In vivo tracking of bioluminescent markers of circadian rhythms in behaving animals

项目摘要 昼夜节律是雕刻生理学的基础，它们的排列对 最佳健康状态。我们对细胞的基础以及细胞的功能有了深刻的了解 视交叉上核(SCN)的中央昼夜节律时钟，但我们还不知道 SCN与各种外设振荡器相互作用，以及这些外设时钟如何整合信息 从SCN-和非SCN-输入，以保持内部对齐以及持续24小时循环。 在这里，我们将研究两个外围振荡器，肝脏和皮肤。我们将开发一个动物模型，用于 研究这些外周时钟在昼夜节律系统中的作用，以确定它们的携带 能力。 这一提议的中心假设是，外周时钟保留了一些有节奏的 当SCN中央时钟受损时的功能，可能包括光夹带。长的- 这项研究的学期目标是了解整个生物钟的相互作用。 哺乳动物的身体，并最终发现新的工具来减轻昼夜节律失调的影响 关于健康。 我们的研究将首先旨在表征我们的新皮肤生物钟报告系K14CRE； DBPKI/+小鼠。我们将使用IVIS成像、分析和测试来更全面地描述我们的新鼠系 组织学以严格确定记录的生物发光的来源。第二，我们将致力于 在有或没有大脑中枢的小鼠体内测试光携带皮肤和肝脏的昼夜节律 时钟功能。这一目标将使用长期使用的方法测试皮肤和肝脏的夹带性 来自活体动物的昼夜生物发光报告，首先在完好的小鼠中，然后在有 视交叉上核(SCN)消融。第三，我们将确定这些外周组织是否显示出 比中央时钟的夹带范围更广。我们将测试基因昼夜节律的能力 K14Cre；DBPKI/+和Alb Cre的表达；DBPKI/+小鼠进入不同周期长度(“T”)的周期 周期“)，以评估夹带的范围，与运动活动的范围相比较，使用 完整和SCN损伤的小鼠。 这些研究的完成将极大地改变我们对哺乳动物的理解 昼夜节律系统，包括潜在的感光外周时钟。这项研究将是 由不同的、积极参与的本科生女性进行，并有系统的指导，以鼓励 继续从事生物医学科学的工作。

Project Summary Circadian rhythms are fundamental in sculpting physiology, and their alignment is essential for optimal health. We have gained deep understanding of the cellular basis as well as the function of the central circadian clock of the suprachiasmatic nuclei (SCN), but we don’t yet understand how the SCN interacts with varied peripheral oscillators, and how these peripheral clocks integrate information from SCN- and non-SCN-inputs to maintain internal alignment as well as entrainment to 24h cycles. Here we will study two peripheral oscillators, the liver and skin. We will develop an animal model for study of the role of these peripheral clocks within the circadian system, to determine their entrainment capabilities. The central hypothesis of this proposal is that peripheral clocks retain some rhythmic capabilities when the SCN central clock is impaired, possibly including photic entrainment. The long- term goal of this research is to understand the inter-dynamics of circadian clocks throughout the mammalian body and, ultimately, discover new tools to lessen the impact of circadian misalignment on health. Our research will first aim to characterize our new skin circadian clock reporter line K14 Cre; DBPKI/+ mice. We will more fully characterize our new mouse line using IVIS imaging, assays, and histology to rigorously determine sources of the bioluminescence recorded. Second, we will aim to test photic entrainment of skin and liver circadian rhythms in vivo in mice with or without brain central clock function. This aim will test the entrainment of skin and liver using methods for long-term circadian bioluminescent reporting from living animals, first in intact mice and then in mice with suprachiasmatic nuclei (SCN) ablation. Third, we will determine if these peripheral tissues show a wider range of entrainment than the central clock. We will test the ability for circadian rhythms of gene expression of K14 Cre; DBPKI/+ and Alb Cre; DBPKI/+ mice to entrain to cycles of varied cycle lengths (“T cycles”), to assess the range of entrainment, as compared with that of locomotor activity, using both intact and SCN-lesion mice. Completion of these studies will dramatically alter our understanding of the mammalian circadian system to include potentially photoreceptive peripheral clocks. The research will be conducted by diverse and engaged undergraduate women, with structured mentoring to encourage continued careers in biomedical science.