Letter | Published:

Increased cell-to-cell variation in gene expression in ageing mouse heart

Naturevolume 441pages10111014 (2006) | Download Citation

Subjects

Abstract

The accumulation of somatic DNA damage has been implicated as a cause of ageing in metazoa1,2. One possible mechanism by which increased DNA damage could lead to cellular degeneration and death is by stochastic deregulation of gene expression. Here we directly test for increased transcriptional noise in aged tissue by dissociating single cardiomyocytes from fresh heart samples of both young and old mice, followed by global mRNA amplification and quantification of mRNA levels in a panel of housekeeping and heart-specific genes. Although gene expression levels already varied among cardiomyocytes from young heart, this heterogeneity was significantly elevated at old age. We had demonstrated previously an increased load of genome rearrangements and other mutations in the heart of aged mice3,4. To confirm that increased stochasticity of gene expression could be a result of increased genome damage, we treated mouse embryonic fibroblasts in culture with hydrogen peroxide. Such treatment resulted in a significant increase in cell-to-cell variation in gene expression, which was found to parallel the induction and persistence of genome rearrangement mutations at a lacZ reporter locus. These results underscore the stochastic nature of the ageing process, and could provide a mechanism for age-related cellular degeneration and death in tissues of multicellular organisms.

Access optionsAccess options

Rent or Buy article

Get time limited or full article access on ReadCube.

from$8.99

All prices are NET prices.

References

  1. 1

    Vijg, J. & Dollé, M. E. Large genome rearrangements as a primary cause of aging. Mech. Ageing Dev. 123, 907–915 (2002)

  2. 2

    Kirkwood, T. B. Understanding the odd science of aging. Cell 120, 437–447 (2005)

  3. 3

    Dollé, M. E., Snyder, W. K., Gossen, J. A., Lohman, P. H. & Vijg, J. Distinct spectra of somatic mutations accumulated with age in mouse heart and small intestine. Proc. Natl Acad. Sci. USA 97, 8403–8408 (2000)

  4. 4

    Dollé, M. E. & Vijg, J. Genome dynamics in aging mice. Genome Res. 12, 1732–1738 (2002)

  5. 5

    Levsky, J. M., Shenoy, S. M., Pezo, R. C. & Singer, R. H. Single-cell gene expression profiling. Science 297, 836–840 (2002)

  6. 6

    Levsky, J. M. & Singer, R. H. Gene expression and the myth of the average cell. Trends Cell Biol. 13, 4–6 (2003)

  7. 7

    Klein, C. A. et al. Combined transcriptome and genome analysis of single micrometastatic cells. Nature Biotechnol. 20, 387–392 (2002)

  8. 8

    Kang, P. M., Haunstetter, A., Aoki, H., Usheva, A. & Izumo, S. Morphological and molecular characterization of adult cardiomyocyte apoptosis during hypoxia and reoxygenation. Circ. Res. 87, 118–125 (2000)

  9. 9

    Kaern, M., Elston, T. C., Blake, W. J. & Collins, J. J. Stochasticity in gene expression: from theories to phenotypes. Nature Rev. Genet. 6, 451–464 (2005)

  10. 10

    Becskei, A., Kaufmann, B. B. & van Oudenaarden, A. Contributions of low molecule number and chromosomal positioning to stochastic gene expression. Nature Genet. 37, 937–944 (2005)

  11. 11

    Parrinello, S. et al. Oxygen sensitivity severely limits the replicative lifespan of murine fibroblasts. Nature Cell Biol. 5, 741–747 (2003)

  12. 12

    Boerrigter, M. E., Dollé, M. E., Martus, H. J., Gossen, J. A. & Vijg, J. Plasmid-based transgenic mouse model for studying in vivo mutations. Nature 377, 657–659 (1995)

  13. 13

    Orgel, L. E. The maintenance of the accuracy of protein synthesis and its relevance to ageing. Proc. Natl Acad. Sci. USA 49, 517–521 (1963)

