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Contractile protein gene expression in serum-free cultured adult rat cardiac myocytes

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Abstract

The effects of two adhesion substrates (serum and laminin) and time in culture on the expression of genes encoding myosin heavy chain (MHC) isoforms and α-skeletal actin were analysed in myocytes isolated from adult rat heart and maintained in serum-free culture. Relative messenger ribonucleic acid (mRNA) abundances were quantitated by dot-blot analysis. Gene expression was not influenced by the substrate used. Time in culture induced a decrease in total mRNA abundance and an up-regulation of β-MHC and α-skeletal actin genes. It is proposed that atrophy of adult myocytes is associated with a pattern of gene expression similar to the fetal program.

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References

  1. Alonso S, Garner I, Vandekerckhove J, Buckingham M (1990) Genetic analysis of the interaction between cardiac and skeletal actin gene expression in striated muscle of the mouse, J Mol Biol 211:727–738

    Google Scholar 

  2. Bergmeyer HU, Bernt E (1974) UV-assay for lactate dehydrogenase with pyruvate and NADH. In: Bergmeyer HU (ed) Methods of enzymatic analysis, vol 2. Verlag Chemie, Weinheim, pp 574–579

    Google Scholar 

  3. Boheler KRB, Carrier L, de la Bastie D, Allen PD, Komajda M, Mercadier JJ, Schwartz K (1991) Skeletal actin mRNA increases in the human heart during ontogenic development and is the major isoform of control and failing adult hearts. J Clin Invest 88:323–330

    Google Scholar 

  4. Borg TK, Rubin K, Lundgren E, Borg K, Öbrink B (1984) Recognition of extracellular matrix components by neonatal and adult cardiac myocytes. Dev Biol 104:86–96

    Google Scholar 

  5. Bugaisky LB, Zak R (1989) Differentiation of adult cardiac myocytes in cell culture. Circ Res 64:493–500

    Google Scholar 

  6. Carrier L, Boheler KR, Chassagne C, de la Bastie D, Wisnewsky C, Lakatta EG, Schwartz K (1992) Expression of the sarcomeric actin isogenes in the rat heart with development and senescence. Circ Res 70:999–1005

    Google Scholar 

  7. Chomczynski P, Sacchi N (1987) Single-step method of RNA isolation by acid guanidium thiocyanate-phenol-chloroform extraction. Biochem 162:156–159

    Google Scholar 

  8. Delcayre C, Samuel JL, Marotte F, Best-Belpomme M, Rappaport L (1988) Synthesis of stress proteins is transiently increased in rat cardiac myocytes 2–4 days after imposition of of hemodynamic overload. J Clin Invest 2:460–468

    Google Scholar 

  9. Dubus I, Samuel JL, Marotte F, Delcayre C, Rappaport L (1990) β-Adrenergic agonists stimulate the synthesis of noncontractile but not contractile proteins in cultured myocytes isolated from adult rat heart. Circ Res 66:867–874

    Google Scholar 

  10. Dubus I, Nallet O, Mercadier A, Lucas O, Rappaport L, Samuel JL (1992) Heterogeneous distribution of α- and β-MHC mRNAs within adult rat cardiac myocyte population: a quantitative in situ hybridization study (abstract). J Mol Cell Cardiol 24 [Suppl 5]: S 11

    Google Scholar 

  11. Eppenberger-Eberhardt M, Flamme I, Kurer V, Eppenberger HM (1990) Reexpression of α-smooth muscle actin isoform in cultured adult rat cardiomyocytes. Dev Biol 139:269–278

    Google Scholar 

  12. Harley CB (1987) Hybridization of oligo(dT) to RNA on nitrocellulose. Genet Anal Techn 4:17–22

    Google Scholar 

  13. Klein L, Omajaa K, Samarel AM, Welikson R, Hong C (1992) Hemodynamic regulation of myosin heavy chain gene expression; studies in the transplanted rat heart. J Clin Invest 89:68–73

    Google Scholar 

  14. Kühl U, Öcalan M, Timpl R, von der Mark K (1986) Role of laminin and fibronectin in selecting myogenic versus fibrogenic cells from skeletal muscle cells in vitro. Dev Biol 117:628–635

    Google Scholar 

  15. Lompré AM, Nadal-Ginard B, Mahdavi V (1984) Expression of the cardiac ventricular α- and β-myosin heavy chain genes is developmentally and hormonally regulated. J Biol Chem 259:6437–6446

