The increase in both glucose 6-phosphatase and hexokinase activities in brown adipose tissues of cold-exposed mice probably relates to thermogenesis by the substrate cycle between glucose 6-phosphate and glucose (Watanabe et al.: Anatomical Record 219:39-44, 1987). To clarify the factors causing the simultaneous increase, we examined biochemically the effects of uni- or bilateral surgical denervation of brown adipose tissues, of adrenalectomy, or of streptozotocin injection on the increase in the two activities in the tissues of cold-exposed mice. Further, the effects of denervation on the increase were also examined histochemically. The simultaneous increase in the two activities was inhibited in the denervated tissues of cold-exposed animals in biochemical and histochemical experiments. However, the increase in the activities was not inhibited in the tissues of animals exposed to cold after adrenalectomy or streptozotocin injection. The results suggest strongly that the activation of the substrate cycle in brown adipose tissues of cold-exposed mice is caused by a transmitter released from sympathetic nerve endings, probably norepinephrine.