Cryptochrome 2 expression level is critical for adrenocorticotropin stimulation of cortisol production in the capuchin monkey adrenal.
Cryptochrome 2 expression level is critical for adrenocorticotropin stimulation of cortisol production in the capuchin monkey adrenal.
复制标题
隐花色素 2 的表达水平对于促肾上腺皮质激素刺激卷尾猴肾上腺皮质醇的产生至关重要。
作者：
C. Torres;L. Abarzúa;F. Valenzuela;N. Méndez;H. Richter;G. Valenzuela;M. Seron
响应ACTH及时产生糖皮质激素是生存所必需的，通过协调能量的摄入和消耗，并作为对抗压力的稳态调节剂。肾上腺皮质醇对ACTH的反应是时钟时间依赖性的，这表明肾上腺皮质中的内在昼夜节律振荡器有助于调节对ACTH的反应。生物钟基因的表达已被报道在肾上腺皮质的几个物种。然而，目前还没有报道说明肾上腺钟蛋白对皮质醇对ACTH反应的潜在影响。在这里，我们探讨了时钟蛋白隐花色素2（CCR 2）敲低是否会改变灵长类动物肾上腺对ACTH的反应。将来自成年卷尾猴（n = 5）的肾上腺外植体与转染载体（对照）或与两种不同的Cry 2反义和正义探针预孵育6小时，然后在单独的培养基（无ACTH）或与100 nm ACTH孵育48小时。在控制和感觉条件下，促肾上腺皮质激素增加皮质醇的产生，而抑制β 2抑制促肾上腺皮质激素刺激的皮质醇的产生。类固醇生成酶类固醇生成急性调节蛋白和3 β-羟基类固醇脱氢酶在48小时的孵育增加ACTH在控制外植体和抑制Cry 2敲低。此外，我们发现Cry 2敲低降低了时钟基因脑和肌肉芳香烃受体核转位样蛋白（Bmal 1）在mRNA和蛋白水平的表达。总之，这些结果强烈支持，时钟蛋白β 2参与的机制，促肾上腺皮质激素增加类固醇生成急性调节蛋白和3 β-羟基类固醇脱氢酶的表达。因此，肾上腺生物钟的组成部分的适当表达水平是必要的适当的皮质醇对ACTH的反应。
Timely production of glucocorticoid hormones in response to ACTH is essential for survival by coordinating energy intake and expenditure and acting as homeostatic regulators against stress. Adrenal cortisol response to ACTH is clock time dependent, suggesting that an intrinsic circadian oscillator in the adrenal cortex contributes to modulate the response to ACTH. Circadian clock gene expression has been reported in the adrenal cortex of several species. However, there are no reports accounting for potential involvement of adrenal clock proteins on cortisol response to ACTH. Here we explored whether the clock protein cryptochrome 2 (CRY2) knockdown modifies the adrenal response to ACTH in a primate. Adrenal gland explants from adult capuchin monkey (n = 5) were preincubated for 6 h with transfection vehicle (control) or with two different Cry2 antisense and sense probes followed by 48 h incubation in medium alone (no ACTH) or with 100 nm ACTH. Under control and sense conditions, ACTH increased cortisol production, whereas CRY2 suppression inhibited ACTH-stimulated cortisol production. Expression of the steroidogenic enzymes steroidogenic acute regulatory protein and 3beta-hydroxysteroid dehydrogenase at 48 h of incubation was increased by ACTH in control explants and suppressed by Cry2 knockdown. Additionally, we found that Cry2 knockdown decreased the expression of the clock gene brain and muscle aryl hydrocarbon receptor nuclear translocator-like protein (Bmal1) at the mRNA and protein levels. Altogether these results strongly support that the clock protein CRY2 is involved in the mechanism by which ACTH increases the expression of steroidogenic acute regulatory protein and 3beta-hydroxysteroid dehydrogenase. Thus, adequate expression levels of components of the adrenal circadian clock are required for an appropriate cortisol response to ACTH.