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The Expression and Role of Protein Kinase C in Neonatal Cardiac Myocyte Attachment, Cell Volume, and Myofibril Formation Is Dependent on the Composition of the Extracellular Matrix

Published online by Cambridge University Press:  12 May 2005

Tara A. Bullard ,
Thomas K. Borg  and
Robert L. Price
Tara A. Bullard
Affiliation:
Center for Cellular and Molecular Cardiology, University of Rochester Medical Center, Rochester, NY 14642, USA
Thomas K. Borg
Affiliation:
Department of Cell and Developmental Biology and Anatomy, University of South Carolina School of Medicine, Columbia, SC 29209, USA
Robert L. Price
Affiliation:
Department of Cell and Developmental Biology and Anatomy, University of South Carolina School of Medicine, Columbia, SC 29209, USA
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Abstract

The extracellular matrix (ECM) is a dynamic component of tissues that influences cellular phenotype and behavior. We sought to determine the role of specific ECM substrates in the regulation of protein kinase C (PKC) isozyme expression and function in cardiac myocyte attachment, cell volume, and myofibril formation. PKC isozyme expression was ECM substrate specific. Increasing concentrations of the PKC δ inhibitor rottlerin attenuated myocyte attachment to randomly organized collagen (1, 5, and 10 μM), laminin (5 and 10 μM), aligned collagen (5 and 10 μM), and fibronectin (10 μM). Rottlerin significantly decreased cell volume on laminin and randomly organized collagen, and inhibited myofibril formation on laminin. The PKC α inhibitor Gö 6976 inhibited attachment to randomly organized collagen at 6 nM but did not affect cell volume. The general PKC inhibitor Bisindolylmalemide I (10 and 30 μM) did not affect myocyte attachment; however, it significantly decreased cell volume on randomly organized collagen. Our data indicate that PKC isozymes are expressed and utilized by neonatal cardiac myocytes during attachment, cell growth, and myofibril formation. Specifically, it appears that PKC δ and/or its downstream effectors play an important role in the interaction between cardiac myocytes and laminin, providing further evidence that the ECM influences cardiac myocyte behavior.

Keywords

signal transductionprotein kinase Cneonatal cardiac myocytesmyofibrilCell Tracker Greenconfocal microscopy
Type
Research Article
Information
Microscopy and Microanalysis , Volume 11 , Issue 3 , June 2005 , pp. 224 - 234
Copyright
© 2005 Microscopy Society of America

