Changes inβ-actin mRNA Expression in Remodeling Canine Myocardium

doi:10.1006/jmcc.1996.0006

Journal of Molecular and Cellular Cardiology

Volume 28, Issue 1, 8 January 1996, Pages 53-63

https://doi.org/10.1006/jmcc.1996.0006 Get rights and content

Abstract

Beta-actin, a cytoskeletal protein important in the maintenance of cytoarchitecture, has long been thought to be expressed constitutively in myocardial tissue. As such,β-actin mRNA has been used as a control gene in a wide range of experiments. However, we have uncovered consistent changes inβ-actin mRNA expression in canine myocardium remodeling as a result of insult to the left ventricle. The experimental canine models used were either DC shock damage to the left ventricle or volume overload resulting from severe mitral regurgitation. The remodeling process in both canine models is characterized by an increase in left ventricular mass. PCR amplification using primers designed to selectively amplify the 3 ′ end and a portion of the 3 ′ untranslated region ofβ-actin mRNA resulted in the generation of a 297 base pair product predominant only in normal canine myocardium and a 472 base pair product that became increasingly prominent from 1 to 30 days after DC shock damage to the left ventricle and from 10 to 90 days after creation of mitral regurgitation. Northern analysis showed a three-fold increase inβ-actin mRNA after either DC shock or creation of mitral regurgitation. Western analysis revealed an early increase inβ-actin protein followed by an apparent decrease to below baseline levels. These observations suggest that changes inβ-actin mRNA expression accompany the structural alterations that occur in response to myocardial damage. Whether or not the changes inβ-actin mRNA expression play a role in mediating these structural alterations remains to be determined.

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Among these studies, β-actin and 18S rRNA have been commonly used as internal standards without validation for its appropriateness. Contrary evidences have suggested that β-actin is inappropriate in some cases (Carlyle et al., 1996; Glare et al., 2002; Zheng and Sun, 2011). Previous studies have also suggested that cautions should be paid when using 18S rRNA as a reference control (Tang et al., 2007).
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^☆: Please address all correspondence to: Jay N. Cohn, Cardiovascular Division, University of Minnesota Medical School, Box 508 UMHC, 420 Delaware Street SE, Minneapolis, MN 55455, U.S.A.

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Regular paperChanges inβ-actin mRNA Expression in Remodeling Canine Myocardium☆

Abstract

Regular paper
Changes inβ-actin mRNA Expression in Remodeling Canine Myocardium☆