Abstract Dopaminergic systems have been implicated in diabetes and obesity. Notwithstanding, the most basic relationship between dopamine and plasma insulin as well as glucose levels yet remains unknown. The present experiments were designed to investigate the effects of acute hyperinsulinemia on basal dopamine levels in the nucleus accumbens of the rat under chloral hydrate anesthesia using acute microdialysis in combination with the hyperinsulinemic–glycemic clamping procedure. In Experiment 1, each rat was infused with one of the three concentrations of insulin (2.4, 4.8, or 9.6 mU/kg per min) while plasma glucose levels were maintained at euglycemia (∼ 5.5 mmol/L). Dopamine, dihydroxyphenylacetic acid and homovanillic acid were not significantly different from baseline during either the clamp or post-clamp periods for all insulin concentrations. In Experiment 2, rats were infused with the highest concentration of insulin (9.6 mU/kg per min) and plasma glucose levels were maintained at either hypoglycemia (∼ 3 mmol/L) or hyperglycemia (∼ 14 mmol/L). Dopamine was elevated at 100 min (+ 113% above basal levels) and 120 min (+ 117%) in the hypoglycemic condition and at 120 min (+ 121%) in the hyperglycemic condition. In the hyperglycemic post-clamp period, homovanillic acid was decreased below basal levels (approximately − 32%). These results together suggest that short-term blood glucose deviations coupled with acute hyperinsulinemia affect the mesoaccumbens dopamine system.