Wang L J, Colella R, Roisen F J
Department of Anatomical Sciences and Neurobiology, University of Louisville School of Medicine, KY 40292, USA.]
Brain Res Dev Brain Res. 1998 Feb 10;105(2):227-39. doi: 10.1016/s0165-3806(97)00180-6.
In previous studies, we demonstrated that the exogenous ganglioside GM1 increased the complexity of the microtubular network and level of tubulin, selectively changed the distribution of microtubule associated protein-2 (MAP2) immunoreactivity from the perikarya to distal neuritic processes and increased immunogold label of MAP2 in the subplasmalemmal cytoplasm, neuritic filopodia and growth cones of Neuro-2a neuroblastoma cells. Since these areas are rich in actin filaments, our data suggested that MAP2 may be associated with microfilaments in the early stages of ganglioside-induced neuritogenesis. To determine if GM1 alters neuronal morphology by facilitating the interaction of actin and MAP2, we examined the immunolocalization of these two proteins with confocal and electron microscopy. We found that along with the redistribution of MAP2 from perikaryal to neuritic regions, there was parallel redistribution of actin. The uniform subplasmalemmal actin meshwork was disrupted in areas of processes and filopodia with a redistribution of actin to these areas in close association with MAP2. Our present results suggest that gangliosides enhance neuritogenesis by redistributing actin as well as MAP2 to processes and filopodia thereby facilitating their interaction. The association of MAP2 with actin filaments is likely to be an early step in ganglioside-mediated filopodia formation.
在先前的研究中,我们证明外源性神经节苷脂GM1增加了微管网络的复杂性和微管蛋白水平,选择性地改变了微管相关蛋白2(MAP2)免疫反应性的分布,使其从胞体转移至远端神经突,并增加了神经2a神经母细胞瘤细胞的质膜下细胞质、神经突丝状伪足和生长锥中MAP2的免疫金标记。由于这些区域富含肌动蛋白丝,我们的数据表明MAP2可能在神经节苷脂诱导的神经突发生早期与微丝相关。为了确定GM1是否通过促进肌动蛋白和MAP2的相互作用来改变神经元形态,我们用共聚焦显微镜和电子显微镜检查了这两种蛋白质的免疫定位。我们发现,随着MAP2从胞体区域重新分布到神经突区域,肌动蛋白也发生了平行的重新分布。质膜下均匀的肌动蛋白网络在神经突和丝状伪足区域被破坏,肌动蛋白重新分布到这些区域,并与MAP2紧密结合。我们目前的结果表明,神经节苷脂通过将肌动蛋白和MAP2重新分布到神经突和丝状伪足来增强神经突发生,从而促进它们的相互作用。MAP2与肌动蛋白丝的结合可能是神经节苷脂介导的丝状伪足形成的早期步骤。