To investigate whether differences in metabolic rate are related to differences in muscle mitochondrial morphology and/or to differences in in vitro muscle respiration, we studied 17 healthy Caucasians, covering a wide range of body weight and composition [9 males, 8 females; body wt 96 +/- 37 (SD) kg; body fat = 28 +/- 10%]. Central and peripheral mitochondrial volume density (Vmit c and Vmit p, respectively) and the ratio of mitochondrial outer surface to volume of mitochondria (SVmit c in center and SVmit p at periphery) were determined by stereological analyses of transmission electron micrographs from samples of the vastus lateralis. There was no relationship between mitochondrial morphology or muscle respiration and 24-h energy expenditure, basal metabolic rate, or sleeping energy expenditure adjusted for differences in fat-free mass, fat mass, age, and sex. Although total body fat was not associated with muscle cell morphology, central distribution of body fat [waist-to-thigh circumference ratio (W/T)] correlated negatively with Vmit c (r = -0.58, P = 0.01), SVmit c (r = -0.59, P = 0.01), and SVmit p (r = -0.48, P = 0.05). W/T was also negatively related to muscle respiration (r = -0.59, P = 0.01). Despite the lack of relationship between metabolic rate and muscle mitochondrial morphology, central distribution of body fat is associated with lower mitochondrial density and larger mitochondria in skeletal muscle and is associated with a decreased oxidative capacity of muscle.