Biology of Reproduction, Vol 25, 1159-1168, Copyright © 1981 by Society for the Study of Reproduction

Hormonally Responsive Areas of the Reproductive System of the Male Guinea Pig. III. Presence of Cytoplasmic Estrogen Receptors

¹ Department of Obstetrics and Gynecology, Department of Biochemistry, and Department of Anatomy, Vanderbilt University School of Medicine, Nashville, Tennessee 37232

We have previously shown that the administration of estrogens to intact adult male guinea pigs resulted in a variety of morphological changes in their accessory sex organs (the seminal vesicles, prostate, common ejaculatory chamber, coagulating gland, and epididymis). To ascertain if a biochemical basis existed for a direct action of estrogens on these tissues, we examined them for the presence of estrogen receptors. We determined that cytosol prepared from the accessory sex glands contains a macromolecular binding component for estradiol-17. This component sediments as an 8S species on 5-20% sucrose gradients under low (0.01 M KCl) ionic strength conditions and as a 4S species under high (0.4 M KCl) ionic strength conditions. Time-course studies indicate that binding equilibrium is achieved in 2 h; dissociation of [³H] estradiol from the binding protein is very slow (t>42 h). The cytoplasmic binder is highly specific for estrogens; the relative affinities of various estrogens for the protein were estradiol = 1, estrone = 0.72, diethylstilbestrol 0.30, and estriol = 0.17. Progesterone, testosterone, and 5-dihydrotestosterone, even at a 1000-fold excess, caused less than 10% inhibition of [³H] estradiol binding to the protein. The binder present in cytosol prepared from the accessory sex organs (excluding the epididymis) exhibited an equilibrium dissociation constant of 0.3 nM, and there were 30 fmoles of binding sites per mg of cytosol protein. The cytoplasmic estrogen binder exhibits the characteristics usually attributed to receptors and is clearly different from a 4S, nonspecific, rapidly dissociating binder that is present in plasma. On the basis of steroid specificity, the cytoplasmic estrogen binder is distinct from the androgen receptor that is present in male reproductive tissues.

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ACKNOWLEDGMENTS This research was supported by NIH grants HD 11706, HD 08295, HD 05797 (Population Center), and RR 05424 (BRSG). The authors wish to thank Ms. Esther Thomas for her excellent technical assistance and Ms. Vera Henley for typing the manuscript.