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. 1997 Jun 1;99(11):2644–2654. doi: 10.1172/JCI119453

Cyclin D1 overexpression promotes cardiomyocyte DNA synthesis and multinucleation in transgenic mice.

M H Soonpaa 1, G Y Koh 1, L Pajak 1, S Jing 1, H Wang 1, M T Franklin 1, K K Kim 1, L J Field 1
PMCID: PMC508110  PMID: 9169494

Abstract

D-type cyclin/cyclin-dependent kinase (CDK) complexes regulate transit through the restriction point of the cell cycle, and thus are required for the initiation of DNA synthesis. Transgenic mice which overexpress cyclin D1 in the heart were produced to determine if D-type cyclin deregulation would alter myocardial development. Cyclin D1 overexpression resulted in a concomitant increase in CDK4 levels in the adult myocardium, as well as modest increases in proliferating cell nuclear antigen and CDK2 levels. Flow cytometric and morphologic analyses of dispersed cell preparations indicated that the adult transgenic cardiomyocytes had abnormal patterns of multinucleation. Histochemical analyses confirmed a marked increase in number of cardiomyocyte nuclei in sections prepared from the transgenic mice as compared with those from control animals. Tritiated thymidine incorporation analyses revealed sustained cardiomyocyte DNA synthesis in adult transgenic hearts.

