title: Cybrid models of Parkinson's disease show variable mitochondrial biogenesis and genotype-respiration relationships.
abstract: Sporadic Parkinson's disease (sPD) is a nervous system-wide disease that presents with a bradykinetic movement disorder and frequently progresses to include depression and cognitive impairment. Cybrid models of sPD are based on expression of sPD platelet mitochondrial DNA (mtDNA) in neural cells and demonstrate some similarities to sPD brains. In sPD and CTL cybrids we characterized aspects of mitochondrial biogenesis, mtDNA genomics, composition of the respirasome and the relationships among isolated mitochondrial and intact cell respiration. Cybrid mtDNA levels varied and correlated with expression of PGC-1 alpha, a transcriptional co-activator regulator of mitochondrial biogenesis. Levels of mtDNA heteroplasmic mutations were asymmetrically distributed across the mitochondrial genome; numbers of heteroplasmies were more evenly distributed. Neither levels nor numbers of heteroplasmies distinguished sPD from CTL. sPD cybrid mitochondrial ETC subunit protein levels were not altered. Isolated mitochondrial complex I respiration rates showed limited correlation with whole cell complex I respiration rates in both sPD and CTL cybrids. Intact cell respiration during the normoxic-anoxic transition yielded K(m) values for oxygen that directly related to respiration rates in CTL but not in sPD cell lines. Both sPD and CTL cybrid cells are substantially heterogeneous in mitochondrial genomic and physiologic properties. Our results suggest that mtDNA depletion may occur in sPD neurons and could reflect impairment of mitochondrial biogenesis. Cybrids remain a valuable model for some aspects of sPD but their heterogeneity mitigates against a simple designation of sPD phenotype in this cell model.
authors.affiliation: Morris K. Udall Parkinson's Research Center of Excellence, University of Virginia, PO Box 800394, Charlottesville, VA 22908, USA.
authors.forename: Paula M
authors.initials: PM
authors.lastname: Keeney
authors.forename: Lisa D
authors.initials: LD
authors.lastname: Dunham
authors.forename: Caitlin K
authors.initials: CK
authors.lastname: Quigley
authors.forename: Stephanie L
authors.initials: SL
authors.lastname: Morton
authors.forename: Kristen E
authors.initials: KE
authors.lastname: Bergquist
authors.forename: James P
authors.initials: JP
authors.lastname: Bennett
mesh_terms: D000368:Aged; D020137:Base Pair Mismatch; D002460:Cell Line; D003470:Culture Media; D017931:DNA Primers; D004272:DNA, Mitochondrial / Q000096:biosynthesis / Q000235:genetics; D005260:Female; D018628:Gene Dosage; D005838:Genotype; D006801:Humans; D006822:Hybrid Cells; D016014:Linear Models; D008297:Male; D008875:Middle Aged; D008928:Mitochondria / Q000378:metabolism* / Q000473:pathology; D010101:Oxygen Consumption / Q000235:genetics / Q000502:physiology*; D010300:Parkinson Disease / Q000235:genetics / Q000473:pathology / Q000503:physiopathology*; D020133:Reverse Transcriptase Polymerase Chain Reaction
chemical_list: D003470:Culture Media; D017931:DNA Primers; D004272:DNA, Mitochondrial