Division of Pharmacology, Ohio State University, Columbus, OH, 43210, USA.
Department of Neuroscience, Ohio State University, Columbus, OH, 43210, USA.
Eur J Neurosci. 2018 Apr;47(7):845-857. doi: 10.1111/ejn.13850. Epub 2018 Feb 19.
The circadian timing system influences the functional properties of most, if not all, physiological processes. Central to the mammalian timing system is the suprachiasmatic nucleus (SCN) of the hypothalamus. The SCN functions as a 'master clock' that sets the phasing of ancillary circadian oscillator populations found throughout the body. Further, via an entraining input from the retina, the SCN ensures that the clock oscillators are synchronized to the daily light/dark cycle. A critical component of the SCN timing and entrainment systems is the p44/42 mitogen-activated protein kinase (ERK/MAPK) pathway. Here, we examined the expression and function of phosphoprotein-enriched in astrocytes (PEA-15), an ERK scaffold protein that serves as a key regulator of MAPK signaling. A combination of immunolabeling and Western blotting approaches revealed high levels of PEA-15 within the SCN. PEA-15 expression was enriched in distinct subpopulations of SCN neurons, including arginine vasopressin (AVP)-positive neurons of the SCN shell region. Further, expression profiling detected a significant circadian oscillation in PEA-15 expression within the SCN. Brief photic stimulation during the early subjective night led to a significant increase in PEA-15 phosphorylation, an event that can trigger ERK/PEA-15 dissociation. Consistent with this, co-immunoprecipitation assays revealed that PEA-15 is directly bound to ERK in the SCN and that photic stimulation leads to their dissociation. Finally, we show that PEA-15 regulates ERK/MAPK-dependent activation of the core clock gene period1. Together, these data raise the prospect that PEA-15 functions as a key regulator of the SCN timing system.
昼夜节律计时系统影响着大多数(如果不是全部的话)生理过程的功能特性。哺乳动物计时系统的核心是下丘脑的视交叉上核(SCN)。SCN 作为一个“主时钟”,设定了遍布全身的辅助昼夜节律振荡器群体的时相。此外,通过来自视网膜的传入信号的同步作用,SCN 确保了时钟振荡器与日常的光/暗循环同步。SCN 计时和同步输入系统的一个关键组成部分是 p44/42 丝裂原活化蛋白激酶(ERK/MAPK)通路。在这里,我们研究了富含星形胶质细胞的磷酸蛋白(PEA-15)的表达和功能,PEA-15 是一种 ERK 支架蛋白,是 MAPK 信号转导的关键调节因子。免疫标记和 Western blot 分析方法的结合揭示了 SCN 中高水平的 PEA-15。PEA-15 在 SCN 神经元的不同亚群中表达丰富,包括 SCN 壳区的精氨酸加压素(AVP)阳性神经元。此外,表达谱分析检测到 SCN 中 PEA-15 表达存在显著的昼夜节律性波动。在主观夜晚早期进行短暂的光刺激会导致 PEA-15 磷酸化显著增加,这一事件可以触发 ERK/PEA-15 解离。与此一致,共免疫沉淀实验表明 PEA-15 直接与 SCN 中的 ERK 结合,并且光刺激会导致它们的解离。最后,我们表明 PEA-15 调节 ERK/MAPK 依赖性核心时钟基因 period1 的激活。总之,这些数据表明 PEA-15 作为 SCN 计时系统的关键调节因子发挥作用。