title: Hypertrophic stimulation increases beta-actin dynamics in adult feline cardiomyocytes.
abstract: The myocardium responds to hemodynamic stress through cellular growth and organ hypertrophy. The impact of cytoskeletal elements on this process, however, is not fully understood. While alpha-actin in cardiomyocytes governs muscle contraction in combination with the myosin motor, the exact role of beta-actin has not been established. We hypothesized that in adult cardiomyocytes, as in non-myocytes, beta-actin can facilitate cytoskeletal rearrangement within cytoskeletal structures such as Z-discs. Using a feline right ventricular pressure overload (RVPO) model, we measured the level and distribution of beta-actin in normal and pressure overloaded myocardium. Resulting data demonstrated enriched levels of beta-actin and enhanced translocation to the Triton-insoluble cytoskeletal and membrane skeletal complexes. In addition, RVPO in vivo and in vitro hypertrophic stimulation with endothelin (ET) or insulin in isolated adult cardiomyocytes enhanced the content of polymerized fraction (F-actin) of beta-actin. To determine the localization and dynamics of beta-actin, we adenovirally expressed GFP-tagged beta-actin in isolated adult cardiomyocytes. The ectopically expressed beta-actin-GFP localized to the Z-discs, costameres, and cell termini. Fluorescence recovery after photobleaching (FRAP) measurements of beta-actin dynamics revealed that beta-actin at the Z-discs is constantly being exchanged with beta-actin from cytoplasmic pools and that this exchange is faster upon hypertrophic stimulation with ET or insulin. In addition, in electrically stimulated isolated adult cardiomyocytes, while beta-actin overexpression improved cardiomyocyte contractility, immunoneutralization of beta-actin resulted in a reduced contractility suggesting that beta-actin could be important for the contractile function of adult cardiomyocytes. These studies demonstrate the presence and dynamics of beta-actin in the adult cardiomyocyte and reinforce its usefulness in measuring cardiac cytoskeletal rearrangement during hypertrophic stimulation.
authors.affiliation: Cardiology Division, Department of Medicine, Gazes Cardiac Research Institute, Medical University of South Carolina, Charleston, South Carolina, United States of America. balasubr@musc.edu
authors.forename: Sundaravadivel
authors.initials: S
authors.lastname: Balasubramanian
authors.forename: Santhosh K
authors.initials: SK
authors.lastname: Mani
authors.forename: Harinath
authors.initials: H
authors.lastname: Kasiganesan
authors.forename: Catalin C
authors.initials: CC
authors.lastname: Baicu
authors.forename: Dhandapani
authors.initials: D
authors.lastname: Kuppuswamy
mesh_terms: D000199:Actins / Q000378:metabolism*; D000818:Animals; D015153:Blotting, Western; D002415:Cats; D002478:Cells, Cultured; D036681:Fluorescence Recovery After Photobleaching; D007150:Immunohistochemistry; D032383:Myocytes, Cardiac / Q000378:metabolism* / Q000473:pathology*; D020033:Protein Isoforms / Q000378:metabolism*; D018497:Ventricular Dysfunction, Right / Q000378:metabolism; D017725:Ventricular Pressure / Q000502:physiology
chemical_list: D000199:Actins; D020033:Protein Isoforms