Abstract
Cardiac hypertrophy is a compensatory response to a variety of physiologi cal or pathological stimuli. However, prolonged hypertrophic responses may eventually lead to heart failure, arrhythmia, and sudden death. A number of intracellular signaling pathways have been implicated to play a critical role in the regulation of cardiac hypertrophy. In this chapter, the mitogen-activated protein kinase signaling pathway is used to illustrate conventional assays to detect the expression, phosphorylation, and activation of signaling proteins during cardiac hypertrophy, including Western blot, immunohistochemical staining, and immune complex kinase assays. Newly emerging techniques for analyzing cell signaling are also discussed in this chapter. Identifying and char acterizing the expression and activation of these signaling proteins will pro vide important insights into the mechanisms that regulate hypertrophic cell growth and assist in development of new therapeutic approaches to limit car diac hypertrophy.
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References
Molkentin, J. D. and Dorn, I. G. 2nd. (2001) Cytoplasmic signaling pathways that regulate cardiac hypertrophy. Annu. Rev. Physiol. 63, 391–426.
Aikawa, R., Komuro, I., Yamazaki, T., et al. (1999) Rho family small G proteins play critical roles in mechanical stress-induced hypertrophic responses in cardiac myocytes. Circ. Res. 84, 458–466.
Clerk, A. and Sugden, P. H. (2000) Small guanine nucleotide-binding proteins and myocardial hypertrophy. Circ. Res. 86, 1019–1023.
Sugden, P. H. (1999) Signaling in myocardial hypertrophy: life after calcineurin? Circ. Res. 84, 633–646.
Williams, R. S. (2002) Calcineurin signaling in human cardiac hypertrophy. Cir culation 105, 2242–2243.
Bueno, O. F., De Windt, L. J., Tymitz, K. M., et al. (2000) The MEK1-ERK1/2 signaling pathway promotes compensated cardiac hypertrophy in transgenic mice. EMBO J. 19, 6341–6350.
Bueno, O. F. and Molkentin, J. D. (2002) Involvement of extracellular signal-regu lated kinases 1/2 in cardiac hypertrophy and cell death. Circ. Res. 91, 776–781.
Choukroun, G., Hajjar, R., Fry, S., et al. (1999) Regulation of cardiac hypertro phy in vivo by the stress-activated protein kinases/c-Jun NH(2)-terminal kinases. J. Clin. Invest. 104, 391–398.
Haq, S., Choukroun, G., Lim, H., et al. (2001) Differential activation of signal transduction pathways in human hearts with hypertrophy versus advanced heart failure. Circulation 103, 670–677.
Takeishi, Y., Huang, Q., Abe, J., et al. (2001) Src and multiple MAP kinase acti vation in cardiac hypertrophy and congestive heart failure under chronic pressure-overload: comparison with acute mechanical stretch. J. Mol. Cell. Cardiol. 33, 1637–1648.
Takeishi, Y., Ping, P., Bolli, R., Kirkpatrick, D. L., Hoit, B. D., and Walsh, R. A. (2000) Transgenic overexpression of constitutively active protein kinase C epsi lon causes concentric cardiac hypertrophy. Circ. Res. 86, 1218–1223.
Haq, S., Choukroun, G., Kang, Z. B., et al. (2000) Glycogen synthase kinase-3beta is a negative regulator of cardiomyocyte hypertrophy. J. Cell. Biol. 151, 117–130.
Hardt, S. E. and Sadoshima, J. (2002) Glycogen synthase kinase-3beta: a novel regulator of cardiac hypertrophy and development. Circ. Res. 90, 1055–1063.
Booz, G. W., Day, J. N., and Baker, K. M. (2002) Interplay between the cardiac renin angiotensin system and JAK-STAT signaling: role in cardiac hypertrophy, ischemia/reperfusion dysfunction, and heart failure. J. Mol. Cell. Cardiol. 34, 1443–1453.
Hirotani, S., Otsu, K., Nishida, K., et al. (2002) Involvement of nuclear factor-kappaB and apoptosis signal-regulating kinase 1 in G-protein-coupled receptor agonist-induced cardiomyocyte hypertrophy. Circulation 105, 509–515.
