Increased expression of cdk5/p25 in N2a cells leads to hyperphosphorylation and impaired axonal transport of neurofilament proteins.
Increased expression of cdk5/p25 in N2a cells leads to hyperphosphorylation and impaired axonal transport of neurofilament proteins.
复制标题
N2a 细胞中 cdk5/p25 表达增加导致神经丝蛋白过度磷酸化和轴突运输受损。
DOI：
10.1016/j.lfs.2010.02.009
复制
发表时间：
2010-03
期刊：
影响因子：
6.1
通讯作者：
陈娟
中科院分区：
文献类型：
--
作者：
陈娟
阿尔茨海默病（AD）是痴呆症的主要原因。在AD患者中，cdk5表达增加，tau和NFs磷酸化增强。本研究旨在探讨cdk5活性增加对神经丝轴突运输的影响。本研究采用分子工程方法在神经母细胞瘤N2a细胞中过表达cdk5/p25，并利用活细胞成像技术研究其对轴突转运的影响。在稳定转染的细胞中，与未转染的细胞相比，cdk5活性增加了2.5倍，这反过来导致NFs和tau在几个磷酸化位点的磷酸化急剧增加。利用延时成像技术，在过表达cdk5/p25的细胞中，瞬时转染NF-M n端连接的荧光蛋白（EGFP-NFM），捕获NFs的运输。cdk5/p25细胞的NF轴突转运速率明显减慢，在细胞体中观察到固定NF簇的积累。罗斯科维汀，一种cdk5的抑制剂，显著地逆转了轴突运输中的这种缺陷。这些结果表明，在AD受试者中发现的cdk5活性增加可能与AD的发病机制至关重要，其潜在机制是促进过度磷酸化的细胞骨架NF蛋白的积累，导致NF轴突运输的持久损害。
AIMSAlzheimer's disease (AD) is the leading cause of dementia. The increased cdk5 expression and enhanced phosphorylation of tau and NFs have been seen in AD patients. Our study aimed at investigating the effects of increased cdk5 activity on axonal transport of neurofilaments (NFs).MAIN METHODSIn this study, we used a molecular engineering approach to overexpress cdk5/p25 in neuroblastoma N2a cells and investigated the effects on axonal transport with live cell imaging techniques.KEY FINDINGSIn stably transfected cells, there was a 2.5-fold increase in cdk5 activity compared to non-transfected cells, which in turn led to a dramatic increase in phosphorylation of NFs and tau at several phosphorylation sites. Using time-lapse imaging technology, the transport of NFs was captured in the cells overexpressing cdk5/p25, which were also transiently transfected with fluorescence protein linked to the N-terminus of NF-M (EGFP-NFM). The cdk5/p25 cells displayed significantly slower rates of axonal transport of NFs, with accumulation of immobile NF clusters observed in the cell body. Roscovitine, an inhibitor of cdk5, significantly reversed this defect in axonal transport.SIGNIFICANCEThese results suggest that increased cdk5 activity found in AD subjects may be crucially related to the pathogenesis of AD via an underlying mechanism by which it promotes accumulation of excessively phosphorylated cytoskeletal NF proteins, leading to the enduring impairment of axonal transport of NFs.
登录
查看更多内容
影响因子：
2.1
作者：
N. Sternberger;L. Sternberger;J. Ulrich
通讯作者：
N. Sternberger;L. Sternberger;J. Ulrich
DOI：
10.1083/jcb.200303138
发表时间：
2003-05-12
期刊：
The Journal of cell biology
影响因子：
--
作者：
Ackerley S;Thornhill P;Grierson AJ;Brownlees J;Anderton BH;Leigh PN;Shaw CE;Miller CC
通讯作者：
Miller CC
DOI：
10.1083/jcb.126.4.1031
发表时间：
1994-08
期刊：
The Journal of cell biology
影响因子：
--
作者：
Nixon RA;Paskevich PA;Sihag RK;Thayer CY
通讯作者：
Thayer CY
影响因子：
2.9
作者：
D. Watson;K. Fittro;P. Hoffman;J. Griffin
通讯作者：
D. Watson;K. Fittro;P. Hoffman;J. Griffin
影响因子：
12.7
作者：
K. Leroy;A. Boutajangout;M. Authelet;J. Woodgett;B. Anderton;J. Brion
通讯作者：
K. Leroy;A. Boutajangout;M. Authelet;J. Woodgett;B. Anderton;J. Brion