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 LipofectamineTM 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.
Similar content being viewed by others
Article PDF
References
Iqbal K, Grundke-Iqbal I, Smith AJ, George L, Tung YC, Zaidi T . Identification and localization of a tau peptide to paired helical filaments of Alzheimer disease. Proc Natl Acad Sci USA 1989; 86: 5646–50.
Wang JZ, Tung YC, Wang YP, Li XT, Iqbal K, Grundke-Iqbal I . Hyperphosphorylation and accumulation neuro filament proteins in Alzheimer disease brain and in okadaic acid-treated SY5Y cells. FEBS Lett 2001; 507: 81–7.
Gong CX, Wang JZ, Iqbal K, Grundke-Iqbal I . Inhibition of protein phosphatase 2A induces phosphorylation and accumulation of neuro filaments in metabolically active rat brain slices. Neurosci Lett 2003; 340: 107–10.
Wang JZ, Wu Q, Smith A, Grundke-Iqbal I, Iqbal K . Tau is phosphorylated by GSK-3 at several sites found in Alzheimer disease and its biological activity markedly inhibited only after it is prephosphorylated by A-kinase. FEBS Lett 1998; 436: 28–34.
Liu SJ, Zhang JY, Li HL, Fang ZY, Wang Q, Deng HM, et al. Tau becomes a more favorable substrate for GSK-3 when it is prephosphorylated by PKA in rat brain. J Biol Chem 2004; 279: 50078–88.
Pei JJ, Tanaka T, Tung YC, Braak E, Iqbal K, Grundke-Iqbal I . Distribution, levels, and activity of glycogen synthase kinase-3 in the Alzheimer disease brain. J Neuropathol Exp Neurol 1997; 56: 70–8.
Guan RJ, Khatra BS, Cohlberg JA . Phosphorylation of bovine neurofilament proteins by protein kinase FA (glycogen synthase kinase 3). J Biol Chem 1991; 266: 8262–7.
Guidato S, Tsai LH, Woodgett J, Miller CC . Differential cellular phosphorylation of neurofilament heavy side-arms by glycogen synthase kinase-3 and cyclin-dependent kinase-5. J Neurochem 1996; 66: 1698–706.
Cross DA, Alessi DR, Cohen P, Andjelkovich M, Hemmings BA . Inhibition of glycogen synthase kinase-3 by insulin mediated by protein kinase B. Nature 1995; 378: 785–9.
Welsh GI, Foulstone EJ, Young SW, Tavare JM, Proud CG . Wortmannin inhibits the effects of insulin and serum on the activities of glycogen synthase linase-3 and mitogen-activated protein kinase. Biochem J 1994; 303: 15–20.
Liu SJ, Wang JZ . Alzheimer-like tau phosphorylation induced by wortmannin in vivo and its attenuation by melatonin. Acta Pharmacol Sin 2002; 23: 183–7.
Liu SJ, Zhang AH, Li HL, Wang Q, Deng HM, Netzer WJ, et al. Overactivation of glycogen synthase kinase-3 by inhibition of phosphoinositol-3 kinase and protein kinase C leads to hyperphosphorylation of tau and impairment of spatial memory. J Neurochem 2003; 87: 1333–44.
Klingensmith, J Nusse R, Perrimon N . The Drosophila segment polarity gene dishevelled encodes a novel protein required for response to the wingless signal. Genes Dev 1994; 8: 118–30.
Wagner U, Brownlees J, Irving NG, Lucas FR, Salinas PC, Miller CC . Overexpression of the mouse dishevelled-1 protein inhibits GSK-3β-mediated phosphorylation of tau in transfected mammalian cells. FEBS Lett 1997; 411: 369–72.
Mudher A, Chapman S, Richardson J, Asuni A, Gibb G, Pollard C, et al. Dishevelled regulates the metabolism of amyloid precursor protein via protein kinase C/mitogen-activated protein kinase and c-Jun terminal kinase. J Neurosci 2001; 21: 4987–95.
Yu J, Deng YQ, Yang Y, Zhang JY, Zhang YP, Zhang SH, et al. Activation of glycogen synthase kinase 3 induces Alzheimer-like hyperphosphorylation of cytoskeleton protein and cell damage. Prog Biochem Biophys 2004; 31: 532–7.
Andreadis A, Broderick JA, Kosik KS . Relative exon affinities and suboptimal splice site signals lead to non-equivalence of two cassette exons. Nucleic Acids Res 1995; 23: 3585–93.
Cook D, Fry MJ, Hughes K, Sumathipala R, Woodgett JR, Dale TC . Wingless inactivates glycogen synthase kinase-3 via an intracellular signalling pathway which involves a protein kinase C. EMBO J 1996; 15: 4526–36.
Alvarez AR, Godoy JA, Mullendorff K, Olivares GH, Bronfman M, Inestrosa NC . Wnt-3a overcomes beta-amyloid toxicity in rat hippocampal neurons. Exp Cell Res 2004; 297: 186–96.
Yanagawa S, Van Leeuwen F, Wodarz A, Klingensmith J, Nusse R . The dishevelled protein is modified by wingless signaling in Drosophila. Genes Dev 1995; 9: 1087–97.
Author information
Authors and Affiliations
Corresponding author
Additional information
Project supported in part by grants from the National Natural Science Foundation of China (No 30170221 and No 30430270).
Rights and permissions
About this article
Cite this article
Wang, Hh., Zhang, Ah., Zhu, Lq. et al. Overexpression of dishevelled-1 attenuates wortmannin-induced hyperphosphorylation of cytoskeletal proteins in N2a cell. Acta Pharmacol Sin 26, 679–684 (2005). https://doi.org/10.1111/j.1745-7254.2005.00131.x
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1111/j.1745-7254.2005.00131.x