Transmitochondrial embryonic stem cells containing pathogenic mtDNA mutations are compromised in neuronal differentiation

被引:47
作者
Kirby, D. M. [1 ,3 ]
Rennie, K. J. [1 ]
Smulders-Srinivasan, T. K. [1 ]
Acin-Perez, R. [4 ]
Whittington, M. [2 ]
Enriquez, J. -A. [4 ]
Trevelyan, A. J. [1 ]
Turnbull, D. M. [1 ,2 ]
Lightowlers, R. N. [1 ,2 ]
机构
[1] Univ Newcastle, Mitochondrial Res Grp, Sch Med, Inst Ageing & Hlth, Newcastle Upon Tyne NE2 4HH, Tyne & Wear, England
[2] Univ Newcastle, Inst Neurosci, Sch Med, Newcastle Upon Tyne NE2 4HH, Tyne & Wear, England
[3] Royal Childrens Hosp, Murdoch Childrens Res Inst, Melbourne, Vic, Australia
[4] Univ Zaragoza, Biochem & Mol & Cellular Biol Dept, Zaragoza, Spain
基金
澳大利亚国家健康与医学研究理事会; 英国惠康基金;
关键词
MOUSE MITOCHONDRIAL-DNA; MOTOR-NERVE TERMINALS; COMPLEX-I; ES CELLS; DISEASE; MICE; CA2+; RELEASE; INHIBITION; GENERATION;
D O I
10.1111/j.1365-2184.2009.00612.x
中图分类号
Q2 [细胞生物学];
学科分类号
071009 ; 090102 ;
摘要
Objectives: Defects of the mitochondrial genome (mtDNA) cause a series of rare, mainly neurological disorders. In addition, they have been implicated in more common forms of movement disorders, dementia and the ageing process. In order to try to model neuronal dysfunction associated with mitochondrial disease, we have attempted to establish a series of transmitochondrial mouse embryonic stem cells harbouring pathogenic mtDNA mutations. Materials and methods: Transmitochondrial embryonic stem cell cybrids were generated by fusion of cytoplasts carrying a variety of mtDNA mutations, into embryonic stem cells that had been pretreated with rhodamine 6G, to prevent transmission of endogenous mtDNA. Cybrids were differentiated into neurons and assessed for efficiency of differentiation and electrophysiological function. Results: Neuronal differentiation could occur, as indicated by expression of neuronal markers. Differentiation was impaired in embryonic stem cells carrying mtDNA mutations that caused severe biochemical deficiency. Electrophysiological tests showed evidence of synaptic activity in differentiated neurons carrying non-pathogenic mtDNA mutations or in those that caused a mild defect of respiratory activity. Again, however, neurons carrying mtDNA mutations that resulted in severe biochemical deficiency had marked reduction in post-synaptic events. Conclusions: Differentiated neurons carrying severely pathogenic mtDNA defects can provide a useful model for understanding how such mutations can cause neuronal dysfunction.
引用
收藏
页码:413 / 424
页数:12
相关论文
共 33 条
[1]   An intragenic suppressor in the cytochrome c oxidase I gene of mouse mitochondrial DNA [J].
Acín-Pérez, R ;
Bayona-Bafaluy, MP ;
Bueno, M ;
Machicado, C ;
Fernández-Silva, P ;
Pérez-Martos, A ;
Montoya, J ;
López-Pérez, MJ ;
Sancho, J ;
Enríquez, JA .
HUMAN MOLECULAR GENETICS, 2003, 12 (03) :329-339
[2]   The mtDNA-encoded ND6 subunit of mitochondrial NADH dehydrogenase is essential for the assembly of the membrane arm and the respiratory function of the enzyme [J].
Bai, YD ;
Attardi, G .
EMBO JOURNAL, 1998, 17 (16) :4848-4858
[3]   EMBRYONIC STEM-CELLS EXPRESS NEURONAL PROPERTIES IN-VITRO [J].
BAIN, G ;
KITCHENS, D ;
YAO, M ;
HUETTNER, JE ;
GOTTLIEB, DI .
DEVELOPMENTAL BIOLOGY, 1995, 168 (02) :342-357
[4]   Revisiting the mouse mitochondrial DNA sequence [J].
Bayona-Bafaluy, MP ;
Acín-Pérez, R ;
Mullikin, JC ;
Park, JS ;
Moreno-Loshuertos, R ;
Hu, PQ ;
Pérez-Martos, A ;
Fernández-Silva, P ;
Bai, YD ;
Enríquez, JA .
NUCLEIC ACIDS RESEARCH, 2003, 31 (18) :5349-5355
[5]   High levels of mitochondrial DNA deletions in substantia nigra neurons in aging and Parkinson disease [J].
Bender, A ;
Krishnan, KJ ;
Morris, CM ;
Taylor, GA ;
Reeve, AK ;
Perry, RH ;
Jaros, E ;
Hersheson, JS ;
Betts, J ;
Klopstock, T ;
Taylor, RW ;
Turnbull, DM .
NATURE GENETICS, 2006, 38 (05) :515-517
[6]   Inhibition of mitochondrial complex II induces a long-term potentiation of NMDA-mediated synaptic excitation in the striatum requiring endogenous dopamine [J].
Calabresi, P ;
Gubellini, P ;
Picconi, B ;
Centonze, D ;
Pisani, A ;
Bonsi, P ;
Greengard, P ;
Hipskind, RA ;
Borrelli, E ;
Bernardi, G .
JOURNAL OF NEUROSCIENCE, 2001, 21 (14) :5110-5120
[7]   MELAS MUTATION IN MTDNA BINDING-SITE FOR TRANSCRIPTION TERMINATION FACTOR CAUSES DEFECTS IN PROTEIN-SYNTHESIS AND IN RESPIRATION BUT NO CHANGE IN LEVELS OF UPSTREAM AND DOWNSTREAM MATURE TRANSCRIPTS [J].
CHOMYN, A ;
MARTINUZZI, A ;
YONEDA, M ;
DAGA, A ;
HURKO, O ;
JOHNS, D ;
LAI, ST ;
NONAKA, I ;
ANGELINI, C ;
ATTARDI, G .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1992, 89 (10) :4221-4225
[8]   Mitochondrial Ca2+ uptake prevents desynchronization of quantal release and minimizes depletion during repetitive stimulation of mouse motor nerve terminals [J].
David, G ;
Barrett, EF .
JOURNAL OF PHYSIOLOGY-LONDON, 2003, 548 (02) :425-438
[9]   Quantitative estimate of mitochondrial [Ca2+] in stimulated motor nerve terminals [J].
David, G ;
Talbot, J ;
Barrett, EF .
CELL CALCIUM, 2003, 33 (03) :197-206
[10]   A mouse model of mitochondrial disease reveals germline selection against severe mtDNA mutations [J].
Fan, Weiwei ;
Waymire, Katrina G. ;
Narula, Navneet ;
Li, Peng ;
Rocher, Christophe ;
Coskun, Pinar E. ;
Vannan, Mani A. ;
Narula, Jagat ;
MacGregor, Grant R. ;
Wallace, Douglas C. .
SCIENCE, 2008, 319 (5865) :958-962