  14. 14

    Finch, C. E. & Kirkwood, T. B. Chance, Development, and Aging (Oxford Univ. Press, New York, 2000)

  15. 15

    Rea, S. L., Wu, D., Cypser, J. R., Vaupel, J. W. & Johnson, T. E. A stress-sensitive reporter predicts longevity in isogenic populations of Caenorhabditis elegans. Nature Genet. 37, 894–898 (2005)

  16. 16

    Golden, T. R. & Melov, S. Microarray analysis of gene expression with age in individual nematodes. Aging Cell 3, 111–124 (2004)

  17. 17

    Fraser, H. B., Hirsh, A. E., Giaever, G., Kumm, J. & Eisen, M. B. Noise minimization in eukaryotic gene expression. PLoS Biol. 2, e137 (2004)

  18. 18

    Busuttil, R. A., Rubio, M., Dollé, M. E., Campisi, J. & Vijg, J. Oxygen accelerates the accumulation of mutations during the senescence and immortalization of murine cells in culture. Aging Cell 2, 287–294 (2003)

  19. 19

    Dollé, M. E., Martus, H. J., Gossen, J. A., Boerrigter, M. E. & Vijg, J. Evaluation of a plasmid-based transgenic mouse model for detecting in vivo mutations. Mutagenesis 11, 111–118 (1996)

Download references

Acknowledgements

This work was supported by an NIH grant to J.V. and a BioFuture grant from the German Federal Ministry for Education and Science to C.A.K. We thank K. Khrapko and R. Beems for advice and useful suggestions. Author Contributions J.V. conceived, designed and supervised the study (initially with M.E.T.D.), obtained the funding and took the primary role in writing the paper. R.B. designed and performed the experiments, assisted by K.A.R., A.D.D. and R.A.B. R.B.C., G.B.C. and B.H.P. performed statistical analyses and made the figures. C.H.H. and C.A.K. provided the modified protocol for global mRNA amplification.

Author information

Affiliations

  1. Buck Institute for Age Research, Novato, California, 94945, USA

    • Rumana Bahar
    • , Rita A. Busuttil
    • , R. Brent Calder
    •  & Jan Vijg
  2. Institut für Immunologie, Ludwig-Maximilians Universität, Germany, 80336, München

    • Claudia H. Hartmann
    •  & Christoph A. Klein
  3. University of Texas Health Science Center, San Antonio, Texas, 78245, USA

    • Karl A. Rodriguez
    • , Ashley D. Denny
    • , Gary B. Chisholm
    •  & Brad H. Pollock
  4. National Institute of Public Health and the Environment, Bilthoven, 3720 BA, The Netherlands

    • Martijn E. T. Dollé

Authors

  1. Search for Rumana Bahar in:

  2. Search for Claudia H. Hartmann in:

  3. Search for Karl A. Rodriguez in:

  4. Search for Ashley D. Denny in:

  5. Search for Rita A. Busuttil in:

  6. Search for Martijn E. T. Dollé in:

  7. Search for R. Brent Calder in:

  8. Search for Gary B. Chisholm in:

  9. Search for Brad H. Pollock in:

  10. Search for Christoph A. Klein in:

  11. Search for Jan Vijg in:

Competing interests

Reprints and permissions information is available at npg.nature.com/reprintsandpermissions. The authors declare no competing financial interests.

Corresponding author

Correspondence to Jan Vijg.

Supplementary information

  1. Supplementary Notes

    This file contains Supplementary Methods and Supplementary Figure 1 Legend. (DOC 41 kb)

  2. Supplementary Table 1

    Selected genes for real-time PCR quantitative analysis in single cells (DOC 106 kb)

  3. Supplementary Figure 1

    Possible effects of enzymatic dissociation or age of the animal on gene expression in cardiac tissues. (PDF 16 kb)

About this article

Publication history

Received

Accepted

Issue Date

DOI

https://doi.org/10.1038/nature04844

Further reading

Comments

By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.