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Selected References

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  1. Anversa P., Kajstura J., Reiss K., Quaini F., Baldini A., Olivetti G., Sonnenblick E. H. Ischemic cardiomyopathy: myocyte cell loss, myocyte cellular hypertrophy, and myocyte cellular hyperplasia. Ann N Y Acad Sci. 1995 Mar 27;752:47–64. doi: 10.1111/j.1749-6632.1995.tb17405.x. [DOI] [PubMed] [Google Scholar]
  2. Beijersbergen R. L., Carlée L., Kerkhoven R. M., Bernards R. Regulation of the retinoblastoma protein-related p107 by G1 cyclin complexes. Genes Dev. 1995 Jun 1;9(11):1340–1353. doi: 10.1101/gad.9.11.1340. [DOI] [PubMed] [Google Scholar]
  3. Bodrug S. E., Warner B. J., Bath M. L., Lindeman G. J., Harris A. W., Adams J. M. Cyclin D1 transgene impedes lymphocyte maturation and collaborates in lymphomagenesis with the myc gene. EMBO J. 1994 May 1;13(9):2124–2130. doi: 10.1002/j.1460-2075.1994.tb06488.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Daud A. I., Lanson N. A., Jr, Claycomb W. C., Field L. J. Identification of SV40 large T-antigen-associated proteins in cardiomyocytes from transgenic mice. Am J Physiol. 1993 May;264(5 Pt 2):H1693–H1700. doi: 10.1152/ajpheart.1993.264.5.H1693. [DOI] [PubMed] [Google Scholar]
  5. Deng C., Zhang P., Harper J. W., Elledge S. J., Leder P. Mice lacking p21CIP1/WAF1 undergo normal development, but are defective in G1 checkpoint control. Cell. 1995 Aug 25;82(4):675–684. doi: 10.1016/0092-8674(95)90039-x. [DOI] [PubMed] [Google Scholar]
  6. Dowdy S. F., Hinds P. W., Louie K., Reed S. I., Arnold A., Weinberg R. A. Physical interaction of the retinoblastoma protein with human D cyclins. Cell. 1993 May 7;73(3):499–511. doi: 10.1016/0092-8674(93)90137-f. [DOI] [PubMed] [Google Scholar]
  7. Engelmann G. L., Birchenall-Roberts M. C., Ruscetti F. W., Samarel A. M. Formation of fetal rat cardiac cell clones by retroviral transformation: retention of select myocyte characteristics. J Mol Cell Cardiol. 1993 Feb;25(2):197–213. doi: 10.1006/jmcc.1993.1022. [DOI] [PubMed] [Google Scholar]
  8. Ewen M. E., Sluss H. K., Sherr C. J., Matsushime H., Kato J., Livingston D. M. Functional interactions of the retinoblastoma protein with mammalian D-type cyclins. Cell. 1993 May 7;73(3):487–497. doi: 10.1016/0092-8674(93)90136-e. [DOI] [PubMed] [Google Scholar]
  9. Field L. J. Transgenic mice in cardiovascular research. Annu Rev Physiol. 1993;55:97–114. doi: 10.1146/annurev.ph.55.030193.000525. [DOI] [PubMed] [Google Scholar]
  10. Graña X., Reddy E. P. Cell cycle control in mammalian cells: role of cyclins, cyclin dependent kinases (CDKs), growth suppressor genes and cyclin-dependent kinase inhibitors (CKIs). Oncogene. 1995 Jul 20;11(2):211–219. [PubMed] [Google Scholar]
  11. Gulick J., Subramaniam A., Neumann J., Robbins J. Isolation and characterization of the mouse cardiac myosin heavy chain genes. J Biol Chem. 1991 May 15;266(14):9180–9185. [PubMed] [Google Scholar]
  12. Hunter T., Pines J. Cyclins and cancer. II: Cyclin D and CDK inhibitors come of age. Cell. 1994 Nov 18;79(4):573–582. doi: 10.1016/0092-8674(94)90543-6. [DOI] [PubMed] [Google Scholar]
  13. Jackson T., Allard M. F., Sreenan C. M., Doss L. K., Bishop S. P., Swain J. L. The c-myc proto-oncogene regulates cardiac development in transgenic mice. Mol Cell Biol. 1990 Jul;10(7):3709–3716. doi: 10.1128/mcb.10.7.3709. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Jackson T., Allard M. F., Sreenan C. M., Doss L. K., Bishop S. P., Swain J. L. Transgenic animals as a tool for studying the effect of the c-myc proto-oncogene on cardiac development. 1991 May 29-Jun 12Mol Cell Biochem. 104(1-2):15–19. doi: 10.1007/BF00229798. [DOI] [PubMed] [Google Scholar]
