Mak P, Callard G V
J Steroid Biochem. 1985 Mar;22(3):355-61. doi: 10.1016/0022-4731(85)90438-8.
Although the hamster is frequently used as an experimental animal for studying reproductive neuroendocrinology and sex behavior, estrogen receptors (ER) in the central nervous system have not been fully characterized. Using Sephadex LH-20 gel filtration and DNA-cellulose affinity chromatography, estrogen binding macromolecules having the physicochemical properties of classical ER were identified in cytosolic and nuclear extracts of brain tissues. These receptors exhibited high affinity for estradiol (Kd = 10(-9) M), limited capacity (30-50 fmol/g tissue), and estrogen specificity; however, competition studies indicate that brain and uterine ER have different binding kinetics. The neuroanatomic distribution of ER was similar in males and females with highest levels in the limbic brain and consistently low levels in remaining forebrain and mid/hindbrain. No sex differences in receptor number or other binding parameters were evident. Sucrose gradient centrifugation showed that cytosolic ER sedimented in the 7-8S region of a 5-20% linear gradient (no salt), whereas nuclear ER had a sedimentation coefficient of 5S under high ionic strength. On DNA-cellulose affinity columns, these receptors had an elution maximum of 0.18 M NaCl. After a single injection of estradiol, nuclear ER increased and cytosolic ER were depleted. The lower estradiol binding affinity and receptor levels in hamster brain as compared to the rat are consistent with observed species differences in neural sensitivity to estrogen. We expect these data in hamsters, a markedly photosensitive species, to provide a basis for future studies examining the role of receptors in mediating the effects of day-length on steroid dependent feedback and behavioral responses.
尽管仓鼠常被用作研究生殖神经内分泌学和性行为的实验动物,但中枢神经系统中的雌激素受体(ER)尚未得到充分表征。利用葡聚糖LH - 20凝胶过滤和DNA - 纤维素亲和色谱法,在脑组织的胞质和核提取物中鉴定出具有经典ER物理化学性质的雌激素结合大分子。这些受体对雌二醇表现出高亲和力(Kd = 10^(-9) M)、有限的容量(30 - 50 fmol/g组织)和雌激素特异性;然而,竞争研究表明,脑和子宫的ER具有不同的结合动力学。ER的神经解剖分布在雄性和雌性中相似,在边缘脑区水平最高,而在前脑其余部分和中脑/后脑水平始终较低。在受体数量或其他结合参数上没有明显的性别差异。蔗糖梯度离心显示,胞质ER在5 - 20%线性梯度(无盐)的7 - 8S区域沉降,而核ER在高离子强度下的沉降系数为5S。在DNA - 纤维素亲和柱上,这些受体的最大洗脱浓度为0.18 M NaCl。单次注射雌二醇后,核ER增加,胞质ER减少。与大鼠相比,仓鼠脑中较低的雌二醇结合亲和力和受体水平与观察到的物种对雌激素神经敏感性的差异一致。我们期望这些来自明显光敏感物种仓鼠的数据,能为未来研究受体在介导日长对类固醇依赖性反馈和行为反应影响中的作用提供基础。