New Findings r What is the central question of this study?Thyroid dysfunction can have a major impact on pancreatic function. The influence of hyperthyroidism on insulin secretion remains controversial, and the precise mechanism of its effect has not yet been elucidated. r What is the main finding and its importance?The results of this study demonstrate that hyperthyroidism leads to impaired insulin secretion. It appears that the defect in insulin secretion in the hyperthyroid state probably reflects a summation of different alterations, including decreased sensitivity of ATP-sensitive K + and L-type Ca 2+ channels of the β-cells and reduced β-cell mass.To clarify the mechanism underlying the effect of thyroid hormone excess on pancreatic insulin secretion and abnormal glucose tolerance induced by hyperthyroidism, we investigated the effect of hyperthyroidism on the pancreatic β-cell mass and two key components of the insulin secretory pathway, ATP-sensitive K + (K ATP ) and L-type Ca 2+ channels. In control and levothyroxine-treated hyperthyroid rats, an intraperitoneal glucose tolerance test was performed, and the insulin secretion and content of the isolated islets were assayed. In order to determine the effect of hyperthyroidism on K ATP and L-type Ca 2+ channels, isolated islets were exposed to specific pharmacological agents, including glibenclamide (K ATP channel blocker), diazoxide (K ATP channel opener) and nifedipine (L-type Ca 2+ channel blocker). Histomorphometric changes and histochemistry of the islet in both groups were compared. Our data indicated that plasma glucose and insulin concentrations during the intraperitoneal glucose tolerance test in the hyperthyroid group were, respectively, higher and lower than in the control group. Insulin secretion and content of the hyperthyroid islets were reduced. The response of hyperthyroid islets to glibenclamide, diazoxide and nifedipine and the percentage change in insulin secretion were lower than those of the control islets. Despite the increase in weight and total volume of the pancreas, the volume of the islets and the total number of insulin-positive cells in hyperthyroid rats were reduced. Our data indicated that reduced insulin secretion in the hyperthyroid group might arise from reduced β-cell mass and an abnormality in some parts of the insulin secretory pathway, including K ATP and L-type Ca 2+ channel function.