Department of Biophysics and Biochemistry, Graduate School of Science, University of Tokyo, Tokyo 113-0033, Japan.
Proc Natl Acad Sci U S A. 2011 Mar 22;108(12):4864-9. doi: 10.1073/pnas.1015959108. Epub 2011 Mar 7.
The circadian clock is phase-delayed or -advanced by light when given at early or late subjective night, respectively. Despite the importance of the time-of-day-dependent phase responses to light, the underlying molecular mechanism is poorly understood. Here, we performed a comprehensive analysis of light-inducible genes in the chicken pineal gland, which consists of light-sensitive clock cells representing a prototype of the clock system. Light stimulated expression of 62 genes and 40 ESTs by >2.5-fold, among which genes responsive to the heat shock and endoplasmic reticulum stress as well as their regulatory transcription factors heat shock factor (HSF)1, HSF2, and X-box-binding protein 1 (XBP1) were strongly activated when a light pulse was given at late subjective night. In contrast, the light pulse at early subjective night caused prominent induction of E4bp4, a key regulator in the phase-delaying mechanism of the pineal clock, along with activation of a large group of cholesterol biosynthetic genes that are targets of sterol regulatory element-binding protein (SREBP) transcription factor. We found that the light pulse stimulated proteolytic formation of active SREBP-1 that, in turn, transactivated E4bp4 expression, linking SREBP with the light-input pathway of the pineal clock. As an output of light activation of cholesterol biosynthetic genes, we found light-stimulated pineal production of a neurosteroid, 7α-hydroxypregnenolone, demonstrating a unique endocrine function of the pineal gland. Intracerebroventricular injection of 7α-hydroxypregnenolone activated locomotor activities of chicks. Our study on the genome-wide gene expression analysis revealed time-of-day-dependent light activation of signaling pathways and provided molecular connection between gene expression and behavior through neurosteroid release from the pineal gland.
生物钟在主观夜晚的早期或晚期分别接收到光时,会出现相位延迟或提前。尽管光对时间依赖性相位反应的重要性不言而喻,但这种现象的潜在分子机制仍知之甚少。在这里,我们对鸡松果体中的光诱导基因进行了全面分析,鸡松果体由代表生物钟系统原型的感光时钟细胞组成。光照刺激了 62 个基因和 40 个 EST 的表达,其表达水平增加了 2.5 倍以上,其中对热休克和内质网应激反应的基因以及其调节转录因子热休克因子(HSF)1、HSF2 和 X 盒结合蛋白 1(XBP1)在主观夜晚晚期接收到光脉冲时被强烈激活。相比之下,在主观夜晚的早期接收到光脉冲会引起 E4bp4 的明显诱导,E4bp4 是松果体时钟相位延迟机制的关键调节剂,同时还会激活一大组胆固醇生物合成基因,这些基因是固醇调节元件结合蛋白(SREBP)转录因子的靶基因。我们发现,光脉冲刺激了活性 SREBP-1 的蛋白水解形成,而 SREBP-1 又反式激活了 E4bp4 的表达,将 SREBP 与松果体时钟的光输入途径联系起来。作为胆固醇生物合成基因被光激活的产物,我们发现光刺激了松果体中神经甾体 7α-羟孕酮的产生,证明了松果体的独特内分泌功能。脑室注射 7α-羟孕酮可激活小鸡的运动活动。我们对全基因组基因表达分析的研究揭示了生物钟信号通路随时间变化的光激活,通过松果体中神经甾体的释放,为基因表达和行为之间提供了分子联系。