Determining the importance of temporal regulation of the blood-brain barrier

项目摘要/摘要 生物钟在组织中普遍存在，调节许多生物功能，它们的 不对准或干扰会造成有害的健康后果。虽然是分子 生物钟的基础已经得到了很好的研究，生物钟在组织特异性生物中的作用 人们对血脑屏障(BBB)等功能知之甚少。BBB是一种 血管系统和大脑之间的接口，两者都保护大脑免受外周侮辱 并允许内源分子的运输。我们最近的研究发现，昼夜节律 CLOCK调节ATPase结合盒转运体介导的异种生物从脑中流出。 本研究旨在探讨血脑屏障昼夜节律调节的机制。 配体，它与睡眠和进食等行为的相关性，以及它在 发炎的状态。我认为，除了外来生物的外流外，生物钟在 血脑屏障调节来自大脑的颗粒的内吞作用(目标1)。基于已发布的和初步的 数据，我假设BBB时钟通过转运体调节来影响行为 内源性配体(目标2和3)。此外，我将询问炎症诱导的影响 通过内毒素或睡眠剥夺对血脑屏障节律的影响(目标4)。为了实现这些目标， 我将结合使用分子分析(定量聚合酶链式反应，细胞内离子测量， 代谢组学)、功能生物分析(内吞作用、异种生物渗透性)和行为 化验(睡眠、进食)。该项目的成功完成将在以下方面取得重要进展 了解血脑屏障的生物钟和行为。第一，了解分子 血脑屏障时钟的机制将进一步定义时钟在将血液输送到大脑中的作用 沟通。其次，它将为行为监管提供新的见解。第三，它将 鉴定新的内源性促进睡眠的化合物。理解时间门控 血脑屏障及其在病理生理条件下对行为的影响对发育具有重要意义 改善人类健康的干预措施。

Project Summary/Abstract Circadian clocks are ubiquitous in tissues, regulate many biological functions, and their misalignment or disruption contributes to deleterious health consequences. Although molecular underpinning of the circadian clock is well-studied, the role of the clock in tissue-specific biological functions, such as that of the blood-brain barrier (BBB), is poorly understood. The BBB is an interface between the vasculature and the brain that both protects the brain from peripheral insults and allows transports of endogenous molecules. Our recent work has found that the circadian clock regulates ATPase binding cassette transporter-mediated xenobiotic efflux from the brain. This proposal examines the mechanisms of BBB circadian clock regulation of endogenous ligands, its relevance to behaviors such as sleep and feeding, and its perturbation under conditions of inflammation. I propose that in addition to xenobiotic efflux, the circadian clock in the BBB regulates endocytosis of particles from brain (Aim 1). Based on published and preliminary data, I hypothesize that the BBB clock influences behavior through transporter-regulated endogenous ligands (Aim 2 and 3). Further, I will interrogate the effect of inflammation induced by either endotoxin or sleep deprivation on the rhythms of the BBB (Aim 4). To pursue these aims, I will use a combination of molecular assays (qPCR, intracellular ion measurements, metabolomics), functional bioassays (endocytosis, xenobiotic permeability) and behavioral assays (sleep, feeding). Successful completion of this project will offer important advances in understanding both the BBB circadian clock and behavior. First, understanding molecular mechanism of the BBB clock will further define the role of the clock in gating blood to brain communication. Second, it will provide new insights into the regulation of behavior. Third, it will identify novel endogenous sleep-promoting compounds. Understanding the temporal gating of the BBB and its effects on behavior under pathophysiologic conditions is important for developing interventions to improve human health.