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J. Biol. Chem., Vol. 278, Issue 18, 15927-15934, May 2, 2003

This Article Full Text Full Text (PDF) Data Supplement All Versions of this Article: 278/18/15927    most recent M213102200v1 Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Rybkin, I. I. Articles by Olson, E. N. Search for Related Content PubMed PubMed Citation Articles by Rybkin, I. I. Articles by Olson, E. N. Social Bookmarking                      What's this?

Conditional Expression of SV40 T-antigen in Mouse Cardiomyocytes Facilitates an Inducible Switch from Proliferation to Differentiation^*,

Igor I. Rybkin, David W. Markham, Zhen Yan, Rhonda Bassel-Duby, R. Sanders Williams, and Eric N. Olson^§

From the Departments of Molecular Biology and Internal Medicine, University of Texas Southwestern Medical Center, Dallas, Texas 75390 and  Duke University Medical School, Durham, North Carolina 27710

Studies of cardiac muscle gene expression and signaling have been hampered by the lack of immortalized cardiomyocyte cell lines capable of proliferation and irreversible withdrawal from the cell cycle. With the goal of creating such cell lines, we generated transgenic mice using cardiac-specific cis-regulatory elements from the mouse Nkx2.5 gene to drive the expression of a simian virus 40 large T-antigen (TAg) gene flanked by sites for recombination by Cre recombinase. These transgenic mice developed tumors within the ventricular myocardium. Cells isolated from these tumors expressed cardiac markers and proliferated rapidly during serial passage in culture, without apparent senescence. However, they were unable to exit the cell cycle and failed to exhibit morphological features of terminal differentiation. Introduction of Cre recombinase to these cardiac cell lines by adenoviral delivery resulted in the elimination of TAg expression, accompanied by rapid cessation of cell division, and increase in cell size without an apparent induction of cellular differentiation. Incubation of cells lacking TAg in serum-deficient media with various pharmacological agents (norepinephrine, phenylephrine, or bone morphogenetic protein-2/4) or constitutively active calcium/calmodulin-dependent protein kinase I and/or calcineurin led to the formation of sarcomeres and up-regulation of cardiac genes involved in excitation-contraction coupling. The combination of TAg expression under the control of an early cardiac promoter and Cre-mediated recombination allowed us to derive an immortal cell line from the ventricular myocardium that could be controllably withdrawn from the cell cycle. The conditional expression of TAg in this manner permits propagation and regulated growth termination of cell types that are otherwise unable to be maintained in cell culture and may have applications for cardiac repair technologies.

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