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Differences in reactive oxygen species production explain the phenotypes associated with common mouse mitochondrial DNA variants

Nature Genetics volume 38, pages 1261–1268 (2006)Cite this article

Abstract

Common mitochondrial DNA (mtDNA) haplotypes in humans and mice have been associated with various phenotypes, including learning performance and disease penetrance. Notably, no influence of mtDNA haplotype in cell respiration has been demonstrated. Here, using cell lines carrying four different common mouse mtDNA haplotypes in an identical nuclear background, we show that the similar level of respiration among the cell lines is only apparent and is a consequence of compensatory mechanisms triggered by different production of reactive oxygen species. We observe that the respiration capacity per molecule of mtDNA in cells with the NIH3T3 or NZB mtDNA is lower than in those with the C57BL/6J, CBA/J or BALB/cJ mtDNA. In addition, we have determined the genetic element underlying these differences. Our data provide insight into the molecular basis of the complex phenotypes associated with common mtDNA variants and anticipate a relevant contribution of mtDNA single nucleotide polymorphisms to phenotypic variability in humans.

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Figure 1: Respiration capacity and relative growth of transmitochondrial cybrids in galactose and glucose medium.
Figure 2: ROS, defense activity and mtDNA amount in transmitochondrial cybrids with different mtDNA haplotypes.
Figure 3: Adaptation to galactose of transmitochondrial cybrids with different mtDNA haplotypes.
Figure 4: Activity of the Krebs cycle enzymes is influenced by mtDNA haplotype.

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Acknowledgements

We thank C. Moraes and E. Shoubridge for the NZB hepatocytes; E. Ruiz-Pesini, A. Barrientos, C. López-Otin and M. Palacín for comments on the manuscript; and S. Morales for technical assistance. Our work was supported by the Spanish Ministry of Education (SAF2003-00103), the Instituto de Salud Carlos III (REDEMETH-G03/054, REDCIEN C03/06-Grupo RC-N34-3 yr ECEMECRE G03/011), the European Union (EUMITOCOMBAT-LSHM-CT-2004-503116), Group of Excellence grant DGA (B55) and Fundación Ramón Areces. R.M.-L. and R.A.-P. are supported by a predoctoral fellowship from the Spanish Ministry of Education and M.E.G. is supported by an I3P postdoctoral contract from the CSIC, Spain.

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Authors and Affiliations

  1. Departamento de Bioquímica y Biología Molecular y Celular, Universidad de Zaragoza, Miguel Servet 177, Zaragoza, 50013, Spain

    Raquel Moreno-Loshuertos, Rebeca Acín-Pérez, Patricio Fernández-Silva, Nieves Movilla, Acisclo Pérez-Martos & José Antonio Enríquez

  2. Departamento de Inmunología, Centro de Investigaciones Biológicas, Consejo Superior de Investigaciones Científicas (CSIC), Avda Ramiro de Maeztu 9, Madrid, 28040, Spain

    Santiago Rodriguez de Cordoba & M Esther Gallardo

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Contributions

This study was designed and the paper was written by P.F.-S., A.P.-M., S.R.d.C. and J.A.E.; the wild-type cybrid cell line was constructed and phenotype assessed by R.M.-L. and R.A.-P.; the TmND6ko mtDNA mutant cell line was generated and characterized by N.M.; and the mtDNA resequencing strategy was designed by, and full mtDNA sequencing performed by, M.E.G.

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Correspondence to José Antonio Enríquez.

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The authors declare no competing financial interests.

Supplementary information

Supplementary Fig. 1

RFLP analysis to detect the possible presence of L929 mtDNA in the transmitochondrial cell lines. (PDF 112 kb)

Supplementary Fig. 2

Protein levels for specific subunits of the mtETC complexes in the different mtDNA haplotypes. (PDF 104 kb)

Supplementary Fig. 3

ND3/ND4 relative transcript levels in the transmitochondrial cell lines. (PDF 98 kb)

Supplementary Table 1

mtDNA nucleotide differences between C57BL/6J and NZB/B1NJ. (PDF 18 kb)

Supplementary Table 2

Primer sequences. (PDF 80 kb)

Supplementary Methods (PDF 96 kb)

Supplementary Note (PDF 24 kb)

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Moreno-Loshuertos, R., Acín-Pérez, R., Fernández-Silva, P. et al. Differences in reactive oxygen species production explain the phenotypes associated with common mouse mitochondrial DNA variants. Nat Genet 38, 1261–1268 (2006). https://doi.org/10.1038/ng1897

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