Pro-apoptotic protein glyceraldehyde-3-phosphate dehydrogenase promotes the formation of Lewy body-like inclusions.
Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) has long been recognized as a classical glycolytic protein; however, previous studies by our group and others have demonstrated that GAPDH is a general mediator initiating one or more apoptotic cascades. Our most recent findings have elucidated that an expression of a pro-apoptotic protein GAPDH is critically regulated at the promoter region of the gene. Apoptotic signals for its subsequent aggregate formation and nuclear translocation are controlled by the respective functional domains harboured within its cDNA component. In this study, coexpression of GAPDH with either wild-type or mutant (A53T) alpha-synuclein and less likely with beta-synuclein in transfected COS-7 cells was found to induce Lewy body-like cytoplasmic inclusions. Unlike its full-length construct, the deleted mutant GAPDH construct (C66) abolished these apoptotic signals, disfavouring the formation of inclusions. The generated inclusions were ubiquitin- and thioflavin S-positive appearing fibrils. Furthermore, GAPDH coimmunoprecipitated with wild-type alpha-synuclein in this paradigm. Importantly, immunohistochemical examinations of post mortem materials from patients with sporadic Parkinson's disease revealed the colocalized profiles immunoreactive against these two proteins in the peripheral zone of Lewy bodies from the affected brain regions (i.e. locus coeruleus). Moreover, a quantitative assessment showed that about 20% of Lewy bodies displayed both antigenicities. These results suggest that pro-apoptotic protein GAPDH may be involved in the Lewy body formation in vivo, probably associated with the apoptotic death pathway.
Tsuchiya
Katsumi
K
Group on Cellular Neurobiology, Josai University, Sakado, Saitama 350-0248, Japan.
Tajima
Hisao
H
Kuwae
Toyoyasu
T
Takeshima
Takao
T
Nakano
Toshiya
T
Tanaka
Masaharu
M
Sunaga
Katsuyoshi
K
Fukuhara
Yoko
Y
Nakashima
Kenji
K
Ohama
Eisaku
E
Mochizuki
Hideki
H
Mizuno
Yoshikuni
Y
Katsube
Nobuo
N
Ishitani
Ryoichi
R
D000085:Acetates / Q000378:metabolism; D000368:Aged; D000369:Aged, 80 and over; D000818:Animals; D015153:Blotting, Western / Q000379:methods; D001921:Brain / Q000378:metabolism; D019556:COS Cells; D002449:Cell Aggregation / Q000502:physiology; D002452:Cell Count; D002522:Chlorocebus aethiops; D005260:Female; D005786:Gene Expression Regulation / Q000502:physiology*; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000378:metabolism*; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006801:Humans; D007150:Immunohistochemistry / Q000379:methods; D047468:Immunoprecipitation / Q000379:methods; D002479:Inclusion Bodies / Q000378:metabolism* / Q000648:ultrastructure; D007211:Indoles; D016631:Lewy Bodies / Q000378:metabolism* / Q000648:ultrastructure; D008297:Male; D046529:Microscopy, Electron, Transmission / Q000379:methods; D008875:Middle Aged; D016297:Mutagenesis, Site-Directed / Q000502:physiology; D009419:Nerve Tissue Proteins / Q000235:genetics / Q000378:metabolism; D010300:Parkinson Disease / Q000378:metabolism; D016133:Polymerase Chain Reaction / Q000379:methods; D051843:Synucleins; D013844:Thiazoles / Q000378:metabolism; D014162:Transfection / Q000379:methods; D025801:Ubiquitin / Q000378:metabolism; D051844:alpha-Synuclein; D051846:beta-Synuclein
D000085:Acetates; D007211:Indoles; D009419:Nerve Tissue Proteins; C104584:ONO 3; C497604:SNCA protein, human; C497607:SNCB protein, human; D051843:Synucleins; D013844:Thiazoles; D025801:Ubiquitin; D051844:alpha-Synuclein; D051846:beta-Synuclein; D049452:Green Fluorescent Proteins; C007293:DAPI; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases