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This study examined the effects of raising muscle glycogen by carbohydrate feeding and of keeping muscle glycogen low by carbohydrate restriction following exhausting exercise on the ability of perfused skeletal muscle to take up glucose and to synthesize glycogen. Muscle glycogen concentration was more than twice as high in the rats fed carbohydrate as in those not given carbohydrate. Muscle glycogen synthesis during a 30-min perfusion with glucose and insulin was significantly greater in the animals with low muscle glycogen. Furthermore the muscles with low glycogen content converted a greater proportion of the glucose taken up to glycogen and less to lactate than did the muscles with high glycogen content. In rats subjected to exhausting exercise on the preceding day, the rate of glucose uptake by perfused skeletal muscle was significantly higher (60-80%) at the same insulin concentration in animals in which muscle glycogen was kept low than in those in which glycogen was raised by carbohydrate feeding.