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Cytoskeletons and cell death-associated proteins expressed in neuronal cells in the neurofilament gene-manipulated mice

神经丝基因操作小鼠神经细胞中表达的细胞骨架和细胞死亡相关蛋白

基本信息

批准号：
14570030
负责人：
GOTOW Takahiro
金额：
$ 2.24万
依托单位：
Koshien University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2002
资助国家：
日本
起止时间：
2002 至 2003
项目状态：
已结题

项目摘要

Neurofilaments (NFs) are composed of three proteins, NF-L,NF-M and NP-H, and NF-M and NP-H are provided with uniquely extended and phosphorylated carboxyl-terminal tail domains that do not appear in other intermediate filament proteins. In collaboration with American researchers at California (UCSD) and New York Universities we generated knockout mice that do not express each of these subunit proteins as well as knockin mice that lack only the carboxyl-terminal tail of NF-M or/and NF-H, to know how the deletions of each NF subunit protein and of the extended carboxy-terminal tails of NF-M or/and NF-H are related with NF organization in the neuron, axonal structure, axonal transport of NFs, organizations of other cytoskeletons and membrane-bound organelles, and expression of other NF subunit proteins. When NF-L was deleted, NFs were not assembled in neurons. Such axons were much thinner but provided with many more microtubules and mitochondria. Compared with NF-H, the deletion of NF-M had more effective influence on axonal calibers and NF organization, suggesting that NF-M is more essential for the NF function. Since the carboxyl-terminal tail of NF-M appears to be more important for NF organization and function, we deleted only this tail domain of NF-M or/and NF-H, and found as expected that NF-M tail was more necessary for the NF organization. However, more drastic changes in axoplasmic organizations, such as irregularity of NF alignment, Increase in density of microtubules as well as mitochondria, were obtained when both NF-M and NF-H tails were simultaneously deleted. These results suggest that both NF-M and NP-H tails, which form crossbridges between NFs, are necessary for NF organization and axonal functions, and are also associated with regulation of neuronal cell death through the change in function of mitochondria.

神经丝（nf）由NF-L、NF-M和NP-H三种蛋白组成，NF-M和NP-H具有独特的延伸和磷酸化的羧基末端尾部结构域，这在其他中间丝蛋白中是不存在的。在与加州大学圣地亚哥分校和纽约大学的美国研究人员的合作中，我们产生了不表达这些亚基蛋白的敲除小鼠，以及只缺乏NF- m或/和NF- h的羧基末端尾部的敲除小鼠，以了解每个NF亚基蛋白的缺失以及NF- m或/和NF- h的延伸羧基末端尾部的缺失如何与NF在神经元中的组织，轴突结构，NF的轴突运输，其他细胞骨架和膜结合细胞器的组织，以及其他NF亚基蛋白的表达。NF-L被删除后，神经元中nf不再组装。这样的轴突要薄得多，但具有更多的微管和线粒体。与NF- h相比，NF- m的缺失对NF轴突直径和NF组织的影响更大，表明NF- m对NF功能更重要。由于NF- m的羧基末端尾部似乎对NF的组织和功能更重要，因此我们只删除NF- m或/和NF- h的这一尾部结构域，并发现NF- m尾部对NF的组织更为必要。然而，当NF- m和NF- h尾巴同时被删除时，轴浆组织发生了更剧烈的变化，如NF排列不规则，微管密度增加以及线粒体。这些结果表明，NF- m和NP-H尾在NF之间形成交叉桥，是NF组织和轴突功能所必需的，并且通过线粒体功能的改变与神经元细胞死亡的调节有关。