Summary
Norepinephrine stimulates the growth in size of nondividing neonatal cardiocytes. During this time the neonatal cardiocyte is in a period of transition in which the cell can synthesize DNA and yet does not divide. Because the cell undergoes karyokinesis without cytokinesis the objective of this study was to determine whether the norepinephrine-induced growth in size of the neonatal cardiocyte was accompanied by an increase in a) the number of cardiocytes synthesizing DNA, b) the number of binucleate cardiocytes, and c) organized myofibrils. One- to four-d-old neonatal rat heart cells were isolated and placed in serum-free medium which was then supplemented with serum, norepinephrine, norepinephrine plus propranolol, or isoproterenol. After 4 d the number and size of the cells was determined using a Coulter counter. In other cultures cardiocytes were fixed on Days 0, 1, 2, and 4, and an increase in the number of binucleate cardiocytes was found in all treatment groups including controls. However, the rate of binucleation was faster in the norepinephrine group. It was also determined by proliferating cell nuclear antigen (PCNA) antibody staining that by Day 4, over 50% of the cardiocytes were in the cell cycle. The percentage of cells in which PCNA could be detected was higher in the norepinephrine and norepinephrine plus propranolol groups. Furthermore, there was a concomitant increase in the amount and organization of myofibrils in the catecholamine-treated cardiocytes.
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Anversa, P.; Olivetti, G.; Loud A. V. Morphometric study of early postnatal development in the left and right ventricular myocardium of the rat. I. Hypertrophy, hyperplasia and binucleation of myocytes. Cir. Res. 46:495–502; 1980.
Baserga, R. Introduction to cell growth: growth in size and DNA replication. Handb. Exp. Pharmacol. 57:1–12; 1981.
Bishopric, N. H.; Simpson, P. C.; Ordahl, C. P. Induction of the skeletal α-actin gene in α1 hypertrophy of rat cardiac myocytes. J. Clin. Invest. 80:1194–1199; 1987.
Blaes, N.; Boissel, J. Growth-stimulating effect of catecholamines on rat aortic smooth muscle cells in culture. J. Cell Physiol. 116:167–172; 1988.
Bravo, R.; Macdonald-Bravo, H. Existence of two populations of cyclin/proliferating cell nuclear antigen during the cell cycle: association with DNA replication sites. J. Cell Biol. 105:1549–1554; 1987.
Bravo, R.; Blundell, P. A.; Macdonald-Bravo, H. Cyclin/PCNA is the auxiliary protein of DNA polymerase-γ. Nature 326:515–517; 1987.
Brodsky, W. Y.; Arefyeva, A. M.; Uryvaeva I. V. Mitotic polyploidization of mouse heart myocytes during the first postnatal week. Cell Tissue Res. 210:133–144; 1980.
Bugaisky, L.; Zak, R. Biological mechanisms of hypertrophy. In: Fozzard, H. A.; Haber, E.; Jennings, R. B., et al. eds. The heart and cardiovascular system. New York; Raven Press: 1986:1491–1506.
Clubb, F. J.; Bishop, S. P. Formation of binucleated myocardial cells in the neonatal rat. An index for growth hypertrophy. Lab. Invest. 50:571–577; 1984.
Connolly, J. A.; Sarabia, V. E.; Kelvin, D. J., et al. The disappearance of a cyclin-like protein and the appearance of statin is correlated with the onset of differentiation during myogenesis in vitro. Exp. Cell. Res. 174:461–471; 1988.
Cooper, G.; Mercer, W. E.; Hoober, J. K., et al. Load regulation of the properties of adult feline cardiocytes. The role of substrate adhesion. Circ. Res. 58:692–705; 1986.
Cruise, J. L.; Michalopoulos, G. Norepinephrine and epidermal growth factor: dynamics of their interaction in the stimulation of hepatocyte DNA synthesis. J. Cell. Physiol. 125:45–50; 1985.
Cruise, J. L.; Houck, K. A.; Michalopoulos, G. K. Induction of DNA synthesis in cultured rat hepatocytes through stimulation of α1 by norepinephrine. Science 227:749–751; 1985.
Gerdes, A. M.; Moore, J. A.; Hines, J. M., et al. Regional differences in myocyte size in normal rat heart. Anat. Rec. 215:420–426; 1986.
