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Am J Physiol Heart Circ Physiol 280: H756-H766, 2001;
0363-6135/01 $5.00
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Vol. 280, Issue 2, H756-H766, February 2001

Role of protein kinase C-epsilon in hypertrophy of cultured neonatal rat ventricular myocytes

James B. Strait III1, Jody L. Martin1, Allison Bayer1, Ruben Mestril1, Diane M. Eble2, and Allen M. Samarel1,2

The Cardiovascular Institute and the Departments of 1 Physiology and 2 Medicine, Loyola University Chicago Stritch School of Medicine, Maywood, Illinois 60153

Using adenovirus (Adv)-mediated overexpression of constitutively active (ca) and dominant-negative (dn) mutants, we examined whether protein kinase C (PKC)-epsilon , the major novel PKC isoenzyme expressed in the adult heart, was necessary and/or sufficient to induce specific aspects of the hypertrophic phenotype in low-density, neonatal rat ventricular myocytes (NRVM) in serum-free culture. Adv-caPKC-epsilon did not increase cell surface area or the total protein-to-DNA ratio. However, cell shape was markedly affected, as evidenced by a 67% increase in the cell length-to-width ratio and a 17% increase in the perimeter-to-area ratio. Adv-caPKC-epsilon also increased atrial natriuretic factor (ANF) and beta -myosin heavy chain (MHC) mRNA levels 2.5 ± 0.3- and 2.1 ± 0.2-fold, respectively, compared with NRVM infected with an empty, parent vector (P < 0.05 for both). Conversely, Adv-dnPKC-epsilon did not block endothelin-induced increases in cell surface area, the total protein-to-DNA ratio, or upregulation of beta -MHC and ANF gene expression. However, the dominant-negative inhibitor markedly suppressed endothelin-induced extracellular signal-regulated kinase (ERK) 1/2 activation. Taken together, these results indicate that caPKC-epsilon overexpression alters cell geometry, producing cellular elongation and remodeling without a significant, overall increase in cell surface area or total protein accumulation. Furthermore, PKC-epsilon activation and downstream signaling via the ERK cascade may not be necessary for cell growth, protein accumulation, and gene expression changes induced by endothelin.

endothelin-1; extracellular signal-regulated kinase; signal transduction; heart; adenovirus


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