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Obesity is associated with oxidative stress and mitochondrial and myocardial dysfunction, although interaction among which remains elusive. This study was designed to evaluate the impact of the free radical scavenger metallothionein on high-fat diet–induced myocardial, intracellular Ca²⁺, and mitochondrial dysfunction. FVB and metallothionein transgenic mice were fed a high- or low-fat diet for 5 months to induce obesity. Echocardiography revealed decreased fractional shortening, increased end-systolic diameter, and cardiac hypertrophy in high-fat–fed FVB mice. Cardiomyocytes from high-fat–fed FVB mice displayed enhanced reactive oxygen species (ROS) production, contractile and intracellular Ca²⁺ defects including depressed peak shortening and maximal velocity of shortening/relengthening, prolonged duration of relengthening, and reduced intracellular Ca²⁺ rise and clearance. Transmission …