Male rats rendered diabetic by IV streptozotocin (65 mg/kg body weight) were treated with exogenous insulin or testosterone. Charcoal-coated dextran and polyacrylemide gel electrophoresis techniques were employed in studying the characteristics of androgen (R1881) binding to prostate cytosol protein. In comparison with normal (N) rats, the replacement therapy of diabetic (D) animals with insulin (D + I) or testosterone (D + T) was able to restore epididymal weight (N = 0.40 +/- 0.04 g; D = 0.18 +/0 0.02 g; D + I = 0.42 +/- 0.05 g; D + T = 0.40 +/0 0.06 g) and total prostate weight (N = 0.24 +/- 0.02 g; D = 0.15 +/- 0.02 g; D + I = 0.24 +/- 0.05 g; D + T = 0.35 +/- 0.06 h). Testicular endogenous content of testosterone was restored after insulin treatment (N = 154 +/- 13 ng/testis; D = 41 +/- 5 ng/testis; D + I = 142 +/- 9 ng/testis), and significant improvements of serum testosterone levels were also achieved (N = 540 +/- 64 ng/100 ml; D = 238 +/- 37 ng/100 ml; D + I = 358 +/- 18 ng/100 ml). Prostate cytosol of streptozotocin-diabetic rats had strongly lowered capacity for 3H-R1881 binding compared with controls (94 and 12 fmol/mg protein, respectively). Testosterone treatment produced a 3.3-fold improvement of this lowered value, whereas the increment seen with insulin was less (1.5-fold). It is emphasized that some of the improvements caused by insulin replacement therapy in diabetic animals are due to the partial restoration of testosterone secretion. Thus, the combined actions of insulin and testosterone (instead of insulin alone) seem to be of major importance in the maintenance and regulation of accessory sex glands function.