The in vivo effects of colchicine on the subcellular distribution of insulin receptors have been studied in insulin-injected rats and in control animals. Colchicine (0.1 mg/100 g or 10 mg/100 g body weight, i.v.) did not affect the ability of plasma membranes and Golgi fractions of control rats to bind insulin. As previously reported (Desbuquois et al., 1982), the injection of native insulin (8 nmol, i.v.) caused a 50% decrease in the insulin binding activity of plasma membranes and a concomitant 50% increase in insulin binding to Golgi fractions. These changes occurred at 4 and 40 min after insulin injection but were no longer detectable at 3 h. Colchicine treatment did not affect the initial changes in the distribution of insulin receptors induced by insulin; however, in rats treated with the low dose of colchicine, insulin binding to plasma membranes at 3 h was not fully restored. Colchicine treatment did not alter the amount of acid-extractable insulin associated with Golgi fractions of insulin-injected rats. The time course of uptake of 125I-insulin was similar in plasma membranes, microsomal fraction and Golgi fractions of colchicine-treated (0.1 mg/100 g) and of untreated rats. These results suggest that colchicine does not interfere with the endocytosis of insulin receptors induced by their ligand and has little effect, if any, on the reinsertion of internalized receptors in the plasma membrane.