The peripheral administration of streptozotocin (STZ) causes marked hyperglycaemia, plasma hyperosmolality, hypoinsulinaemia, hyperphagia and polydipsia. In STZ-induced diabetic animals, insulin treatment reverses these symptoms. Several studies have demonstrated that insulin deficiency induced diabetes causes alteration of hypothalamic transmitters and the gene expressions of neuropeptides [1±7]. These studies suggest that the alteration of monoamine and neuropeptides in the hypothalamus may play a role in the multiple behavioural and hormonal abnormalities in STZ-induced diabetic rats.Recent studies have demonstrated that nitric oxide (NO) may be associated with various functions of the central nervous system, including feeding [8±12] and drinking behaviour [13,14]. It has been demonstrated that both systemic administration of a competitive antagonist of NO synthase (NOS) and Diabetologia (1998) Summary Plasma arginine vasopressin (AVP) is known to be elevated in patients with uncontrolled insulin-dependent diabetes mellitus who have plasma hyperosmolality with hyperglycaemia. Although osmotic stimuli cause an increase in nitric oxide synthase (NOS) activity as well as synthesis of AVP and oxytocin in the paraventricular (PVN) and supraoptic nuclei (SON), it is not known whether NOS activity in the hypothalamus changes in the diabetic patients who have plasma hyperosmolality with hyperglycaemia caused by insulin deficiency. Expression of the neuronal (n) NOS gene in the PVN and SON in streptozotocin (STZ)-induced diabetic rats was investigated by using in situ hybridization histochemistry and NADPH-diaphorase histochemical staining. Four weeks after intraperitoneal (i. p.) administration of STZ, male Wistar rats developed hyperglycaemia and plasma hyperosmolality. The expression of nNOS gene and NADPH-diaphorase staining in the PVN and SON remarkably increased in STZ-induced diabetic rats compared to control rats. Three weeks after administration of STZ, the diabetic rats were subcutaneously treated with insulin for 1 week, which resulted in significant suppression of the induction of nNOS, AVP and oxytocin gene expression in the PVN and SON. Furthermore, the induction of nNOS gene expression in the PVN and SON was suppressed in STZ-induced diabetic rats treated with phlorizin and diet to normalize hyperglycaemia without insulin treatment. These results suggest that upregulation of nNOS gene expression as well as AVP and oxytocin gene expression in the PVN and SON in STZ-induced diabetic rats may be associated with hyperglycaemia and plamsa hyperosmolality. [Diabetologia (1998) 41: 640±648]