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


CONCLUSIONS
Overexpression of mouse DVL-1 protein inhibits wortmannin-induced hyperphosphorylation of neurofilament and tau in N2a cells.
Wang
Hai-hong
HH
Department of Pathophysiology, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China.
Zhang
Ai-hong
AH
Zhu
Ling-qiang
LQ
Wang
Qun
Q
Wang
Jian-zhi
JZ
D048868:Adaptor Proteins, Signal Transducing; D000730:Androstadienes / Q000494:pharmacology; D000818:Animals; D000072261:Dishevelled Proteins; D051379:Mice; D009447:Neuroblastoma / Q000378:metabolism* / Q000473:pathology; D016900:Neurofilament Proteins / Q000378:metabolism*; D010750:Phosphoproteins; D010766:Phosphorylation / Q000187:drug effects; D011506:Proteins / Q000235:genetics / Q000378:metabolism* / Q000502:physiology; D014162:Transfection; D014407:Tumor Cells, Cultured; D000077191:Wortmannin; D016875:tau Proteins / Q000378:metabolism*
D048868:Adaptor Proteins, Signal Transducing; D000730:Androstadienes; D000072261:Dishevelled Proteins; C000606579:Dvl1 protein, mouse; D016900:Neurofilament Proteins; D010750:Phosphoproteins; D011506:Proteins; D016875:tau Proteins; D000077191:Wortmannin