Technology for functional study of cells and circuits in large postmortem brains ex vivo

离体大型死后大脑细胞和电路功能研究技术

基本信息

批准号：
9928247
负责人：
NENAD SESTAN
金额：
$ 15.56万
依托单位：
YALE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-08-01 至 2021-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9928247
关键词：
Address Algorithms Anatomy Animal Feed Animal Model Animals Area Automation Autopsy Axon Biological Brain Brain region Cardiovascular system Cell physiology Cells Cellular Morphology Cephalic Cerebrum Chemicals Communities Complex Custom Disease Electrocorticogram Electrolytes Electrophysiology (science)Engineering Family suidae Food production Formulation Functional Imaging Functional disorder Gases Genetic Glucose Goals Grant Harvest Health Hippocampus (Brain)Histologic Histology Homeostasis Hour Human Investigation Kinetics Label Lead Maintenance Mammals Mechanics Mental disorders Metabolism Methodology Microcirculation Modeling Molecular Molecular Analysis Molecular Structure Monitor Names Neurons Neurosciences Nutrient Operative Surgical Procedures Output Perfusion Physiologic pulse Physiological Positron-Emission Tomography Pulsatile Flow Quality Control Research Research Personnel Resolution Resuscitation Scanning Source Structure Synapses System Techniques Technology Temperature Time Tissue Viability Tissues Tracer Translations Validation X-Ray Computed Tomography Zoology base brain tissue cell type density experimental study functional outcomes imaging capabilities imaging modality imaging study improved in vivo multimodality neocortical nervous system disorder neural circuit neurotechnology new technology non-genetic novel novel strategies patch clamp preservation pressure prototype relating to nervous system restoration sensor tissue culture tool user-friendly vector

项目摘要

PROJECT SUMMARY The mammalian brain is arguably the most complex biological structure. Investigating cellular functions and mapping neural connections in the brain are critical tasks to better understand the brain in health and disease. This is particularly challenging in vivo due to the inherent limitations in experimental latitude and simultaneous access to multiple brain regions within the same animal. These shortcomings hinder multimodal interrogation of multi-synaptic circuits and mesoscale connectomics. Of particular importance, these experimental inadequacies grow in proportion to the complexity of the brain and cranial anatomy, impeding translation to larger mammals. This grant addresses these tasks by optimizing and validating a first-in-class neurotechnology called BrainEx for the restoration of molecular and cellular functions of the postmortem large mammalian brain under ex vivo, normothermic conditions. We specifically propose to continue optimizing BrainEx in porcine brains, while validating the efficacy of the BrainEx system as a new experimental platform for electrophysiological, connectomic, and imaging studies in the fully isolated, intact, and functional large mammalian brain. There are four major distinguishing aspects of this application: (1) implementation of novel approaches developed to restore cerebral macro- and microcirculation and extend cellular viability of the postmortem brain under normothermic conditions such that researchers can (2) simultaneously trace connections and characterize cellular function and morphology by chemical and vector-based techniques across myriad brain regions, including areas inaccessible to in vivo surgical approaches; (3) investigate multisynaptic long-range circuitry and cortical network electrical activity; and (4) perform functional PET and CT imaging studies in the ex vivo large mammalian brain. This methodology represents a new tool for more thorough investigation of the structure and function of complex circuits and the cells within them. Wide distribution of this technology will grant investigators experimental advantages across species not afforded by tissue culture or in vivo approaches.

项目概要哺乳动物的大脑可以说是最复杂的生物结构。研究细胞功能和绘制大脑中的神经连接是更好地了解健康和疾病状态下的大脑的关键任务。由于实验范围和同步的固有限制，这在体内尤其具有挑战性。访问同一动物的多个大脑区域。这些缺点阻碍了多模式审讯多突触回路和中尺度连接组学。特别重要的是，这些实验的不足之处其生长与大脑和颅骨解剖结构的复杂性成正比，阻碍了向大型哺乳动物的转化。这笔赠款通过优化和验证名为 BrainEx 的一流神经技术来解决这些任务在离体条件下恢复死后大型哺乳动物大脑的分子和细胞功能，常温条件。我们特别建议继续优化猪脑中的 BrainEx，同时验证 BrainEx 系统作为电生理学新实验平台的功效，对完全分离、完整且有功能的大型哺乳动物大脑进行连接组学和成像研究。有该应用程序的四个主要区别方面：（1）实施开发的新方法恢复大脑宏观和微循环，并延长死后大脑的细胞活力常温条件下，研究人员可以 (2) 同时追踪连接并表征通过基于化学和载体的技术跨越无数大脑区域的细胞功能和形态，包括体内手术无法到达的区域； (3) 研究多突触远程电路和皮质网络电活动； (4) 在离体大样本中进行功能性 PET 和 CT 成像研究哺乳动物的大脑。该方法代表了一种新工具，可以更彻底地研究结构和复杂电路及其内部细胞的功能。这项技术的广泛分布将为研究人员提供帮助组织培养或体内方法无法提供跨物种的实验优势。