Partial characterization and ontogeny of renal cytosolic glucocorticoid receptors in mouse kidney.
The glucocorticoid receptor (GR) was partially characterized in mouse renal cytosol. A sensitive and reproducible [3H]dexamethasone binding assay suitable for use with small quantities of cytosolic protein, was developed. Studies defined the optimal equilibrium binding conditions, metabolism of [3H]dexamethasone in adult renal cytosol, specificity of binding of the GR, and molecular weight of the GR-[3H]dexamethasone complex by gel filtration chromatography. The assay was subsequently used to measure the renal GR during different stages of foetal and postnatal development, as well as in glomerular and renal tubular preparations from adult mice. An almost linear increase in GR occurred from day 13 to day 18 of gestation with levels rising from 100 to 201 fmol/mg cytosol protein; this was followed by a sharp rise in receptor concentration just after birth to 343 fmol/mg cytosol protein. Adult levels, 410-433 fmol/mg cytosol protein, were reached by 2 weeks after birth. The equilibrium dissociation constants (Kd) of the [3H]dexamethasone-receptor complex were similar in adult and in embryonic cytosols (range, 2.8-11.8 nM; mean +/- SD = 6.5 +/- 2.9 nM). Specific binding was assessed to be 3- to 5-fold greater in tubular than in glomerular preparations. These data on the localization and ontogeny of GR during murine metanephric development provide a basis for study of glucocorticoid-mediated effects on various models of congenital and acquired renal disease.
D000450:Aldosterone / Q000378:metabolism; D000818:Animals; D003345:Corticosterone / Q000378:metabolism; D003600:Cytosol / Q000032:analysis; D003907:Dexamethasone / Q000378:metabolism; D004229:Dithiothreitol / Q000494:pharmacology; D004958:Estradiol / Q000378:metabolism; D005260:Female; D006854:Hydrocortisone / Q000378:metabolism; D007668:Kidney / Q000254:growth & development* / Q000378:metabolism; D007678:Kidney Glomerulus / Q000378:metabolism; D007684:Kidney Tubules / Q000378:metabolism; D051379:Mice; D008970:Molecular Weight; D008982:Molybdenum / Q000494:pharmacology; D011247:Pregnancy; D011374:Progesterone / Q000378:metabolism; D011965:Receptors, Glucocorticoid / Q000378:metabolism*; D013379:Substrate Specificity; D013739:Testosterone / Q000378:metabolism
D011965:Receptors, Glucocorticoid; C044659:molybdate; D013739:Testosterone; D000450:Aldosterone; D011374:Progesterone; D004958:Estradiol; D003907:Dexamethasone; D008982:Molybdenum; D004229:Dithiothreitol; D003345:Corticosterone; D006854:Hydrocortisone