Abstract
Signaling by the calcium-dependent phosphatase calcineurin profoundly influences the growth and gene expression of cardiac and skeletal muscle. Calcineurin binds to calsarcins, a family of muscle-specific proteins of the sarcomeric Z-disc, a focal point in the pathogenesis of human cardiomyopathies. We show that calsarcin-1 negatively modulates the functions of calcineurin, such that calcineurin signaling was enhanced in striated muscles of mice that do not express calsarcin-1. As a consequence of inappropriate calcineurin activation, mice with a null mutation in calsarcin-1 showed an excess of slow skeletal muscle fibers. The absence of calsarcin-1 also activated a hypertrophic gene program, despite the absence of hypertrophy, and enhanced the cardiac growth response to pressure overload. In contrast, cardiac adaptation to other hypertrophic stimuli, such as chronic catecholamine stimulation or exercise, was not affected. These findings show important roles for calsarcins as modulators of calcineurin signaling and the transmission of a specific subset of stress signals leading to cardiac remodeling in vivo.
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Acknowledgements
We thank A. Tizenor for graphics, J. Page for editorial assistance, and U. Öhl and J. Emma for excellent technical assistance. D. J. Garry provided the running cages for mouse exercise training. The advice of T. Fehm on statistical analyses is appreciated. We are grateful to S. Schiaffino and E. Bush for providing antibodies and H. Weiler (Blood Research Institute) for ES cell targeting. E.N.O. was supported by grants from the US National Institutes of Health, the Donald W. Reynolds Foundation, the Robert A. Welch Foundation and the Texas Advanced Technology Program. T.B was supported by the US National Institutes of Health minority supplement grant. N.F. was supported by grants of the Deutsche Forschungsgemeinschaft (Fr1289/1-1 and Fr1289/3-1) and the Young Investigator Program of the University of Heidelberg.
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Supplementary information
Supplementary Fig. 1
Smaller cardiomyocytes in calsarcin-deficient hearts. (PDF 92 kb)
Supplementary Fig. 2
Calsarcin inhibits calcineurin activity in vitro. (PDF 37 kb)
Supplementary Fig. 3
Calsarcin-1 null/MCIP-transgenic mice display superinduction of ANF-expression. (PDF 97 kb)
Supplementary Fig. 4
Similar degree of exercise-induced hypertrophy in wild-type and calsarcin mutant hearts (PDF 45 kb)
Supplementary Table 1
Transthoracic echocardiographic analysis of wild-type and calsarcin mutant hearts (PDF 112 kb)
Supplementary Table 2
Characterization of left ventricular mechanics by pressure-volume measurements (PDF 143 kb)
Supplementary Table 3
Sequences of primers used in dot-blot and rea-time PCR experiments (PDF 113 kb)
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Frey, N., Barrientos, T., Shelton, J. et al. Mice lacking calsarcin-1 are sensitized to calcineurin signaling and show accelerated cardiomyopathy in response to pathological biomechanical stress. Nat Med 10, 1336–1343 (2004). https://doi.org/10.1038/nm1132
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DOI: https://doi.org/10.1038/nm1132
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