在N2a细胞中，Dishevelled-1的过表达可减弱渥曼青霉素诱导的细胞骨架蛋白的过度磷酸化。

Overexpression of dishevelled-1 attenuates wortmannin-induced hyperphosphorylation of cytoskeletal proteins in N2a cell.

作者信息

Wang Hai-hong, Zhang Ai-hong, Zhu Ling-qiang, Wang Qun, Wang Jian-zhi

机构信息

Department of Pathophysiology, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China.

出版信息

Acta Pharmacol Sin. 2005 Jun;26(6):679-84. doi: 10.1111/j.1745-7254.2005.00131.x.

DOI:10.1111/j.1745-7254.2005.00131.x

PMID:15916733

Abstract

AIM

To investigate the effect of dishevelled-1 (DVL-1) on wortmannin-induced Alzheimer-like hyperphosphorylation of cytoskeletal proteins in mouse neuroblastoma 2a (N2a) cells.

METHODS

Cultured N2a cells were transitorily transfected with DVL-1 expression plasmid using Lipofectamine 2000. Western blot and immunofluorescence microscopy were used to measure the phosphorylation of neurofilament and tau.

RESULTS

Level of phosphorylated neurofilament at SMI31 epitope and phosphorylated tau determined by PHF-1 was increased at 1 h and 3 h and back to normal at 6 h after wortmannin 1 micromol/L treatment. The highest level of phosphorylated neurofilament and phosphorylated tau was seen at 1 h and 3 h after wortmannin treatment, respectively. When DVL-1 protein was overexpressed, the hyperphosphorylation of neurofilament at SMI31 and SMI32 epitopes and tau at PHF-1 (Ser-396/404), M4 (Thr-231/Ser-235), and Tau-1 (Ser-198/199/202) epitopes was attenuated.

CONCLUSION

Overexpression of mouse DVL-1 protein inhibits wortmannin-induced hyperphosphorylation of neurofilament and tau in N2a cells.

摘要

目的

研究蓬乱蛋白-1（DVL-1）对渥曼青霉素诱导的小鼠神经母细胞瘤2a（N2a）细胞细胞骨架蛋白阿尔茨海默样过度磷酸化的影响。

方法

使用Lipofectamine 2000将DVL-1表达质粒瞬时转染至培养的N2a细胞。采用蛋白质免疫印迹法和免疫荧光显微镜检测神经丝和tau蛋白的磷酸化水平。

结果

1 μmol/L渥曼青霉素处理后1小时和3小时，由SMI31表位决定的神经丝磷酸化水平和由PHF-1测定的tau蛋白磷酸化水平升高，6小时后恢复正常。渥曼青霉素处理后1小时和3小时分别观察到神经丝磷酸化和tau蛋白磷酸化的最高水平。当DVL-1蛋白过表达时，SMI31和SMI32表位的神经丝过度磷酸化以及PHF-1（Ser-396/404）、M4（Thr-231/Ser-235）和Tau-1（Ser-198/199/202）表位的tau蛋白过度磷酸化减弱。