In vivo tools for analyzing interstitial fluid flow

用于分析间质液流动的体内工具

基本信息

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
项目类别：
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&apos 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

项目摘要

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.

间质液 (ISF) 流动具有多种功能，包括维持离子平衡、冲洗废物并提供细胞信号和迁移的途径细胞。最近，多光子显微镜的使用使得体内研究成为可能细胞分辨率，产生了关于动力学的新发现，特别是在大脑中以及 ISF 流涉及的解剖学。值得注意的是，ISF 流在处理阿尔茨海默病中的蛋白质积累，并受睡眠调节。虽然很多已经取得了进展，但一些基本原则仍存在争议关于 ISF 流量。这在很大程度上可能是由于实验中的复杂性造成的方法。迄今为止的研究需要注射示踪剂，示踪剂可以通过以下方式成像多光子显微镜或其他成像方法，例如 MRI。然而，该过程由于示踪剂的微妙平衡，示踪剂的注射本身可能会影响流量脑内的压力。此外，注射的示踪剂并不模仿大脑产生的蛋白质、废物和细胞因子。研究仅限于表面场所通过当前一代成像技术即可实现。该提案产生光学以及使用体内多光子解决这些缺点的生物工具显微镜。首先，以一种原位产生示踪剂的新方法，组织内的细胞兴趣将被转换，以便它们分泌荧光蛋白到细胞外空间。这将用于解决有关路线的现有争议 ISF 在大脑内流动。其次，新发现的脑淋巴管被认为连接到作为 ISF 导管的周围或血管旁空间。本作品将使用新的分泌示踪剂来回答这些淋巴通道是否以及如何与这些空间。这种流体流动可能会在不同条件下改变，因此将对此进行研究模拟睡眠和清醒的正常功能以及中风模型。第三，三光子显微镜现在能够比传统的双光子显微镜更深层次地成像显微镜。这使得以前无法获得的解剖结构成像成为可能。这项工作将研究海马体中的 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 干扰的影响。这项工作将建立新颖的工具可以使新的实验能够解决许多系统中的 ISF 问题。