Warembourg M, Jolivet A, Milgrom E
INSERM U 156, Lille, France.
Brain Res. 1989 Feb 20;480(1-2):1-15. doi: 10.1016/0006-8993(89)91561-8.
Estrogen and progesterone interact in the regulation of various brain functions including mechanisms controlling gonadotropin secretion and female sexual behavior. Various methods have been used to map the regions where these hormones act and where specific receptors can be detected. However, it remains unknown if both steroids act on the same neuron or if there are neural populations responding to either one or the other of these hormones To answer this question, we used various immunocytochemical procedures to detect estrogen and progesterone receptors (ER-IR and PR-IR) on the same histological section, taking advantage of the fact that anti-ER monoclonal antibodies were raised from rats whereas anti-PR monoclonal antibodies were raised from mice. Initial experiments showed that the number of cells displaying ER and PR immunoreactivity changed with hormonal treatment. Prolonged treatment of ovariectomized guinea pigs with high doses of estradiol benzoate (EB) (10 or 15 micrograms/day for 4 or 6 days) increased the number of PR-positive cells in the preoptic area and the hypothalamus. Inversely, with this estrogen priming regimen the number of ER-positive cells decreased, as did the immunostaining intensity in their nuclei. With a lower dose of estrogen administered for a shorter period (2-microgram dose of EB for 2 days) the two receptor populations could be easily observed. Under these hormonal conditions, the sequential procedure revealed that the vast majority of cells containing ER-IR in the preoptic area (nucleus preopticus periventricularis, medialis) and the mediobasal hypothalamus (nucleus periventricularis, arcuatus, ventrolateral part of nucleus ventromedialis and premamillaris) showed PR immunoreactivity. This was true regardless of the order in which dual immunocytochemical staining was performed. There were, however, a few ER-positive/PR-negative cells in these regions. However, only ER-positive neurons were found in the n. interstitialis stria terminalis, the n. amygdaloideus medialis and the n. supraopticus. Thus, in guinea pigs receiving moderate doses of estrogen, all PR-positive cells contain ER-IR whereas there is a population of ER-positive cells which are devoid of PR-IR.
雌激素和孕酮在多种脑功能的调节中相互作用，这些脑功能包括控制促性腺激素分泌的机制以及雌性性行为。人们已采用各种方法来绘制这些激素作用的区域以及可检测到特定受体的区域。然而，这两种甾体激素是否作用于同一神经元，或者是否存在对其中一种或另一种激素有反应的神经群体，目前仍不清楚。为了回答这个问题，我们利用抗雌激素单克隆抗体是从小鼠制备而抗孕酮单克隆抗体是从大鼠制备这一事实，采用各种免疫细胞化学方法在同一组织切片上检测雌激素和孕酮受体（雌激素受体免疫反应性和孕酮受体免疫反应性）。初步实验表明，显示雌激素和孕酮免疫反应性的细胞数量会随激素处理而变化。用高剂量苯甲酸雌二醇（EB）（10或15微克/天，持续4或6天）对去卵巢豚鼠进行长期处理，会增加视前区和下丘脑孕酮阳性细胞的数量。相反，采用这种雌激素预处理方案，雌激素阳性细胞的数量会减少，其细胞核中的免疫染色强度也会降低。给予较低剂量的雌激素并处理较短时间（2微克剂量的EB，持续2天），可以很容易地观察到这两种受体群体。在这些激素条件下，连续的实验步骤表明，视前区（室周视前核、内侧）和下丘脑内侧基底部（室周核、弓状核、腹内侧核腹外侧部分和乳头前核）中绝大多数含有雌激素受体免疫反应性的细胞显示孕酮免疫反应性。无论进行双重免疫细胞化学染色的顺序如何，都是如此。然而，在这些区域有一些雌激素阳性/孕酮阴性细胞。不过，在终纹床核、内侧杏仁核和视上核中只发现了雌激素阳性神经元。因此，在接受中等剂量雌激素的豚鼠中，所有孕酮阳性细胞都含有雌激素受体免疫反应性，而存在一群缺乏孕酮受体免疫反应性的雌激素阳性细胞。