  15. Jiang W., Kahn S. M., Zhou P., Zhang Y. J., Cacace A. M., Infante A. S., Doi S., Santella R. M., Weinstein I. B. Overexpression of cyclin D1 in rat fibroblasts causes abnormalities in growth control, cell cycle progression and gene expression. Oncogene. 1993 Dec;8(12):3447–3457. [PubMed] [Google Scholar]
  16. Jin J. P., Lin J. J. Isolation and characterization of cDNA clones encoding embryonic and adult isoforms of rat cardiac troponin T. J Biol Chem. 1989 Aug 25;264(24):14471–14477. [PubMed] [Google Scholar]
  17. Kim K. K., Soonpaa M. H., Daud A. I., Koh G. Y., Kim J. S., Field L. J. Tumor suppressor gene expression during normal and pathologic myocardial growth. J Biol Chem. 1994 Sep 9;269(36):22607–22613. [PubMed] [Google Scholar]
  18. Kirshenbaum L. A., Schneider M. D. Adenovirus E1A represses cardiac gene transcription and reactivates DNA synthesis in ventricular myocytes, via alternative pocket protein- and p300-binding domains. J Biol Chem. 1995 Apr 7;270(14):7791–7794. doi: 10.1074/jbc.270.14.7791. [DOI] [PubMed] [Google Scholar]
  19. Kiyokawa H., Busquets X., Powell C. T., Ngo L., Rifkind R. A., Marks P. A. Cloning of a D-type cyclin from murine erythroleukemia cells. Proc Natl Acad Sci U S A. 1992 Mar 15;89(6):2444–2447. doi: 10.1073/pnas.89.6.2444. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  21. Lanson N. A., Jr, Glembotski C. C., Steinhelper M. E., Field L. J., Claycomb W. C. Gene expression and atrial natriuretic factor processing and secretion in cultured AT-1 cardiac myocytes. Circulation. 1992 May;85(5):1835–1841. doi: 10.1161/01.cir.85.5.1835. [DOI] [PubMed] [Google Scholar]
  22. Lovec H., Grzeschiczek A., Kowalski M. B., Möröy T. Cyclin D1/bcl-1 cooperates with myc genes in the generation of B-cell lymphoma in transgenic mice. EMBO J. 1994 Aug 1;13(15):3487–3495. doi: 10.1002/j.1460-2075.1994.tb06655.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Matsushime H., Roussel M. F., Ashmun R. A., Sherr C. J. Colony-stimulating factor 1 regulates novel cyclins during the G1 phase of the cell cycle. Cell. 1991 May 17;65(4):701–713. doi: 10.1016/0092-8674(91)90101-4. [DOI] [PubMed] [Google Scholar]
  24. Motokura T., Keyomarsi K., Kronenberg H. M., Arnold A. Cloning and characterization of human cyclin D3, a cDNA closely related in sequence to the PRAD1/cyclin D1 proto-oncogene. J Biol Chem. 1992 Oct 5;267(28):20412–20415. [PubMed] [Google Scholar]
  25. Quelle D. E., Ashmun R. A., Shurtleff S. A., Kato J. Y., Bar-Sagi D., Roussel M. F., Sherr C. J. Overexpression of mouse D-type cyclins accelerates G1 phase in rodent fibroblasts. Genes Dev. 1993 Aug;7(8):1559–1571. doi: 10.1101/gad.7.8.1559. [DOI] [PubMed] [Google Scholar]
  26. Rao S. S., Chu C., Kohtz D. S. Ectopic expression of cyclin D1 prevents activation of gene transcription by myogenic basic helix-loop-helix regulators. Mol Cell Biol. 1994 Aug;14(8):5259–5267. doi: 10.1128/mcb.14.8.5259. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Rao S. S., Kohtz D. S. Positive and negative regulation of D-type cyclin expression in skeletal myoblasts by basic fibroblast growth factor and transforming growth factor beta. A role for cyclin D1 in control of myoblast differentiation. J Biol Chem. 1995 Feb 24;270(8):4093–4100. doi: 10.1074/jbc.270.8.4093. [DOI] [PubMed] [Google Scholar]
  28. Raynal F., Michot B., Bachellerie J. P. Complete nucleotide sequence of mouse 18 S rRNA gene: comparison with other available homologs. FEBS Lett. 1984 Feb 27;167(2):263–268. doi: 10.1016/0014-5793(84)80139-8. [DOI] [PubMed] [Google Scholar]
  29. Reddy V. B., Thimmappaya B., Dhar R., Subramanian K. N., Zain B. S., Pan J., Ghosh P. K., Celma M. L., Weissman S. M. The genome of simian virus 40. Science. 1978 May 5;200(4341):494–502. doi: 10.1126/science.205947. [DOI] [PubMed] [Google Scholar]
