo-A的作用尚不明确。为深入了解Nogo-A的生理功能，以Nogo-66为诱饵进行酵母双杂交筛选。我们发现了Nogo-66和necdin之间的一种新的相互作用。诱变分析表明，在与Nogo-66相互作用中，necdin的中心区域是必不可少的。神经组织和培养皮层神经元的共免疫沉淀进一步证实了这种相互作用。形态学证据表明Nogo-A和necdin在大鼠皮层和背根神经节神经元中高度共定位。在HEK293细胞中，Nogo-A的异位表达导致necdin从细胞核保留到细胞质。此外，在PC12细胞和培养的皮质神经元中，Nogo-A的过表达抑制了necin加速的神经突生长。同时，在稳定过表达Nogo-A的PC12细胞胞质中发现了大量的necdin。综上所述，这些数据表明Nogo-A是一种新型的necdin结合蛋白，通过将necdin隔离在细胞质中来抑制necdin加速的神经突生长。(C) 2009爱思唯尔公司版权所有。
Nogo-A has been identified ill the central nervous system as in inhibitor for axonal regeneration. Previous works have mainly focused oil Nogo-A in oligodendrocytes and the roles of neuronal intracellular Nogo-A remain elusive. To gain deep insight into the physiological functions of Nogo-A, a yeast two-hybrid screening was performed with Nogo-66 as bait. We identified a new interaction between Nogo-66 and necdin. Mutagenesis analysis revealed that the central region of necdin was indispensable for the interaction of necdin with Nogo-66. The interaction was further confirmed by co-immunoprecipitation in neural tissues and Cultured cortical neurons. Morphological evidence showed that Nogo-A and necdin highly colocalized in rat Cortical and dorsal root ganglia neurons. Ectopic expression of Nogo-A in HEK293 cells led to retention of necdin from the nucleus to the Cytoplasm. Furthermore, overexpression of Nogo-A in PC12 cells and Cultured cortical neurons inhibited necdin-accelerated neurite outgrowth. Meanwhile, necdin Was found to be significantly sequestered in the cytoplasm of PC12 cells stably overexpressing Nogo-A. Together, these data suggest that Nogo-A is a novel necdin binding protein and inhibits necdin-accelerated neuronal neurite outgrowth by sequestering necdin in the cytoplasm. (C) 2009 Elsevier Inc. All rights reserved.