Open AccessOverexpression of dishevelled‐1 attenuates wortmannin‐induced hyperphosphorylation of cytoskeletal proteins in N2a cell 1
Author(s) -
WANG Haihong,
ZHANG Aihong,
ZHU Lingqiang,
WANG Qun,
WANG Jianzhi
Publication year - 2005
Publication title -
acta pharmacologica sinica
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.514
H-Index - 90
eISSN - 1745-7254
pISSN - 1671-4083
DOI - 10.1111/j.1745-7254.2005.00131.x
Subject(s) - wortmannin , neurofilament , hyperphosphorylation , western blot , phosphorylation , cytoskeleton , microbiology and biotechnology , epitope , chemistry , transfection , biology , biochemistry , cell , immunology , antibody , protein kinase b , immunohistochemistry , gene
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 TM 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 μmol/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 atPHF‐1 (Ser‐396/404), M4 (Thr‐23 1/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.