  30. Reiss K., Cheng W., Ferber A., Kajstura J., Li P., Li B., Olivetti G., Homcy C. J., Baserga R., Anversa P. Overexpression of insulin-like growth factor-1 in the heart is coupled with myocyte proliferation in transgenic mice. Proc Natl Acad Sci U S A. 1996 Aug 6;93(16):8630–8635. doi: 10.1073/pnas.93.16.8630. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Resnitzky D., Gossen M., Bujard H., Reed S. I. Acceleration of the G1/S phase transition by expression of cyclins D1 and E with an inducible system. Mol Cell Biol. 1994 Mar;14(3):1669–1679. doi: 10.1128/mcb.14.3.1669. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Robbins J., Gulick J., Sanchez A., Howles P., Doetschman T. Mouse embryonic stem cells express the cardiac myosin heavy chain genes during development in vitro. J Biol Chem. 1990 Jul 15;265(20):11905–11909. [PubMed] [Google Scholar]
  33. Saule S., Mérigaud J. P., Al-Moustafa A. E., Ferré F., Rong P. M., Amouyel P., Quatannens B., Stéhelin D., Dieterlen-Lièvre F. Heart tumors specifically induced in young avian embryos by the v-myc oncogene. Proc Natl Acad Sci U S A. 1987 Nov;84(22):7982–7986. doi: 10.1073/pnas.84.22.7982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Serrano M., Lee H., Chin L., Cordon-Cardo C., Beach D., DePinho R. A. Role of the INK4a locus in tumor suppression and cell mortality. Cell. 1996 Apr 5;85(1):27–37. doi: 10.1016/s0092-8674(00)81079-x. [DOI] [PubMed] [Google Scholar]
  35. Skapek S. X., Rhee J., Spicer D. B., Lassar A. B. Inhibition of myogenic differentiation in proliferating myoblasts by cyclin D1-dependent kinase. Science. 1995 Feb 17;267(5200):1022–1024. doi: 10.1126/science.7863328. [DOI] [PubMed] [Google Scholar]
  36. Soonpaa M. H., Field L. J. Assessment of cardiomyocyte DNA synthesis during hypertrophy in adult mice. Am J Physiol. 1994 Apr;266(4 Pt 2):H1439–H1445. doi: 10.1152/ajpheart.1994.266.4.H1439. [DOI] [PubMed] [Google Scholar]
  37. Soonpaa M. H., Field L. J. Assessment of cardiomyocyte DNA synthesis in normal and injured adult mouse hearts. Am J Physiol. 1997 Jan;272(1 Pt 2):H220–H226. doi: 10.1152/ajpheart.1997.272.1.H220. [DOI] [PubMed] [Google Scholar]
  38. Soonpaa M. H., Kim K. K., Pajak L., Franklin M., Field L. J. Cardiomyocyte DNA synthesis and binucleation during murine development. Am J Physiol. 1996 Nov;271(5 Pt 2):H2183–H2189. doi: 10.1152/ajpheart.1996.271.5.H2183. [DOI] [PubMed] [Google Scholar]
  39. Soonpaa M. H., Koh G. Y., Klug M. G., Field L. J. Formation of nascent intercalated disks between grafted fetal cardiomyocytes and host myocardium. Science. 1994 Apr 1;264(5155):98–101. doi: 10.1126/science.8140423. [DOI] [PubMed] [Google Scholar]
  40. Steinhelper M. E., Cochrane K. L., Field L. J. Hypotension in transgenic mice expressing atrial natriuretic factor fusion genes. Hypertension. 1990 Sep;16(3):301–307. doi: 10.1161/01.hyp.16.3.301. [DOI] [PubMed] [Google Scholar]
  41. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Wang T. C., Cardiff R. D., Zukerberg L., Lees E., Arnold A., Schmidt E. V. Mammary hyperplasia and carcinoma in MMTV-cyclin D1 transgenic mice. Nature. 1994 Jun 23;369(6482):669–671. doi: 10.1038/369669a0. [DOI] [PubMed] [Google Scholar]
  43. Xiong Y., Zhang H., Beach D. D type cyclins associate with multiple protein kinases and the DNA replication and repair factor PCNA. Cell. 1992 Oct 30;71(3):505–514. doi: 10.1016/0092-8674(92)90518-h. [DOI] [PubMed] [Google Scholar]
  44. Zivin R. A., Condra J. H., Dixon R. A., Seidah N. G., Chrétien M., Nemer M., Chamberland M., Drouin J. Molecular cloning and characterization of DNA sequences encoding rat and human atrial natriuretic factors. Proc Natl Acad Sci U S A. 1984 Oct;81(20):6325–6329. doi: 10.1073/pnas.81.20.6325. [DOI] [PMC free article] [PubMed] [Google Scholar]

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