Cardiac transgenesis with the tetracycline transactivator changes myocardial function and gene expression

Diana T McCloskey, Lynne Turnbull, Philip M Swigart, Alexander C Zambon, Sally Turcato, Shuji Joho, William Grossman, Bruce R Conklin, Paul C Simpson, Anthony J Baker

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Abstract

The cardiac-specific tetracycline-regulated gene expression system (tet-system) is a powerful tool using double-transgenic mice. The cardiac alpha-myosin heavy chain promoter (alphaMHC) drives lifetime expression of a tetracycline-inhibited transcription activator (tTA). Crossing alphaMHC-tTA mice with mice containing a tTA-responsive promoter linked to a target gene yields double-transgenic mice having tetracycline-repressed expression of the target gene in the heart. Using the tet-system, some studies use nontransgenic mice for the control group, whereas others use single-transgenic alphaMHC-tTA mice. However, previous studies found that high-level expression of a modified activator protein caused cardiomyopathy. Therefore, we tested whether cardiac expression of tTA was associated with altered function of alphaMHC-tTA mice compared with wild-type (WT) littermates. We monitored in vivo and in vitro function and gene expression profiles for myocardium from WT and alphaMHC-tTA mice. Compared with WT littermates, alphaMHC-tTA mice had a greater heart-to-body weight ratio (approximately 10 ), ventricular dilation, and decreased ejection fraction, suggesting mild cardiomyopathy. In vitro, submaximal contractions were greater compared with WT and were associated with greater myofilament Ca2+ sensitivity. Gene expression profiling revealed that the expression of 153 genes was significantly changed by >20 when comparing alphaMHC-tTA with WT myocardium. These findings demonstrate that introduction of the alphaMHC-tTA construct causes significant effects on myocardial gene expression and major functional abnormalities in vivo and in vitro. For studies using the tet-system, these results suggest caution in the use of controls, since alphaMHC-tTA myocardium differs appreciably from WT. Furthermore, the results raise the possibility that the phenotype conferred by a target gene may be influenced by the modified genetic background of alphaMHC-tTA myocardium.
Original languageEnglish
Pages (from-to)118 - 126
Number of pages9
JournalPhysiological Genomics
Volume22
Issue number1
Publication statusPublished - 2005

Cite this

McCloskey, D. T., Turnbull, L., Swigart, P. M., Zambon, A. C., Turcato, S., Joho, S., Grossman, W., Conklin, B. R., Simpson, P. C., & Baker, A. J. (2005). Cardiac transgenesis with the tetracycline transactivator changes myocardial function and gene expression. Physiological Genomics, 22(1), 118 - 126.