Specific binding of [3H]dexamethasone to cytosol and the activation of bound hormone-receptor complexes were studied in the kidney of immature (3-week) and mature (26-week) Long-Evans male rats. 2. The concentration of specific binding sites was significantly higher (25%) in the kidney of immature rats as compared with mature, while dissociation constants (Kd) remain unaltered at both ages. 3. Heat activation (25 degrees C for 45 min) significantly enhanced the binding of [3H]dexamethasone-receptor complexes to DNA-cellulose and purified nuclei at both ages to the same extent. Cross-mixing experiments (i.e. binding of activated cytosol from mature rats to nuclei of immature and vice versa) gave similar results to the non-mixed groups. 4. Ca2+ activation (0 degree C for 45 min with 20 mM Ca2+) also enhanced the nuclear and DNA-cellulose binding at both ages but to a greater magnitude in immature rats. 5. Differences in the number of specific binding sites and some of the physicochemical properties of kidney glucocorticoid receptors presented here between immature and mature rats may underlie the functional changes in tissue response with age.
摘要
在未成熟(3周龄)和成熟(26周龄)的Long-Evans雄性大鼠肾脏中,研究了[3H]地塞米松与胞浆的特异性结合以及结合的激素-受体复合物的激活情况。2. 与成熟大鼠相比,未成熟大鼠肾脏中特异性结合位点的浓度显著更高(25%),而两个年龄段的解离常数(Kd)保持不变。3. 热激活(25℃ 45分钟)在两个年龄段均显著增强了[3H]地塞米松-受体复合物与DNA-纤维素和纯化细胞核的结合,且程度相同。交叉混合实验(即将成熟大鼠的活化胞浆与未成熟大鼠的细胞核结合,反之亦然)得到了与未混合组相似的结果。4. Ca2+激活(0℃ 45分钟,加入20 mM Ca2+)在两个年龄段也增强了细胞核和DNA-纤维素的结合,但在未成熟大鼠中增强幅度更大。5. 此处呈现的未成熟和成熟大鼠肾脏糖皮质激素受体特异性结合位点数量以及一些物理化学性质的差异,可能是组织反应随年龄发生功能变化的基础。