The neurotoxin, MPP+, induces hyperphosphorylation of Tau, in the presence of alpha-Synuclein, in SH-SY5Y neuroblastoma cells.
Alzheimer's disease (AD) is characterized, in part, by intracellular neurofibrillary tangles composed of hyperphosphorylated filamentous aggregates of the microtubule-associated protein, Tau. Such hyperphosphorylated Tau is also found in Lewy bodies (LBs), and cytoplasmic inclusion bodies in certain forms of Parkinson's disease (PD). Further, LBs also contain aggregates of alpha-synuclein (alpha-Syn), also a microtubule-associated protein, which has been linked to the genesis of PD. To investigate a specific correlation between Tau phosphorylation and alpha-Syn, we generated a SH-SY5Y cell line that stably expresses human wild type alpha-Syn. Protein expression levels in the stably transfected cell line (SHalpha-Syn) were within the physiological range of alpha-Syn expression found in Substantia nigra. We show here, in the MPP+ (1-methyl-4-phenylpyridinium ion) cell model of parkinsonism, a time- and dose-dependent increase in the hyperphosphorylation of Tau at pSer396/404 (PHF-1-reactive Tau, p-Tau), concomitant with increased accumulation of alpha-Syn, upon treatment of cells with the neurotoxin. This increase in p-Tau was strictly dependent on the presence of alphaSyn, since in transfected cells not expressing any alpha-Syn, MPP+ failed to induce an increase in PHF-1-reactive Tau. The production of p-Tau caused increased cytotoxicity as indexed by reduced cell viability. Moreover, in the absence of alpha-Syn, the cells were more resistant to MPP+ -induced cell death. The increased levels of both p-Tau and alpha-Syn led to diminished levels of these proteins associated with the cytoskeleton, which was accompanied by enhanced presence of the proteins in the cytoskeletal-free fractions. These data indicate that alpha-Syn and p-Tau modulate the pathogenicity of one another, suggesting a novel convergent mechanism of neurodegeneration.
Duka
Tetyana
T
Department of Biochemistry and Molecular AND Cellular Biology, Georgetown University, Washington DC 20007, USA.
Sidhu
Anita
A
D015655:1-Methyl-4-phenylpyridinium / Q000494:pharmacology*; D015153:Blotting, Western / Q000379:methods; D002458:Cell Fractionation / Q000379:methods; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D050483:Dopamine Plasma Membrane Transport Proteins / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D015870:Gene Expression / Q000187:drug effects*; D006801:Humans; D009447:Neuroblastoma; D009498:Neurotoxins / Q000494:pharmacology*; D010766:Phosphorylation / Q000187:drug effects; D012694:Serine / Q000378:metabolism; D013778:Tetrazolium Salts; D013844:Thiazoles; D014162:Transfection / Q000379:methods; D051844:alpha-Synuclein / Q000235:genetics / Q000378:metabolism*; D016875:tau Proteins / Q000378:metabolism*
D050483:Dopamine Plasma Membrane Transport Proteins; C054369:MAPT protein, human; D009498:Neurotoxins; D013778:Tetrazolium Salts; D013844:Thiazoles; D051844:alpha-Synuclein; D016875:tau Proteins; D012694:Serine; C022616:thiazolyl blue; D015655:1-Methyl-4-phenylpyridinium