We determined the contribution of central N-methyl-D-aspartate (NMDA) receptor activation to the neuro-endocrine counter-regulatory response to insulin-induced hypoglycemia. Glucose kinetics, gluconeogenic substrate balance and counter-regulatory hormonal responses were determined in two groups of conscious dogs fitted with chronic vascular catheters and intracerebroventricular (i.c.v.) cannula. Peripheral insulin infusion (5 mU/kg per min for 3 h) decreased plasma glucose levels 40% and increased the rate of glucose appearance (R(a)) 2-fold. This was associated with significant increases in net hepatic uptake of glycerol and lactate, without any change in the net hepatic uptake of alanine. i.c.v. pretreatment with MK-801, an NMDA receptor antagonist, blunted (50%) the rise in glucose R(a) as well as the increase in the net hepatic uptake of glycerol and lactate. Hypoglycemia increased plasma cortisol (3-fold to 14.3+/-1 mg/dl) and epinephrine levels (14-fold to 3811+/-172 pg/ml), and this stress response was attenuated (30% and 60%, respectively) by MK-801 pretreatment. In controls, MK-801 did not alter the increase in norepinephrine or glucagon elicited by hypoglycemia. These results indicate that during hypoglycemia, central excitatory amino acids contribute to the modulation of the glucoregulatory response through activation of NMDA receptors, resulting in stimulation of the sympathoadrenal and hypothalamic-pituitary adrenal axis. This mechanism appears to play an important role in the sustained elevation in hepatic glucose production during hypoglycemia.