Demonstration of Partial Characterization of Cytosol Receptors for Testosterone

Jan-Ake Gustafsson, Åke Pousette

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Androgen uptake was investigated in several peripheral organs after administration of [1,2,6,7-3H]testosterone to castrated male rats. The animals were killed after 30 min, the organs were taken out, and the radioactivity was determined after tissue combustion. A relatively high accumulation of androgen was found in pancreas, adrenals, spleen, thigh muscle, kidneys, and liver in addition to the classical androgen target organs coagulation glands, seminal vesicles, prostate, preputial glands, and harderian glands. In a second series of experiments, nuclear and cytosol fractions were prepared from prostate, seminal vesicles, coagulation glands, preputial glands, spleen, submaxillary glands, kidneys, and pancreas from castrated male rats given [l,2,6,7-3H]testosterone, and these fractions were then characterized by thin-layer and radio-gas chromatography with respect to their patterns of labeled steroids. Only prostate and seminal vesicles were found to contain significant amounts of nuclear 5α-[3H]dihydrotestosterone. The major nuclear androgen was [3H]testosterone that was the only detectable labeled steroid in coagulation glands, preputial glands, and spleen and that constituted 70% or more of the nuclear radioactivity in seminal vesicles, submaxillary glands, kidneys, and pancreas. These results indicate that testosterone itself may be the predominant active androgen principle in vivo in most androgen target organs and that conversion to 5α-dihydrotestosterone is generally not a prerequisite for androgen activity. Using an ultrasensitive micromodification of isoelectric focusing (cf. M. Katsumata and A. S. Goldman (1974), Biochem. Biophys. Ada 359, 112. It was possible to show that cytosol from kidney,- submaxillary gland, thigh muscle, and levator ani muscle and nuclei from kidney and submaxillary gland contained androgen-binding proteins with p/'s in the region 4.6-5.1 ("4.6-5.1 Complex"). This complex also formed in vitro after incubation of [l,2,6,7-3H]testosterone with cytosol from kidney and submaxillary gland. [l,2,6,7-3H]Testosterone was bound with high affinity to receptor proteins in cytosol from both kidney, submaxillary gland, and thigh muscle with dissociation constants of 5.0 × 10-12 M (kidney), 3.3 × 10-11 M and 4.1 × 10-10 M (two types of binding sites, submaxillary gland), 2.4 × 10-12 M (thigh muscle) and 1.9 × 10-12 M (levator ani muscle). The number of binding sites was in all cases between 1 and 20 fmol/mg of protein. On the basis of these results the hypothesis is presented that a common class of testosterone receptors is present in most organs and that these receptors can be detected both in vivo and in vitro provided methods sensitive enough are utilized.

Original languageEnglish (US)
Pages (from-to)3094-3101
Number of pages8
Issue number14
StatePublished - Jul 1 1975

ASJC Scopus subject areas

  • Biochemistry


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