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References

REFERENCES

Alt, A., Ohba, M., Li, L., Gartsbein, M., Belanger, A., Denning, M.F., Kuroki, T., Yuspa, S.H., & Tennenbaum, T. (2001). Protein kinase C δ-mediated phosphorylation of α6β4 is associated with reduced integrin localization to the hemidesmosome and decreased keratinocyte attachment. Cancer Res 61, 4591–4598.Google Scholar
Barry, S.T. & Critchley, D.R. (1994). The RhoA-dependent assembly of focal adhesions in Swiss 3T3 cells is associated with increased tyrosine phosphorylation and the recruitment of both pp125FAK and protein kinase C-δ to focal adhesions. J Cell Sci 107, 2033–2045.Google Scholar
Bick, R.J., Snuggs, M.B., Poindexter, B.J., Buja, L.M., & Van Winkle, W.B. (1998). Physical, contractile and calcium handling properties of neonatal cardiac myocytes cultured on different matrices. Cell Adhes Commun 6, 301–310.Google Scholar
Bogoyevitch, M.A., Parker, P.J., & Sugden, P.H. (1993). Characterization of protein kinase C isotype expression in adult rat heart. Protein kinase C-ε is a major isotype present, and it is activated by phorbol esters, epinephrine, and endothelin. Circ Res 72, 757–767.Google Scholar
Borg, T.K., Rubin, K., Lundgren, E., Borg, K., & Obrink, B. (1984). Recognition of extracellular matrix components by neonatal and adult cardiac myocytes. Dev Biol 104, 86–96.Google Scholar
Bouman, R.A., Musters, R.J.P., van Beek-Harmsen, B.J., de Lange, J.J., & Boer, C. (2004). Reactive oxygen species precede protein kinase C-δ activation independent of adenosine triphosphate-sensitive mitochondrial channel opening in sevoflurane-induced cardioprotection. Anesthesiology 100, 506–514.Google Scholar
Bowling, N.B., Walsh, R.A., Song, G., Estridge, T., Sandusky, G.E., Fouts, R.L., Mintze, K., Pickard, T., Roden, R., Bristow, M.R., Sabbah, H.N., Mizrahi, J.L., Gromo, G., King, G.L., & Vlahos, C.J. (1999). Increased protein kinase C activity and expression of Ca2+ sensitive isoforms in the failing human heart. Circulation 99, 384–391.Google Scholar
Burgess, M.L., Buggy, J., Price, R.L., Abel, F.L., Terracio, L., Samarel, A.M., & Borg, T.K. (1996). Exercise- and hypertension-induced collagen changes are related to left ventricular function in rat hearts. Am J Physiol Heart Circ Physiol 270, H151–H159.Google Scholar
Burgess, M.L., McCrea, J.C., & Hedrick, H.L. (2001). Age-associated changes in cardiac matrix and integrins. Mech Ageing Dev 122, 1739–1756.Google Scholar
Carver, W., Nagpal, M.L., Nachtigal, M., Borg, T.K., & Terracio, L. (1991). Collagen expression in mechanically stimulated cardiac fibroblasts. Circ Res 69, 116–122.Google Scholar
Cheng, J.J., Wung, B.S., Chao, Y.J., & Wang, D.L. (2001). Sequential activation of protein kinase C (PKC)-α and PKC-ε contributes to sustained raf/ERK1/2 activation in endothelial cells under mechanical strain. J Biol Chem 276, 313–368.Google Scholar
Davies, S.P., Reddy, H., Caivano, M., & Cohen, P. (2000). Specificity and mechanism of action of some commonly used protein kinase inhibitors. Biochem J 351, 95–105.Google Scholar
Disatnik, M.H., Boutet, S.C., Lee, C.H., Mochly-Rosen, D., & Rando, T.A. (2002). Sequential activation of individual PKC isozymes in integrin-mediated muscle cell spreading: A role for MARCKS in an integrin signaling pathway. J Cell Sci 115, 2151–2163.Google Scholar
Eble, D.M., Straight, J.B., Govindarajan, G., Lou, J., Byron, K.L., & Samarel, A.M. (2000). Endothelin-induced cardiac myocyte hypertrophy: Role for focal adhesion kinase. Am J Physiol Heart Circ Physiol 278, H1696–H1707.Google Scholar
Gschwendt, M., Muller, H.J., Kielbassa, K., Zang, R., Kittstein, W., Rincke, G., & Marks, F. (1994). Rottlerin, a novel protein kinase inhibitor. Biochem Biophys Res Commun 199, 93–98.Google Scholar
Herrmann, K.L., McCulloch, A.D., & Omens, J.H. (2003). Glycated collagen cross-linking alters cardiac mechanics in volume overload hypertrophy. Am J Physiol Heart Circ Physiol 284, H1277–H1284.Google Scholar
