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

项目摘要 昼夜节律是塑造生理学的基础，它们的排列对于 最佳健康我们已经对细胞基础以及细胞的功能有了深入的了解。 视交叉上核（SCN）的中央昼夜节律钟，但我们还不清楚 SCN与各种外围振荡器相互作用，以及这些外围时钟如何整合信息 从SCN-和非SCN-输入，以保持内部对齐以及夹带到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。我们将建立一个动物模型， 研究这些外围时钟在昼夜节律系统中的作用，以确定它们的夹带 能力的 这一提议的核心假设是，外围时钟保留了一些节奏， 当SCN中央时钟受损时，可能包括光夹带。很长的- 本研究的长期目标是了解整个过程中生物钟的相互动态 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新的皮肤生物钟报告细胞系K14 Cre; DBPKI/+小鼠。我们将使用IVIS成像、测定和免疫分析更全面地表征我们的新小鼠系， 组织学以严格确定记录的生物发光的来源。第二，我们的目标是 在有或没有脑中枢神经系统小鼠中测试皮肤和肝脏昼夜节律的体内光夹带 时钟功能这一目标将测试的夹带皮肤和肝脏使用的方法，为长期 来自活体动物的昼夜节律生物发光报告，首先在完整小鼠中，然后在具有 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 比中央时钟更宽的夹带范围。我们将测试基因的昼夜节律的能力， 表达K14 Cre; DBPKI/+和Alb Cre; DBPKI/+小鼠以诱导不同周期长度的周期（“T 周期”），以评估夹带的范围，与自发活动的范围相比，使用 完整和SCN损伤小鼠。 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，包括潜在的感光外围时钟。这项研究将 由多元化和参与的本科女性进行，有结构化的指导，以鼓励 继续从事生物医学科学。

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.