This study was conducted to determine whether there is a increase in responsiveness to the activating effects of testosterone on male reproductive behavior during puberty in male golden hamsters and whether responsiveness to behavioral actions of testosterone is correlated with the ability of testosterone to upregulate brain androgen receptor immunoreactivity (AR-ir). Sexually naive male hamsters were castrated at 21 or 42 days of age and implanted subcutaneously with a pellet containing 0, 2.5, or 5 mg of testosterone. One week later, males were given a 10-min mating test with a receptive female. Animals were euthanized 1 hr after the behavioral test, and blood samples and brains were collected. Plasma testosterone levels were equivalent in prepubertal and adult males that had been administered the same dose of testosterone. However, adult males exhibited more mounts, intromissions, and ejaculations than prepubertal males, demonstrating that postpubertal males are more responsive than prepubertal males to the effects of testosterone on sexual behavior. In both age groups, testosterone increased the number of AR-ir cells per unit area in several brain regions involved in male sexual behavior, including the medial preoptic nucleus (MPN), medial amygdala, posteromedial bed nucleus of the stria terminalis, and magnocellular preoptic nucleus (MPNmag). Surprisingly, testosterone increased AR-ir in the latter three regions to a greater extent in prepubertal males than in adults. Thus, prepubertal males are more responsive to the effects of testosterone on AR-ir in these regions. In a separate experiment, a pubertal increase in the number of AR-ir cells per unit area was found in both the MPN and MPNmag of intact male hamsters. These results indicate that a testosterone-dependent increase in brain AR during puberty may be necessary, but is not sufficient, to induce an increase in behavioral responsiveness to testosterone.