Parkinson's disease transgenic mitochondrial cybrids generate Lewy inclusion bodies.
Many models of Parkinson's disease (PD) have succeeded in replicating dopaminergic neuron loss or alpha-synuclein aggregation but not the formation of classical Lewy bodies, the pathological hallmark of PD. Our cybrid model of sporadic PD was created by introducing the mitochondrial genes from PD patients into neuroblastoma cells that lack mitochondrial DNA. Previous studies using cybrids have shown that information encoded by mitochondrial DNA in patients contributes to many pathogenic features of sporadic PD. In this paper, we report the generation of fibrillar and vesicular inclusions in a long-term cybrid cell culture model that replicates the essential antigenic and structural features of Lewy bodies in PD brain without the need for exogenous protein expression or inhibition of mitochondrial or proteasomal function. The inclusions generated by PD cybrid cells stained with eosin, thioflavin S, and antibodies to alpha-synuclein, ubiquitin, parkin, synphilin-1, neurofilament, beta-tubulin, the proteasome, nitrotyrosine, and cytochrome c. Future studies of these cybrids will enable us to better understand how Lewy bodies form and what role they play in the pathogenesis of PD.
Trimmer
Patricia A
PA
Center for the Study of Neurodegenerative Diseases and Department of Neurology, University of Virginia, School of Medicine, Charlottesville, Virginia 22908, USA.
Borland
M Kathleen
MK
Keeney
Paula M
PM
Bennett
James P
JP
Parker
W Davis
WD
D000368:Aged; D015153:Blotting, Western; D002352:Carrier Proteins / Q000378:metabolism; D016022:Case-Control Studies; D002460:Cell Line; D003546:Cysteine Endopeptidases / Q000378:metabolism; D045304:Cytochromes c / Q000378:metabolism; D004272:DNA, Mitochondrial / Q000235:genetics* / Q000502:physiology; D042967:Electron Transport Complex I / Q000378:metabolism; D005260:Female; D006801:Humans; D007150:Immunohistochemistry; D016631:Lewy Bodies / Q000235:genetics / Q000378:metabolism* / Q000648:ultrastructure; D008297:Male; D018613:Microscopy, Confocal; D008854:Microscopy, Electron / Q000379:methods; D008875:Middle Aged; D009097:Multienzyme Complexes / Q000378:metabolism; D009419:Nerve Tissue Proteins / Q000378:metabolism; D009447:Neuroblastoma; D016900:Neurofilament Proteins / Q000378:metabolism; D009474:Neurons / Q000378:metabolism / Q000473:pathology*; D010300:Parkinson Disease / Q000235:genetics / Q000378:metabolism*; D011233:Precipitin Tests; D046988:Proteasome Endopeptidase Complex; D013194:Staining and Labeling; D051843:Synucleins; D019076:Transgenes / Q000502:physiology*; D014404:Tubulin / Q000378:metabolism; D014443:Tyrosine / Q000031:analogs & derivatives* / Q000378:metabolism; D025801:Ubiquitin / Q000378:metabolism; D044767:Ubiquitin-Protein Ligases / Q000378:metabolism; D051844:alpha-Synuclein
D002352:Carrier Proteins; D004272:DNA, Mitochondrial; D009097:Multienzyme Complexes; D009419:Nerve Tissue Proteins; D016900:Neurofilament Proteins; C497604:SNCA protein, human; C118958:SNCAIP protein, human; D051843:Synucleins; D014404:Tubulin; D025801:Ubiquitin; D051844:alpha-Synuclein; C002744:3-nitrotyrosine; D014443:Tyrosine; D045304:Cytochromes c; D044767:Ubiquitin-Protein Ligases; C111567:parkin protein; D003546:Cysteine Endopeptidases; D046988:Proteasome Endopeptidase Complex; D042967:Electron Transport Complex I