This report deals with a comparative study of the uptake of radioactivity in the genital, neural and other tissues of intact male, castrate male and female rats after constant infusion of radiochemically pure 1,2-3H-testosterone. Castration has been found to have a marked effect in enhancing the uptake of radioactivity in the different tissues particularly liver and kidney which have a metabolic and excretory role. Prostate and seminal vesicles, on the other hand, did not show any difference in uptake in intact and castrate rats. The uptake of testosterone and its metabolites in the prostate of intact animals was 3.46 × 10−9 m. In experiments on subcellular localization of radioactivity after constant infusion of 1,2-3H-testosterone, it was found that prostate and seminal vesicle had heavy localisation in the nuclear and 105 000 × g soluble fraction while the major localisation of radioactivity in the case of nontarget tissues like liver, intestine, muscle etc. was in the soluble fraction. Castration caused a higher uptake of radioactivity in the anterior pituitary gland, anterior and middle hypothalamus also, which could be explained on the basis of the negative feedback of the hormone. An interesting feature of the uptake of testosterone and its metabolites in the female rat was the high uptake in the anterior pituitary gland, and the various parts of the hypothalamus. These findings are discussed in light of information available on the action and feedback of sex hormones.