Assembly and Maintenance of the Axonal Cytoskeleton in Cultured Neurons

近年来，一些新的神经元的存在， 中间丝物种已被记录。 谢伊医生 显示“66 kDa神经元中间丝”，也称为 作为α-介连蛋白，由NB 2a/d1细胞组成型表达， 并在神经炎基底内显示出独特的分布， 轴突丘在分化过程中。 谢伊医生会调整这个 在不同分化阶段的细胞中， 已经用于轴突细胞骨架研究的方法 动力学：1）抑制易位，组装，和 这种神经元中间丝种类与 其他细胞骨架成分，和2）抑制合成 通过用反义寡核苷酸处理细胞来研究这种蛋白质。 每隔一小时将通过相衬检查细胞 显微镜检查神经突生长的变化，神经突 收缩，稳定性受损，以及免疫细胞化学， 66 kDa神经元中间体的细胞内定位 丝/α-互联蛋白以及其他轴突细胞骨架 抗原 这些分析将有助于确定NF-66是否 是神经突生长、持续延长或 稳定化 以下结果的比较 下调66 kDa神经元中间体 丝/α-interexin与H-200将提供信息 关于这种新的神经元中间体所起的作用， 轴突生长和成熟中的丝状体种类。

In recent years, the presence of several novel neuronal intermediate filament species has been documented. Dr. Shea has shown that the "66kDa neuronal intermediate filament," also known as alpha-internexin, is constitutively expressed by NB2a/d1 cells, and displays a unique distribution within the neuritic base and axon hillock during differentiation. Dr. Shea will modulate this protein in cells at various stages of differentiation by two methods already in use for the study of axonal cytoskeletal dynamics: 1) inhibiting the translocation, assembly, and interaction of this neuronal intermediate filament species with other cytoskeletal components, and 2) inhibiting the synthesis of this protein by treating cells with antisense oligonucleotides. Cells will be examined at hourly intervals by phase-contrast microscopy for alterations in neurite outgrowth, neurite retraction, compromise of stability, and immunocytochemistry for intracellular localization of the 66kDa neuronal intermediate filament/alpha-internexin as well as other axonal cytoskeletal antigens. These analyses will help determine whether or not NF-66 is required for neurite outgrowth, continued elongation, or stabilization. Comparison of results obtained following down-regulation of the 66kDa neuronal intermediate filament/alpha-internexin with H-200 will provide information regarding the role played by this novel neuronal intermediate filament species in axonal outgrowth and maturation.