Morrell J I, McGinty J F, Pfaff D W
Neuroendocrinology. 1985 Nov;41(5):417-26. doi: 10.1159/000124212.
We used the combined steroid autoradiography-immunocytochemical method to determine whether estradiol- or dexamethasone-concentrating cells contain endogenous opioid peptides. Ovariectomized-adrenalectomized female rats were given highly radioactive doses of 3H-estradiol or 3H-dexamethasone, then sacrificed to demonstrate nuclear steroid binding. Autoradiograms were prepared, exposed for 2-12 months, photodeveloped, and fixed; immunocytochemistry was carried out on the same sections using antibodies to beta-endorphin or dynorphin A (1-17). In the medial basal hypothalamus, many estradiol- and some dexamethasone-concentrating neurons were found intermingled with beta-endorphin or dynorphin-immunoreactive neurons. Of the beta-endorphin-immunoreactive neurons in the medial basal hypothalamus, 4% concentrated estradiol in their nuclei. In addition, a subset of beta-endorphin-immunoreactive cells in the anterior pituitary concentrated estradiol in their nuclei. Although none of the beta-endorphin-immunoreactive neurons in the medial basal hypothalamus concentrated dexamethasone in their nuclei, many of the beta-endorphin-immunoreactive cells in the anterior pituitary did. Of the dynorphin-immunoreactive neurons in the medial basal hypothalamus, 10% concentrated estradiol in their nuclei. These data are consistent with the hypothesis of a genomic effect of estradiol on a particular subset of medial basal hypothalamic neurons that produce endogenous opioid peptides.
我们采用类固醇放射自显影术与免疫细胞化学相结合的方法，以确定雌二醇或地塞米松浓缩细胞是否含有内源性阿片肽。对卵巢切除及肾上腺切除的雌性大鼠给予高放射性剂量的³H-雌二醇或³H-地塞米松，然后处死以显示核类固醇结合情况。制备放射自显影片，曝光2至12个月，显影并固定；在同一切片上使用抗β-内啡肽或强啡肽A（1-17）抗体进行免疫细胞化学检测。在内侧基底部下丘脑，发现许多雌二醇浓缩神经元以及一些地塞米松浓缩神经元与β-内啡肽或强啡肽免疫反应性神经元相互交织。在内侧基底部下丘脑的β-内啡肽免疫反应性神经元中，有4%的细胞核浓缩了雌二醇。此外，垂体前叶中一部分β-内啡肽免疫反应性细胞的细胞核也浓缩了雌二醇。虽然内侧基底部下丘脑的β-内啡肽免疫反应性神经元中没有一个细胞核浓缩地塞米松，但垂体前叶中有许多β-内啡肽免疫反应性细胞的细胞核浓缩了地塞米松。在内侧基底部下丘脑的强啡肽免疫反应性神经元中，有10%的细胞核浓缩了雌二醇。这些数据与雌二醇对内侧基底部下丘脑产生内源性阿片肽的特定神经元亚群具有基因组效应的假说相符。
