Estrogen Receptor in Rat Adrenal Gland

Gordon B. Cutler, Kevin M. Barnes, Mark A. Sauer, D. Lynn Loriaux

Research output: Contribution to journalArticle

53 Scopus citations

Abstract

Specific binding of [3H]estradiol by the cytoplasmic fraction of rat adrenal gland was studied at 0 C. The [3H]estradiol-binding component sedimented at 7 S on 5 to 20% sucrose gradients. Scatchard analysis of the cytosol binding reaction, measured by a charcoal-dextran assay, indicated a single class of estradiol binding sites of high affinity (Kd = 2.2 - 4.5 × 10-10 M) and limited number (5-22 fmol/mg cytosol protein) (n = 7). Competition studies indicated specificity of the binding reaction for the biologically active estrogens, estradiol, estrone, estriol, and diethylstilbestrol; binding of [3H]estradiol was unaffected by a 1000- fold excess of androgens, glucocorticoids, mineralocorticoids, or progesterone. The binding activity was temperature dependent (no binding was observed when incubations were performed at 25 C or above) and appeared to be by a protein since binding was destroyed by trypsin and sulfhydryl reagents, but not by ribonuclease or deoxyribonuclease. Kinetic studies indicated an apparent second order association rate constant of 3.3 × 10-5 M-1 sec-1 and a first order dissociation rate constant of 3.1 × 10-5 sec-1 at 0 C. Specific binding of [3H]estradiol by rat adrenal nuclei was also studied utilizing a nuclear exchange assay. One hour after estrogen injection in vivo, specific binding of [3H]estradiol by isolated adrenal nuclei in vitro increased more than 8-fold, suggesting nuclear translocation of the cytoplasmic adrenal estrogen receptor. Scatchard analysis of the specific nuclear binding indicated a single class of saturable high affinity sites with a dissociation equilibrium constant of 5 × 10-10 M. Since the specific [3H]estradiol-binding component of rat adrenal gland has the properties of a physiological estrogen receptor, the possibility of a direct effect of estrogens on rat adrenal cortical function is suggested.

Original languageEnglish (US)
Pages (from-to)252-257
Number of pages6
JournalEndocrinology
Volume102
Issue number1
DOIs
StatePublished - Jan 1978

ASJC Scopus subject areas

  • Endocrinology

Fingerprint Dive into the research topics of 'Estrogen Receptor in Rat Adrenal Gland'. Together they form a unique fingerprint.

  • Cite this

    Cutler, G. B., Barnes, K. M., Sauer, M. A., & Loriaux, D. L. (1978). Estrogen Receptor in Rat Adrenal Gland. Endocrinology, 102(1), 252-257. https://doi.org/10.1210/endo-102-1-252