In vivo tools for analyzing interstitial fluid flow

用于分析间质液流动的体内工具

• 批准号: 9751865
• 负责人: Nozomi Nishimura
• 金额: $ 20.5万
• 依托单位: CORNELL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-08-01 至 2021-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9751865
• 关键词: Address; Affect; Alzheimer' s Disease; Amyloid beta-Protein; Anatomy; Architecture; Astrocytes; Biological; Biological Process; Blood; Blood Vessels; Brain; Cells; Characteristics; Clinical; Cognition; Conflict (Psychology); Disease; Drug Delivery Systems; Dura Mater; Edema; Engineering; Equilibrium; Excision; Extracellular Space; Flushing; Genes; Grant; Hippocampus (Brain); Image; Imaging technology; In Situ; Injections; Intercellular Fluid; Ischemia; Label; Lasers; Link; Literature; Lymph; Lymphatic; Lymphatic System; Lymphatic vessel; Magnetic Resonance Imaging; Maintenance; Measurement; Measures; Medical; Memory; Meningeal; Meninges; Methods; Motion; Mus; Optics; Organ; Paper; Pathway interactions; Peptides; Play; Process; Production; Proteins; Reporting; Research Design; Resolution; Role; Route; Scanning; Signal Transduction; Site; Sleep; Sleep disturbances; Source; Speed; Stroke; Structure; Surface; System; Textbooks; Time; Tissues; Tracer; Transgenic Mice; Viral Vector; Waste Products; Water; Whole Organism; Work; abeta accumulation; arteriole; brain cell; brain tissue; brain volume; cranium; cytokine; experimental study; extracellular; fluid flow; fluorescence imaging; image reconstruction; imaging modality; in vivo; injured; innovation; interest; interstitial; ionic balance; migration; multiphoton imaging; multiphoton microscopy; novel; pressure; solute; stroke model; theories; three photon microscopy; tool; tumor; two photon microscopy; two-photon; wasting; water channel

Interstitial fluid (ISF) flow has many functions including the maintenance of ionic balances, flushing of wastes and providing a route for migration of both cell signals and cells. Recently, the use of multiphoton microscopy, which enables in vivo studies with cellular resolution, has resulted in novel findings, especially in the brain, about dynamics and anatomy involved in ISF flow. Notably, ISF flow may be critical in dealing with protein accumulation in Alzheimer's disease and is regulated by sleep. Although much progress has been made, there remains controversy about some of the fundamentals regarding ISF flow. Much of this may be due to complications in the experimental methods. Studies to date require the injection of tracers which can be imaged by multiphoton microscopy or other imaging methods such as MRI. However, the process of injection of the tracers may itself affect the flow due to the delicate balance of pressures within the brain. In addition, injected tracers do not mimic the origin of proteins, wastes and cytokines made by the brain. Studies are limited to superficial sites accessible by current generation imaging technologies. This proposal generates optical and biological tools that address these short comings using in vivo multiphoton microscopy. First, in a new way to generate tracers in situ, cells within the tissue of interests will be transduced so that they secrete fluorescent proteins into the extracellular space. This will be used to resolve existing controversies about the route of ISF flow within the brain. Second, the newly discovered brain lymphatics are thought to link to the peri or paravacular spaces that serve as conduits for ISF. This work will use the new secrete tracers to answer whether and how these lymphatic channels link to the these spaces. This fluid flow may be altered in different conditions, so this will be studied in normal function mimicking sleep and waking, as well as with a stroke model. Third, three-photon microscopy now enables much deeper imaging than traditional two-photon microscopy. This enables imaging of anatomy previously not accessible. This work will study ISF flow in the hippocampus, a critical brain structure in memory and cognition, that seems to be particularly vulnerable to disruptions to ISF. This work will establish novel tools that can enable new experiments to address ISF in many systems.

间质流体(ISF)流动具有多种功能，包括维持离子 平衡，废物的冲洗，并提供一条移动的路线，细胞信号和 细胞。最近，多光子显微镜的使用，使活体研究能够 细胞分辨率，导致了关于动力学的新发现，特别是在大脑中 以及参与ISF血流的解剖学。值得注意的是，ISF流可能在处理 蛋白质在阿尔茨海默病中积累，并受到睡眠的调节。虽然很多 已经取得了进展，但对一些基本面仍然存在争议 关于ISF流程。这在很大程度上可能是由于实验中的并发症造成的 方法：研究方法。到目前为止的研究需要注射示踪剂，可以通过 多光子显微镜或其他成像方法，如核磁共振。然而，这一过程 由于微妙的平衡，示踪剂的注入本身可能会影响流动 大脑内的压力。此外，注入的示踪剂不能模拟 由大脑产生的蛋白质、废物和细胞因子。研究仅限于肤浅的场地。 可由当代成像技术访问。这一提议产生了光学 以及利用体内多光子解决这些缺陷的生物工具 显微镜。首先，在一种新的原位生成示踪剂的方法中，组织内的细胞 兴趣将被转导，以便它们将荧光蛋白分泌到 细胞外空间。这将被用来解决关于路线的现有争议 ISF在大脑中流动。其次，新发现的脑淋巴管被认为是 链接到充当ISF管道的腔周或腔旁间隙。这项工作将使用 新的分泌示踪剂用于回答这些淋巴通道是否以及如何与 这些空间。这种流体流动在不同的条件下可能会发生变化，因此将对此进行研究 在正常功能下模仿睡眠和清醒，以及中风模型。第三, 与传统的双光子显微镜相比，三光子显微镜现在可以实现更深入的成像 显微镜。这使得以前无法访问的解剖成像成为可能。这项工作将 研究海马体的ISF流动，海马体是记忆和认知的关键大脑结构， 这似乎特别容易受到ISF中断的影响。这项工作将确立 新的工具可以使新的实验在许多系统中解决ISF问题。