Faccioli G, Minelli G, Bolelli G F
Dipartimento di Biologia, Universita di Bologna, Italy.
Arch Ital Biol. 1989 Jun;127(3):173-86.
A polyclonal antiserum against androgens, i.e., testosterone, 5alpha-dihydrotestosterone and androstenedione, was tested to reveal target neurons of endogenous androgens in the hypothalamus of both intact and castrated male rats. Paraffin sections of hypothalamus and testis were immunostained by using Avidin-Biotin Complex method and 3-3' diaminobenzidine to visualize the immunoperoxidase complex. Conventional control experiments for method and antiserum specificity were performed. The antiserum proved to be specific for androgens, i.e., testosterone, 5alpha-dihydrotestosterone and androstenedione. The nuclear labeling observed in tissues stained by this procedure is consistent with the hypothesis that the labeled neurons contained DHT, which is the main testosterone metabolite active in the cell nucleus. The antiserum was effective in staining not only the hypothalamic neurons of intact males with normal serum levels of testosterone but also the hypothalamic neuron of castrated males with very low serum levels of testosterone. Evidence is presented indicating that the immunostaining technique represents a more specific and sensitive method to identify target neurons of endogenous androgens than autoradiography.
检测了一种针对雄激素（即睾酮、5α-双氢睾酮和雄烯二酮）的多克隆抗血清，以揭示完整和阉割雄性大鼠下丘脑中内源性雄激素的靶神经元。采用抗生物素蛋白-生物素复合物法和3,3'-二氨基联苯胺对下丘脑和睾丸的石蜡切片进行免疫染色，以观察免疫过氧化物酶复合物。进行了方法和抗血清特异性的常规对照实验。该抗血清被证明对雄激素（即睾酮、5α-双氢睾酮和雄烯二酮）具有特异性。通过该程序染色的组织中观察到的核标记与以下假设一致，即标记的神经元含有双氢睾酮，它是在细胞核中具有活性的主要睾酮代谢产物。该抗血清不仅能有效染色睾酮血清水平正常的完整雄性大鼠的下丘脑神经元，还能染色睾酮血清水平极低的阉割雄性大鼠的下丘脑神经元。有证据表明，与放射自显影相比，免疫染色技术是一种更特异、更灵敏的鉴定内源性雄激素靶神经元的方法。