Lange-Carter, C. A., Pleiman, C. M., Gardner, A. M., Blumer, K. J., and Johnson, G. L. (1993) A divergence in the MAP kinase regulatory network defined by MEK kinase and Raf. Science 260, 315–319.
Sugden, P. H. and Clerk, A. (1998) “Stress-responsive” mitogen-activated protein kinases (c-Jun N-terminal kinases and p38 mitogen-activated protein kinases) in the myocardium. Circ. Res. 83, 345–352.
Lee, J. D., Ulevitch, R. J., and Han, J. (1995) Primary structure of BMK1: a new mammalian map kinase. Biochem. Biophys. Res. Commun. 213, 715–724.
Widmann, C., Gibson, S., Jarpe, M. B., and Johnson, G. L. (1999) Mitogen-acti vated protein kinase: conservation of a three-kinase module from yeast to human. Physiol. Rev. 79, 143–180.
Rapacciuolo, A., Esposito, G., Caron, K., Mao, L., Thomas, S. A., and Rockman, H. A. (2001) Important role of endogenous norepinephrine and epinephrine in the development of in vivo pressure-overload cardiac hypertrophy. J. Am. Coll. Cardiol. 38, 876–882.
Gillespie-Brown, J., Fuller, S. J., Bogoyevitch, M. A., Cowley, S., and Sugden, P. H. (1995) The mitogen-activated protein kinase kinase MEK1 stimulates a pat tern of gene expression typical of the hypertrophic phenotype in rat ventricular cardiomyocytes. J. Biol. Chem. 270, 28,092–28,096.
Kodama, H., Fukuda, K., Pan, J., et al. (2000) Significance of ERK cascade com pared with JAK/STAT and PI3-K pathway in gp130-mediated cardiac hypertro phy. Am. J. Physiol. Heart Circ. Physiol. 279, H1635–1644.
Yue, T. L., Gu, J. L., Wang, C., et al. (2000) Extracellular signal-regulated kinase plays an essential role in hypertrophic agonists, endothelin-1 and phenylephrine-induced cardiomyocyte hypertrophy. J. Biol. Chem. 275, 37,895–37,901.
Clerk, A., Michael, A., and Sugden, P. H. (1998) Stimulation of the p38 mitogen activated protein kinase pathway in neonatal rat ventricular myocytes by the G protein-coupled receptor agonists, endothelin-1 and phenylephrine: a role in car diac myocyte hypertrophy? J. Cell. Biol. 142, 523–535.
Wang, Y., Huang, S., Sah, V. P., et al. (1998) Cardiac muscle cell hypertrophy and apoptosis induced by distinct members of the p38 mitogen-activated protein kinase family. J. Biol. Chem. 273, 2161–2168.
Nemoto, S., Sheng, Z., and Lin, A. (1998) Opposing effects of Jun kinase and p38 mitogen-activated protein kinases on cardiomyocyte hypertrophy. Mol. Cell. Biol. 18, 3518–3526.
Komuro, I., Kudo, S., Yamazaki, T., Zou, Y., Shiojima, I., and Yazaki, Y. (1996) Mechanical stretch activates the stress-activated protein kinases in cardiac myocytes. FASEB J. 10, 631–636.
Thorburn, J., Xu, S., and Thorburn, A. (1997) MAP kinase-and Rho-dependent signals interact to regulate gene expression but not actin morphology in cardiac muscle cells. EMBO J. 16, 1888–1900.
Ramirez, M. T., Sah, V. P., Zhao, X. L., Hunter, J. J., Chien, K. R., and Brown, J. H. (1997) The MEKK-JNK pathway is stimulated by alpha1-adrenergic receptor and ras activation and is associated with in vitro and in vivo cardiac hypertrophy. J. Biol. Chem. 272, 14,057–14,061.
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Si, X., Rahmani, M., Yuan, J., Luo, H. (2005). Detection of Cardiac Signaling in the Injured and Hypertrophied Heart. In: Sun, Z. (eds) Molecular Cardiology. Methods in Molecular Medicine™, vol 112. Humana Press. https://doi.org/10.1007/978-1-59259-879-3_19
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DOI: https://doi.org/10.1007/978-1-59259-879-3_19
Publisher Name: Humana Press
Print ISBN: 978-1-58829-363-3
Online ISBN: 978-1-59259-879-3
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