ACTIVATION OF GLUT4 IN HYPOXIC CARDIAC MYOCYTES

Yale University, New Haven CT

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Abstract

DESCRIPTION The applicant aims to understand the molecular physiologic responses to myocardial ischemia to develop novel metabolic approaches to treat patients with coronary heart disease. The first aim will be to evaluate 5'- AMP activated protein kinase (AMPK) in hypoxic cardiomyocytes and to study the effects of AMPK in regulating glucose transport and protein kinases (AMPK) in hypoxic cardiomyocytes and to study the effects of AMPK in regulating glucose transport and utilization. These studies will utilize biochemical measurements of sarcolemmal GLUT4, immunofluorescence, glucose transport kinetics and stable isotope and radiotracer measurements of glycolytic flux. The second aim will involve studying the kinetics of GLUT4 to examine the contributions of exocytosis and endocytosis during hypoxia to test the hypothesis that inhibition of endocytosis will play a significant role in increasing sarcolemmal GLUT4 content during hypoxia. Aim 3 will examine whether dynamin is involved in GLUT4 endocytosis in cardiac myocytes and whether ischemia and AMPK modulate sarcolemma GLUT by influencing dynamin. These experiments will involve the expression of a dominant negative dynamin mutant in cultured cardiomyocytes after adenovirus transfection. These studies are anticipated to provide the applicant with expertise in mechanisms involved in the cellular response to hypoxia and the tools to initiate an independent investigative career.