We show that remarkable differences exist among 3 mitochondrial subpopulations with regard to oxidation-phosphorylation coupling. In the heavy fraction (sedimenting at 3,000 g) ATP synthesis is optimized while in the medium and light fractions (sedimenting at 10,000 and 27,000 g, respectively) heat production tends to be optimized as the mitochondrial size decreases. As the oxygen consumption of the L-fraction drastically increases (+150%) after fifteen days of cold exposure, such differences have remarkable implications for the mechanism of cellular thermogenesis and its regulation.