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

• 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
• 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

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.

项目摘要 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， 绘制大脑中的神经连接是更好地了解大脑健康和疾病的关键任务。 这在体内是特别具有挑战性的，这是由于在实验范围和同时进行的实验中的固有限制。 进入同一动物的多个大脑区域。这些缺点阻碍了多模式询问， 多突触回路和中尺度连接组学。尤其重要的是，这些实验上的不足 与大脑和颅骨解剖结构的复杂性成比例增长，阻碍了向大型哺乳动物的转化。 这项资助通过优化和验证一种名为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脑的分子和细胞功能， 常温条件下。我们特别建议继续优化猪脑中的BrainEx， 验证BrainEx系统作为电生理学新实验平台的有效性， 在完全分离的、完整的和功能性的大型哺乳动物脑中的连接组学和成像研究。有 本申请的四个主要区别方面：（1）开发的新方法的实施， 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）同时跟踪连接和表征 细胞功能和形态通过化学和基于载体的技术跨越无数的大脑区域， 包括体内手术方法无法触及的区域;（3）研究多突触长程回路， 皮质网络电活动;和（4）在离体大的 哺乳动物的大脑这种方法是一种新的工具，可以更彻底地调查结构， 复杂电路和其中的细胞的功能。这项技术的广泛传播将使研究人员 组织培养或体内方法无法提供跨物种的实验优势。