Vanson A, Arnold A P, Schlinger B A
Department of Psychology, University of California, Los Angeles 90024, USA.
Gen Comp Endocrinol. 1996 Jun;102(3):342-50. doi: 10.1006/gcen.1996.0077.
3 beta-hydroxysteroid dehydrogenase/delta 5 - delta 4 isomerase (3 beta-HSD) activity was measured in primary dissociated cell cultures prepared from telencephalons of developing zebra finches. 3 beta-HSD activity was confirmed after cultures were incubated with [7-3H]pregnenolone (Preg) or (1,2,6,7-3H-) dehydroepiandrosterone (DHEA) and 3H-progesterone (Prog) and 3H-androstenedione (AE) were detected in the medium. Product identity was confirmed by recrystallizations and by HPLC analysis. When DHEA was used as substrate, 3H-estradiol and 3H-estrone were also detected in the culture medium, presumably derived from the aromatization of 3H-AE or 3H-T produced from 3H-DHEA. To test this idea, cultures were incubated with 3H-DHEA together with radioinert AE or with fadrozole HCl, a potent and specific aromatase inhibitor. In the presence of radioinert AE, 3H-AE increased but metabolites of 3H-AE decreased in the media; in the presence of fadrozole, 3H-estrogens decreased but 3H-AE and its androgenic metabolite 3H-5 beta-androstanedione increased. These data demonstrate 3 beta-HSD activity in the songbird brain. The presence of Prog and estradiol in these cultures suggest that Preg and DHEA can potentially serve as substrates for the ultimate formation of active sex steroids in the songbird telencephalon.
在从发育中的斑胸草雀端脑制备的原代解离细胞培养物中测量了3β-羟基类固醇脱氢酶/δ5-δ4异构酶(3β-HSD)活性。在用[7-3H]孕烯醇酮(Preg)或(1,2,6,7-3H-)脱氢表雄酮(DHEA)孵育培养物后,确认了3β-HSD活性,并在培养基中检测到3H-孕酮(Prog)和3H-雄烯二酮(AE)。通过重结晶和高效液相色谱分析确认了产物的同一性。当使用DHEA作为底物时,在培养基中还检测到3H-雌二醇和3H-雌酮,推测它们来自3H-DHEA产生的3H-AE或3H-T的芳香化作用。为了验证这一想法,将培养物与3H-DHEA以及放射性惰性AE或强效特异性芳香化酶抑制剂盐酸法倔唑一起孵育。在存在放射性惰性AE的情况下,培养基中3H-AE增加但3H-AE的代谢产物减少;在存在法倔唑的情况下,3H-雌激素减少但3H-AE及其雄激素代谢产物3H-5β-雄烷二酮增加。这些数据证明了鸣禽脑中存在3β-HSD活性。这些培养物中Prog和雌二醇的存在表明,Preg和DHEA可能潜在地作为鸣禽端脑中活性性类固醇最终形成的底物。