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

行为动物昼夜节律生物发光标记的体内追踪

• 批准号: 10730688
• 负责人: MARY E HARRINGTON
• 金额: $ 39.63万
• 依托单位: SMITH COLLEGE
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-09-15 至 2026-09-17
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10730688
• 关键词: Ablation; Animal Model; Animals; Biological Assay; Bioluminescence; Brain; Cardiovascular system; Circadian Rhythms; Circadian desynchrony; Data; Gene Expression; Goals; Health; Histology; Impairment; Intervention; Laboratory mice; Length; Lesion; Light; Liver; Measures; Mentors; Metabolic; Methods; Motor Activity; Mus; Opsin; Periodicity; Peripheral; Physiology; Reporter; Reporting; Research; Role; Science; Skin; Source; Structure; Study models; System; Testing; Tissues; Woman; career; circadian; circadian pacemaker; improved; in vivo; in vivo imaging system; light effects; suprachiasmatic nucleus; tool; undergraduate student

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.

项目概要 昼夜节律是塑造生理学的基础，它们的协调对于塑造生理学至关重要 我们对细胞基础和功能有了深入的了解。 视交叉上核（SCN）的中央生物钟，但我们还不了解 SCN 与各种外设振荡器交互，以及这些外设时钟如何集成信息 来自 SCN 和非 SCN 输入以保持内部对齐以及 24 小时循环的夹带。 在这里，我们将研究两种外周振荡器，肝脏和皮肤，我们将开发一个动物模型。 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。 该提案的中心假设是外围时钟保留了一些节奏 SCN 中央时钟受损时的功能，可能包括光夹带。 这项研究的长期目标是了解整个昼夜节律时钟的相互作用 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。 我们的研究首先旨在描述我们新的皮肤生物钟报告系列 K14 Cre 的特征； 我们将使用 IVIS 成像、化验分析和方法更全面地表征我们的新小鼠品系。 其次，我们的目标是通过组织学严格确定记录的生物发光的来源。 在有或没有脑中枢的小鼠体内测试皮肤和肝脏昼夜节律的光夹带 此目的将使用长期方法测试皮肤和肝脏的夹带。 来自活体动物的昼夜节律生物发光报告，首先是在完整的小鼠中，然后是在有缺陷的小鼠中 第三，我们将确定这些周围组织是否显示出视交叉上核（SCN）消融。 比中央时钟更广泛的夹带范围我们将测试基因的昼夜节律能力。 K14 Cre 的表达；DBPKI/+ 和 DBPKI/+ 小鼠诱导不同周期长度的周期（“T”） 周期”），以评估夹带范围，与运动活动相比，使用两者 完整小鼠和 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 该研究将包括潜在的感光外围时钟的昼夜节律系统。 由多元化且积极参与的本科女性进行，并提供结构化指导以鼓励 继续从事生物医学科学事业。