Schlinger B A, Amur-Umarjee S, Campagnoni A T, Arnold A P
Department of Physiological Science, University of California, Los Angeles 90024, USA.
J Neuroendocrinol. 1995 Mar;7(3):187-92. doi: 10.1111/j.1365-2826.1995.tb00746.x.
Enzymes in the avian brain irreversibly catalyze the conversion of androgens into either active metabolites (aromatase and 5 alpha-reductase) or inactive metabolites (5 beta-reductase), 5 beta-reductase is thought to influence the formation of active metabolites by reducing the concentration of androgenic substrate available for these reactions. However, because these enzymes have different regional, cellular and subcellular distributions in brain, the traditional method to measure enzyme activity in brain homogenates may be inaccurate because of artificial mixing of enzymes. Recently, we have prepared primary cell cultures from the telencephalons of developing zebra finches. Cell cultures offer the advantage that enzyme activity can be measured in live cells in which the relative distribution of enzymes may more closely reflect that found in vivo. We have previously reported that aromatase is expressed at high levels in these cultures, and that it is active in both neurons and in glia. Here we report on the activities of 5 alpha- and 5 beta-reductase in these cell cultures. Along with aromatase, 5 beta-reductase was expressed at high levels in these mixed cell cultures, including cultures highly enriched in glia. This suggests that 5 beta-reductase is present in non-neuronal cells in brain, possibly co-localized with aromatase. However, despite the presence of 5 beta-reductase, aromatase was detected reliably in vitro even when the concentration of substrate was low. Thus, 5 beta-reductase does not prevent the synthesis of estrogen in the telencephalon of developing zebra finches. By contrast, 5 alpha-reductase activity was very low or absent in these cultures.(ABSTRACT TRUNCATED AT 250 WORDS)
鸟类大脑中的酶不可逆地催化雄激素转化为活性代谢物(芳香化酶和5α-还原酶)或无活性代谢物(5β-还原酶),5β-还原酶被认为通过降低这些反应可用的雄激素底物浓度来影响活性代谢物的形成。然而,由于这些酶在大脑中具有不同的区域、细胞和亚细胞分布,传统的测量脑匀浆中酶活性的方法可能不准确,因为酶会人为混合。最近,我们从发育中的斑胸草雀的端脑中制备了原代细胞培养物。细胞培养具有可以在活细胞中测量酶活性的优势,其中酶的相对分布可能更紧密地反映体内的情况。我们之前报道过这些培养物中芳香化酶高表达,并且它在神经元和神经胶质细胞中均有活性。在此我们报道这些细胞培养物中5α-和5β-还原酶的活性。与芳香化酶一起,5β-还原酶在这些混合细胞培养物中高表达,包括高度富集神经胶质细胞的培养物。这表明5β-还原酶存在于大脑的非神经元细胞中,可能与芳香化酶共定位。然而,尽管存在5β-还原酶,即使底物浓度较低时,体外仍能可靠地检测到芳香化酶。因此,5β-还原酶并不妨碍发育中的斑胸草雀端脑中雌激素的合成。相比之下,这些培养物中5α-还原酶活性非常低或不存在。(摘要截选至250词)