Deep Analysis of Brain Chemistry at Enhanced Spatial and Temporal Resolution using Microscale Sampling and Analysis

使用微尺度采样和分析以增强的时空分辨率深入分析脑化学

基本信息

批准号：
10515445
负责人：
ROBERT T KENNEDY
金额：
$ 140.4万
依托单位：
UNIVERSITY OF MICHIGAN AT ANN ARBOR
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-08-01 至 2025-07-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10515445
关键词：
Address Affect Area BRAIN initiative Behavioral Model Biological Assay Blood Circulation Brain Brain Chemistry Brain Diseases Caliber Cell Maintenance Cell physiology Cells Chemicals Chronic Complement Complex Concentration measurement Coupled Coupling Disadvantaged Disease Energy Supply Extracellular Space Glutamates Glutamine Goals Immunoassay In Situ Length Lipids Liquid substance Location Mass Spectrum Analysis Measurement Measures Mental disorders Metabolic Metabolic Pathway Methods Microdialysis Modernization Monitor Neuroglia Neuronal Differentiation Neurons Neurotransmitters Organ Physiological Production Proteins Protocols documentation Psyche structure Reporting Resolution Sampling Societies Source Specificity Spectrometry, Mass, Electrospray Ionization Stable Isotope Labeling System Techniques Technology Testing Time Work analytical method base behavioral phenotyping brain cell capillary liquid chromatography design experimental study gamma-Aminobutyric Acid implanted sensor improved in vivo in vivo monitoring instrument instrumentation mental state metabolome metabolomics nanolitre nervous system disorder neurochemistry neurotransmitter release novel operation prototype public health relevance receptor sensor small molecule stable isotope temporal measurement tool ultra high resolution

项目摘要

The objective of this project is to develop new bioanalytical methods for exploring brain chemistry dynamics in vivo. Monitoring the concentration dynamics of neurochemicals in vivo is vital for studying brain function, diseases, and treatments. A versatile approach for in vivo monitoring of brain chemistry is to couple sampling methods, such as microdialysis, to analytical measurements. Primary advantages of the sampling methods relative to other tools, such as sensors, are the ability to perform continuous measurements over time, measure a wide variety of chemicals, and potential for metabolic tracing; however, these potential advantages are underdeveloped. The disadvantages of the method are the poor temporal and spatial resolution. Temporal resolution is limited by the time required to collect enough samples for analysis while spatial resolution is limited by the probe size. In this project, we will develop technology to overcome these limitations. Further, the strengths will be developed and the unique capabilities to address the needs of the Brain Initiative. To facilitate long-term measurements at the high spatial resolution, novel microfabricated sampling probes will be developed that allow measurements at the scale of just a few cells. To improve temporal resolution, instrumentation for coupling fast assays based on mass spectrometry and proximity immunoassay to the new sampling probes will be developed. These methods will allow continuous measurement of neurotransmitters, metabolites, and proteins in real-time with 1 s temporal resolution. A second aim is to develop metabolomics, i.e. measurement of the full complement of lipids and metabolites present, for samples collected from the brain extracellular space in vivo. For this aim, ultra-high-resolution capillary liquid chromatography coupled to mass spectrometry will be developed to analyze the metabolome present in probe samples. This method will allow the chemical milieu of the brain extracellular space to be characterized in unprecedented detail over time and at specific brain locations. In this way, it will be possible to discover chemicals and metabolic pathways that govern different mental and disease states. A third aim will be to use stable-isotope tracing to detect the neuronal pool of glutamate and GABA, the primary neurotransmitters in the brain. These chemicals have multiple cellular sources so previous studies have been limited in understanding if detected changes in concentration were due to neuronal function of other cell activities. This capability will allow a new understanding of the dynamics of these neurotransmitters, which are involved in a myriad of brain functions.

该项目的目的是开发新的生物分析方法来探索体内脑化学动力学。监测体内神经化学物质的浓度动力学对于研究脑功能，疾病和治疗至关重要。用于体内监测脑化学的多功能方法是将采样方法（例如微透析）与分析测量相结合。采样方法相对于其他工具（例如传感器）的主要优点是能够随着时间的推移进行连续测量，测量多种化学物质以及代谢追踪的潜力。但是，这些潜在的优势不发达。该方法的缺点是时间和空间分辨率差。时间分辨率受到收集足够样品进行分析所需的时间的限制，而空间分辨率受探针大小的限制。在这个项目中，我们将开发技术来克服这些局限性。此外，将发展优势以及满足大脑计划需求的独特能力。为了促进高空间分辨率的长期测量，将开发出新的微型抽样探针，以仅在几个细胞的尺度上进行测量。为了提高时间分辨率，将开发基于质谱和接近新采样探针的接近免疫测定的快速测定的仪器。这些方法将允许通过1 s时间分辨率实时测量神经递质，代谢产物和蛋白质。第二个目的是开发代谢组学，即对存在的脂质和代谢物的完整补体的测量，用于从体内收集的样品。为此，将开发出与质谱法结合的超高分辨率毛细管液相色谱法，以分析探针样品中存在的代谢组。这种方法将使大脑的化学环境随着时间的流逝和特定的大脑位置的前所未有的细节来表征。这样，可以发现控制不同心理和疾病状态的化学和代谢途径。第三个目标是使用稳定的异位跟踪来检测谷氨酸和GABA的神经元库，GABA是大脑中的主要神经递质。这些化学物质具有多种细胞来源，因此以前的研究在理解浓度的变化是否是由于其他细胞活性的神经元功能引起的。这种能力将使对这些神经递质的动力学有了新的了解，这些神经递质涉及无数的大脑功能。