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R E Welikson

Publications and source records attributed to R E Welikson.

4 recordsLinked to original sources

Cardiac myosin heavy chains lacking the light chain binding domain cause hypertrophic cardiomyopathy in mice.

Myosin is a chemomechanical motor that converts chemical energy into the mechanical work of muscle contraction. More than 40 missense mutations in the cardiac myosin heavy chain (MHC) gene and several mutations in the two myosin light chains cause a dominantly inherited heart disease called familial hypertrophic cardiomyopathy. Very little is known about the biochemical defects in these alleles and how the mutations lead to disease. Because removal of the light chain binding domain in the lever arm of MHC should alter myosin's force transmission but not its catalytic function, we tested the hypothesis that such a mutant MHC would act as a dominant mutation in cardiac muscle. Hearts from transgenic mice expressing this mutant myosin are asymmetrically hypertrophied, with increases in mass primarily restricted to the cardiac anterior wall. Histological examination demonstrates marked cellular hypertrophy, myocyte disorganization, small vessel coronary disease, and severe valvular pathology that included thickening and plaque formation. Skinned myocytes and multicellular preparations from transgenic hearts exhibited decreased Ca2+ sensitivity of tension and decreased relaxation rates after flash photolysis of diazo 2. These experiments demonstrate that alterations in myosin force transmission are sufficient to trigger the development of hypertrophic cardiomyopathy.

Animals↗

Direct gene transfer into the mouse heart.

Direct injection of plasmid DNA into the myocardium of several species has been shown to be useful for studying cardiac gene expression. However, despite a better understanding of mouse genetics and the availability of several disease models in mice, gene injection with plasmid DNA into the mouse heart has not been reported. In this study, we demonstrate a simple and reproducible method for gene transfer into the mouse heart via direct injection of plasmid DNA. A firefly luciferase gene, driven by the RSV promoter, was used to quantitatively determine the spatial and temporal characteristics of gene transfer. Luciferase gene expression was stable for 8 weeks and showed a dose-dependent response over a range of 0.3-3 micrograms of input DNA. Inter-animal variability was low and gene expression was restricted to the left ventricle, near the site of injection. This method was also demonstrated to be suitable for detecting the expression of structural genes under the control of cellular promoters. Immunohistochemistry was used to detect the expression of an epitope-tagged myosin heavy chain driven by a rat alpha-myosin heavy chain promoter. Thus, naked DNA injection into the mouse heart results in a highly reproducible expression of constructs with either viral or cellular promoters. It is a relatively inexpensive and efficient means of studying cardiac gene regulation in vivo and a useful tool for screening the potential transgenes before generating transgenic mice.

Animals↗

The vertebrate myosin heavy chain: genetics and assembly properties.

The vertebrate sarcomere is a complex structure composed of numerous proteins arranged in an exquisitely precise manner. Sarcomeric proteins are organized into interdigitating thick or thin filaments and the sliding of these filaments relative to one another constitutes muscle contraction at the sarcomere level. Consequently, an understanding of sarcomeric structure and function requires a thorough knowledge of the individual components of the thick and thin filaments, as well as their associations. Thick filaments are comprised of myosin, which provides the force required to drive muscle contraction and also plays a major structural role in thick filament formation. In addition, a family of thick filament-associated proteins plays a role in organization of the thick filament. We have used both molecular genetic and cell biological approaches to define the diversity of the myosin heavy chain gene family and to analyze the assembly of myosin and it's associated proteins into thick filaments.

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