Jaskulski, D.; Gatti, C.; Travali, S.; et al. Regulation of the proliferating cell nuclear antigen cyclin and thymidine kinase mRNA levels by growth factors. J. Biol. Chem. 263:10174–10179; 1988.
Marino, T. A.; Kuseryk, L.; Lauva, I. K. Role of contraction in the structure and growth of neonatal rat cardiocytes. Am. J. Physiol. 253:H1391–1399; 1987.
Marino, T. A.; Kuseryk, L. Factors regulating neonatal cardiocyte growth in size and myofibrils. Anat. Rec. 218:88A; 1987.
Marino, T. A.; Walter, R. A.; D'Ambra, K., et al. The effects of catecholamines on fetal, rat cardiocytes in vitro. Am. J. Anat. 186:127–132; 1989.
Meidell, R. S.; Sen, A.; Henderson, S. A., et al. α1 stimulation of rat myocardial cells increases protein synthesis. Am. J. Physiol. 251:H1076-H1084; 1986.
Olivetti, G.; Anversa, P.; Loud, A. V. Morphometric study of early postnatal development in left and right ventricular myocardium of the rat II. Tissue composition, capillary growth and sarcoplasmic alterations. Circ. Res. 46:503–512; 1980.
Prelich, G.; Kostura, M.; Marchak, D. R. et al. The cell-cycle regulated proliferating cell nuclear antigen is required for SV40 DNA replication in vitro. Nature 326:471–475; 1987a.
Prelich, G.; Tan, C.; Kostura, M., et al. Functional identity of proliferating cell nuclear antigen and a DNA polymerase-γ auxiliary protein. Nature 326:517–520; 1987.
Sherline, P.; Mascardo, R. Catecholamines are mitogenic in 3T3 and bovine aortic endothelial cells. J. Clin. Invest. 74:483–487; 1984.
Simpson, P. Norepinephrine-stimulated hypertrophy of cultured rat myocardial cells is an alpha, adrenergic response. J. Clin. Invest. 72:732–738; 1983.
Simpson, P. Stimulation of hypertrophy of cultured neonatal rat heart cells through an α1 receptor and induction of beating through and α1 and α1 receptor interaction. Evidence for independent regulation of growth and beating. Circ. Res. 56:884–894; 1985.
Simpson, P.N.; Bishopric, N. S.; Coughlin, J. et al. Dual trophic effects of the alpha1-adrenergic receptor in cultured neonatal heart muscle cells. J. Mol. Cell. Cardiol. 18(Supplement 5): 45–58; 1986.
Simpson, P.; McGrath, A.; Savion, S. Myocyte hypertrophy in neonatal rat heart cultures and its regulation by serum and by catecholamines. Circ. Res. 51:787–801; 1982.
Starksen, N. F.; Simpson, P. C.; Bishopric, N. et al. Cardiac myocyte hypertrophy is associated with c-myc protooncogene expression. Proc. Natl. Acad. Sci. USA 83:8349–8350; 1986.
Tan, C.; Castillo, C.; So, A. G., et al. An auxiliary protein for DNA polymerase-γ from fetal calf thymus. J. Biol. Chem. 26:12310–12316; 1986.
Ueno, H. Perryman, M. B.; Roberts, R., et al. Differentiation of cardiac myocytes after mitogen withdrawal exhibits three sequential states of the ventricular growth response. J. Cell Biol. 107:1911–1918; 1988.
Zak, R. Factors controlling cardiac growth. In: Zak, R., ed Growth of the heart in health and disease. New York: Raven Press; 1984:165–185.
Zuber, M.; Tan, E. M.; Ryoji, M. Involvement of proliferating cell nuclear antigen (cyclin) in DNA replication in living cells. Mol. Cell. Biol. 9:57–66; 1989.
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Supported in part by grant No. HL 29351 from the National Institutes of Health, by a grant from the American Heart Association and with the support of the Southeastern Pennsylvania and Pennsylvania Affiliates of the American Heart Association.
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Marino, T.A., Walter, R.A., Cobb, E. et al. Effects of norepinephrine on neonatal rat cardiocyte growth and differentiation. In Vitro Cell Dev Biol 26, 229–236 (1990). https://doi.org/10.1007/BF02624452
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DOI: https://doi.org/10.1007/BF02624452