Hilenski, L.L., Ma, X.H., Vinson, N., Terracio, L., & Borg, T.K. (1992). The role of β1 integrin in spreading and myofibrillogenesis in neonatal rat cardiomyocytes in vitro. Cell Motil Cytoskeleton 21, 87–100.Google Scholar
Kada, K., Yasui, K., Naruse, K., Kamiya, K., Kodama, I., & Toyama, J. (1999). Orientation change of cardiocytes induced by cyclic stretch stimulation: Time dependency and involvement of protein kinases. J Mol Cell Cardiol 31, 247–259.Google Scholar
Kapadia, R., Yurchenco, P.D., & Amsler, K. (1999). Binding of the renal epithelial cell line LLC-PK1 to laminin is regulated by protein kinase C. J Am Soc Nephrol 10, 1214–1223.Google Scholar
Kumar, A., Chaudhry, I., Reid, M.B., & Boriek, A.M. (2002). Distinct signaling pathways are activated in response to mechanical stress applied axially and transversely to skeletal muscle fibers. J Biol Chem 277, 46493–46503.Google Scholar
Lam, K., Zhang, L., Yamada, K.M., & Lafrenie, R.M. (2001). Adhesion of epithelial cells to fibronectin or collagen I induces alterations in gene expression via a protein kinase C-dependent mechanism. J Cell Physiol 189, 79–90.Google Scholar
Ludwig, L.M., Weihrauch, D., Kersten, J.R., Pagel, P.S., & Warltier, D.C. (2004). Protein kinase C translocation and Src protein tyrosine kinase activation mediate isoflurane-induced preconditioning in vivo. Anesthesiology 100, 532–539.Google Scholar
Martiny-Baron, G., Kazanietz, M.G., Mischak, H., Blumberg, P.M., Kochs, G., Hug, H., Marme, D., & Schachtele, C. (1993). Selective inhibition of protein kinase C isozymes by the indolocarbazole Go 6976. J Biol Chem 268, 9194–9197.Google Scholar
Milner, R. & Campbell, I.L. (2002). Cytokines regulate microglial adhesion to laminin and astrocyte extracellular matrix via protein kinase C-dependent activation of the α6β1 integrin. J Neurosci 22, 1562–1572.Google Scholar
Oh, C.-D., Kang, S.S., Ha, M.J., & Chun, J.S. (1999). Analysis of protein kinase C isoforms involved in the activation of laminin receptor in Raw264.7 macrophages. IUBMB Life 48, 439–443.Google Scholar
Owen, P.J., Johnson, G.D., & Lord, J.M. (1996). Protein kinase C-δ associates with vimentin intermediate filaments in differentiated HL60 cells. Exp Cell Res 225, 366–373.Google Scholar
Parker, P.J. (1999). Inhibition of protein kinase C—Do we, can we and should we? Pharmacol Ther 82, 263–267.Google Scholar
Ron, D. & Kazanietz, M.G. (1999). New insights into the regulation of protein kinase C and novel phorbol ester receptors. FASEB J 13, 1658–1676.Google Scholar
Ross, R.S. & Borg, T.K. (2001). Integrins and the myocardium. Circ Res 88, 1112–1119.Google Scholar
Rybin, V.O. & Steinberg, S.F. (1994). Protein kinase C isoform expression and regulation in the developing rat heart. Circ Res 74, 299–309.Google Scholar
Simpson, D.G., Terracio, L., Terracio, M., Price, R.L., Turner, D.C., & Borg, T.K. (1994). Modulation of cardiac myocyte phenotype in vitro by the composition and orientation of the extracellular matrix. J Cell Physiol 161, 89–105.Google Scholar
Steinberg, S.F., Goldberg, M., & Rybin, V.O. (1995). Protein kinase C isoform diversity in the heart. J Mol Cell Cardiol 27, 141–153.Google Scholar
Toullec, D., Pianetti, P., Coste, H., Bellevergue, P., Grand-Perret, T., Ajakane, M., Baudet, V., Boissin, P., Boursier, E., Loriolle, F., Duhamel, L., Charon, D., & Kirilovsky, J. (1991). The Bisindolylmaleimide GF 109203X is a potent and selective inhibitor of protein kinase C. J Biol Chem 266, 15771–15781.Google Scholar
Watters, D., Garrone, B., Gobert, G., Williams, S., Gardiner, R., & Lavin, M. (1996). Bistratene A causes phosphorylation of talin and redistribution of actin microfilaments in fibroblasts: Possible role for PKC-δ. Exp Cell Res 229, 327–335.Google Scholar
Woods, A. & Couchman, J.R. (1992). Protein kinase C involvement in focal adhesion formation. J Cell Sci 101, 277–290.Google Scholar