Shih Meng-Chun, Hsu Nai-Chi, Huang Chen-Che, Wu Tzong-Shoon, Lai Pao-Yen, Chung Bon-Chu
Institute of Molecular Biology, 48, Academia Sinica, Nankang, Taipei 115, Taiwan.
Mol Endocrinol. 2008 Apr;22(4):915-23. doi: 10.1210/me.2007-0222. Epub 2008 Jan 3.
Steroids are synthesized mainly from the adrenal glands catalyzed by steroidogenic enzymes; the expression of these enzymes is controlled by transcription factor steroidogenic factor-1 (SF-1; NR5A1). To understand the physiological effect of genetic changes on steroid secretion, we used Cre-LoxP and gene targeting technology to mutate the binding sequence for SF-1 (SF-1 response element) on the promoter of the mouse Cyp11a1 gene, which encodes a critical enzyme for steroid biosynthesis. The resulting Cyp11a1 L/L mice expressed about 7-fold less cytochrome P450 side-chain cleavage enzyme (CYP11A1) in the adrenal and testis but expressed normal amounts of CYP11A1 in the placenta and ovary. This tissue-specific reduction of gene expression did not affect basal steroid secretion but attenuated the circadian rhythm of glucocorticoid secretion. These mice also failed to induce glucocorticoid secretion in response to stress, leading to retention of CD4+CD8+ double-positive thymocytes. Unlike complete Cyp11a1 disruption, which causes neonatal death, promoter mutation did not decrease life span and caused no defect in reproduction. Thus, CYP11A1 appears in normal mice to be expressed above the minimal required level, providing a large capacity for use in response to stress. Mutation of the SF-1 response element of Cyp11a1 results in reduced stress response due to decreased adrenal CYP11A1 expression and insufficient stress-induced glucocorticoids secretion.
类固醇主要由肾上腺在类固醇生成酶的催化下合成;这些酶的表达受转录因子类固醇生成因子-1(SF-1;NR5A1)控制。为了解基因变化对类固醇分泌的生理影响,我们使用Cre-LoxP和基因靶向技术,使小鼠Cyp11a1基因启动子上SF-1的结合序列(SF-1反应元件)发生突变,该基因编码类固醇生物合成的关键酶。产生的Cyp11a1 L/L小鼠肾上腺和睾丸中细胞色素P450侧链裂解酶(CYP11A1)的表达量约减少7倍,但胎盘和卵巢中CYP11A1的表达量正常。这种基因表达的组织特异性降低并不影响基础类固醇分泌,但减弱了糖皮质激素分泌的昼夜节律。这些小鼠在应激时也无法诱导糖皮质激素分泌,导致CD4+CD8+双阳性胸腺细胞滞留。与导致新生儿死亡的完全Cyp11a1破坏不同,启动子突变并未缩短寿命,也未导致生殖缺陷。因此,在正常小鼠中,CYP11A1的表达似乎高于最低所需水平,为应对应激提供了很大的能力。Cyp11a1的SF-1反应元件突变导致应激反应降低,原因是肾上腺CYP11A1表达减少以及应激诱导的糖皮质激素分泌不足。