Department of Neuroscience, Ohio State University, Columbus, OH 43210, USA.
Neuroscience. 2011 May 5;181:79-88. doi: 10.1016/j.neuroscience.2011.03.005. Epub 2011 Mar 5.
Circadian (24-h) rhythms influence virtually every aspect of mammalian physiology. The main rhythm generation center is located in the suprachiasmatic nucleus (SCN) of the hypothalamus, and work over the past several years has revealed that rhythmic gene transcription and post-translational processes are central to clock timing. In addition, rhythmic translation control has also been implicated in clock timing; however the precise cell signaling pathways that drive this process are not well known. Here we report that a key translation activation cascade, the mammalian target of rapamycin (mTOR) pathway, is under control of the circadian clock in the SCN. Using phosphorylated S6 ribosomal protein (pS6) as a marker of mTOR activity, we show that the mTOR cascade exhibits maximal activity during the subjective day, and minimal activity during the late subjective night. Importantly, expression of S6 was not altered as a function of circadian time. Rhythmic S6 phosphorylation was detected throughout the dorsoventral axis of the SCN, thus suggesting that rhythmic mTOR activity was not restricted to a subset of SCN neurons. Rather, rhythmic pS6 expression appeared to parallel the expression pattern of the clock gene period1 (per1). Using a transgenic per1 reporter gene mouse strain, we found a statistically significant cellular level correlation between pS6 and per1 gene expression over the circadian cycle. Further, photic stimulation triggered a coordinate upregulation of per1 and mTOR activation in a subset of SCN cells. Interestingly, this cellular level correlation between mTOR activity and per1 expression appears to be specific, since a similar expression profile for pS6 and per2 or c-FOS was not detected. Finally, we show that mTOR activity is downstream of the ERK/MAPK signal transduction pathway. Together these data reveal that mTOR pathway activity is under the control of the SCN clock, and suggests that mTOR signaling may contribute to distinct aspects of the molecular clock timing process.
昼夜节律(24 小时)影响着哺乳动物生理学的几乎所有方面。主要的节律生成中心位于下丘脑的视交叉上核(SCN),过去几年的工作表明,节律性基因转录和翻译后过程是时钟计时的核心。此外,节律性翻译控制也与时钟计时有关;然而,驱动这一过程的确切细胞信号通路尚不清楚。在这里,我们报告哺乳动物雷帕霉素靶蛋白(mTOR)途径的一个关键翻译激活级联反应受 SCN 生物钟的控制。我们使用磷酸化 S6 核糖体蛋白(pS6)作为 mTOR 活性的标志物,表明 mTOR 级联在主观白天表现出最大的活性,而在主观深夜表现出最小的活性。重要的是,S6 的表达不是作为昼夜节律时间的函数而改变的。节律性 S6 磷酸化在 SCN 的背腹轴上都有检测到,因此表明节律性 mTOR 活性不仅限于 SCN 神经元的一部分。相反,节律性 pS6 表达似乎与时钟基因 period1(per1)的表达模式平行。使用 per1 报告基因的转基因小鼠品系,我们发现 pS6 和 per1 基因表达在昼夜周期上存在统计学上显著的细胞水平相关性。此外,光刺激引发了 SCN 细胞中 per1 和 mTOR 激活的协调上调。有趣的是,mTOR 活性与 per1 表达之间的这种细胞水平相关性似乎是特异性的,因为没有检测到 pS6 和 per2 或 c-FOS 的相似表达谱。最后,我们表明 mTOR 活性是 ERK/MAPK 信号转导通路的下游。这些数据共同表明,mTOR 途径的活性受 SCN 时钟的控制,并表明 mTOR 信号可能有助于分子时钟计时过程的不同方面。