    Google Scholar 

  16. Lundgren E, Terracio L, Mardh S, Borg TK (1985) Extracellular matrix components influence the survival of adult cardiac myocytes in vitro. Exp Cell Res 158:371–381

    Google Scholar 

  17. Mayer Y, Czosnek H, Zeelon PE, Yaffe D, Nudel U (1984) Expression of the genes coding for the skeletal muscle and cardiac actins in the heart. Nucleic Acids Res 12:1087–1100

    Google Scholar 

  18. McDermott PJ, Morgan HE (1989) Contraction modulates the capacity for protein synthesis during growth of neonatal heart cells in culture. Circ Res 64:542–553

    Google Scholar 

  19. Mercadier JJ, Dubus I (1991) Assay of atrial natriuretic factor messenger ribonucleic acid. Methods Neurosci 5:22–34

    Google Scholar 

  20. Mercadier JJ, Samuel JL, Michel JB, Zongazo MA, de la Bastie D, Lompré AM, Wisnewsky C, Rappaport L, Levy B, Schwartz K (1989) Atrial natriuretic factor gene expression in rat ventricle during experimental hypertension. Am J Physiol 257:H 979-H 987

    Google Scholar 

  21. Piper HM, Voltz A, Schwartz P (1990) Adult ventricular rat heart cells. In: Piper HM (ed) Cell culture techniques in heart and vessel research. Springer, Berlin Heidelberg New York, pp 36–60

    Google Scholar 

  22. Samarel AM, Engelmann GL (1991) Contractile activity modulates myosin heavy chain-β expression in neonatal rat heart cells. Am J Physiol 261:H 1067-H 1077

    Google Scholar 

  23. Schiaffino S, Samuel JL, Sassoon D, Lompré AM, Garner L, Marotte F, Buckingham M, Rappaport L, Schwartz K (1989) Nonsynchronous accumulation of α-skeletal actin and β-myosin heavy chain mRNAs during early stages of pressure overloaded-induced cardiac hypertrophy demonstrated by in situ hybridization. Circ Res 64:937–948

    Google Scholar 

  24. Schneider MD, Parker TG (1990) Cardiac myocytes as targets for the action of peptide growth factors. Circulation 81:1443–1456

    Google Scholar 

  25. Simpson P (1985) Stimulation of hypertrophy of cultured neonatal rat heart cells through an α1-adrenergic receptor and induction of beating through an α1- and β1-adrenergic receptor interaction. Circ Res 56:884–894

    Google Scholar 

  26. Smith WR, Claycomb WC (1989) Expression of α- and β-myosin heavy chain genes in the rat ventricular cardiac muscle cells. In: Kedes LH, Stockdale FE (eds) Cellular and molecular biology of muscle development. Liss, New York, pp 359–368

    Google Scholar 

  27. Vusse GJ van der, Coumans WA, Veen FH van der, Drake A, Flameng W, Syy R (1984) ATP, creatine phosphate and glycogen content in human myocardial biopsies: markers for the efficacy of cardioprotection during aorta-coronary bypass surgery. Vase Surg 18:127–134

    Google Scholar 

  28. Winegrad S, Wisnewsky C, Schwartz K (1990) Effect of thyroid hormone on the accumulation of mRNA for skeletal and cardiac α-actin in hearts from normal and hypophysectomized rats. Proc Natl Acad Sci USA 87:2456–2460

    Google Scholar 

  29. Zakut R, Shani M, Givol D, Neuman S, Yaffe D, Nudel U (1982) Nucleotide sequence of the rat skeletal muscle actin gene. Nature 298:857–859

    Google Scholar 

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Authors and Affiliations

  1. Unité 127 INSERM, Hôpital Lariboisière, 41 Bd. de la Chapelle, F-75010, Paris, France

    I. Dubus, L. Rappaport, A. Barrieux, K. Schwartz & J. L. Samuel

  2. Unité INSERM 275, Ecole Polytechnique, F-91120, Palaiseau, France

    A. M. Lompré

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  1. I. Dubus
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  2. L. Rappaport
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  3. A. Barrieux
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  4. A. M. Lompré
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  5. K. Schwartz
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  6. J. L. Samuel
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Dubus, I., Rappaport, L., Barrieux, A. et al. Contractile protein gene expression in serum-free cultured adult rat cardiac myocytes. Pflügers Arch. 423, 455–461 (1993). https://doi.org/10.1007/BF00374941

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  • DOI: https://doi.org/10.1007/BF00374941

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