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

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.

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Figure 1: Reproducibility of unbiased, global mRNA amplification of mouse cardiomyocytes.
Figure 2: Increased cell-to-cell variation in gene expression among cardiomyocytes from the heart of old as compared with young mice.
Figure 3: H 2 O 2 treatment of MEFs increases cell-to-cell variation in gene expression in parallel to the induction of genome rearrangements.

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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.

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Correspondence to Jan Vijg.

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

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Supplementary Notes

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

Supplementary Table 1

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

Supplementary Figure 1

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

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Bahar, R., Hartmann, C., Rodriguez, K. et al. Increased cell-to-cell variation in gene expression in ageing mouse heart. Nature 441, 1011–1014 (2006). https://doi.org/10.1038/nature04844

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