BRAIN Initiative: Integrated Multimodal Analysis of Cell and Circuit-Specific Processes in Hippocampal Function

 描述(申请人提供)：大脑计划旨在了解大脑的空间、时间和化学性质。RFA-MH-15-225要求开发具有若干目标的新工具和技术，包括从单个细胞及其连接获得细胞类型和化学信息的方法。虽然存在许多使用特定探针来成像细胞亚群及其相互连接的成像方法，但该项目将创建一种化学信息丰富的方法，以促进新兴的受激拉曼散射显微镜(SRSM)技术的发展。SRSM提供活体脑片每个位置的振动光谱数据，因此可以跟踪动态的化学变化。拉曼光谱包含大量的化学信息，但这些数据是在复杂的重叠分子振动带中编码的。有了适当的训练集--从拉曼数据中获得，并将其与单个细胞的化学成分进行比较--一系列数学模型将被开发出来，创建无限的“计算组织学”地图。为了(A)告知发育阶段的数学模型和(B)大大增加从这些研究中获得的化学信息，分离的细胞将对数万个细胞进行另一种测量-高通量单细胞质谱仪(MS)。单细胞MS提供了每个细胞中数百种成分的详细信息，有效地绘制了每个细胞的多肽组和代谢组。MS数据包括关于这些细胞代谢状态的唯一信息，并允许我们定义已知和未知的细胞类型。计算模型将用于将SRSM数据与MS衍生的化学含量相关联，并提供检查脑组织动态变化和异质性的策略。这些技术将使用齿状回进行验证。工作的重点将集中在齿状回的海马神经元和胶质细胞及其参与记忆形成的过程，以及与星形胶质细胞形态变化相关的问题。通过对齿状回补丁细胞进行膜片钳生理测量和基于MS的详细代谢组学分析，SRSM和单细胞MS技术平台将通过研究这一包含多种细胞类型、不同形态和化学特征的复杂脑区来验证。这些技术将提供关于这个定义的大脑区域内化学成分和动态的无与伦比的细节，回答与细胞异质性有关的长期棘手的问题，并将这些信息与 组织和功能过程，如长时程增强。

 DESCRIPTION (provided by applicant): The BRAIN Initiative seeks to understand the spatial, temporal and chemical nature of the brain. RFA-MH-15- 225 calls for the development of new tools and technologies with a number of goals, including methods to obtain cell type and chemical information from individual cells and their connections. While many imaging approaches exist that use specific probes to image subsets of cells and their interconnections, this project will create a chemical information-rich approach that advances the emerging technique of stimulated Raman scattering microscopy (SRSM). SRSM provides vibrational spectral data from every location of a living brain slice so that dynamic chemical changes can be followed. The Raman spectra contain tremendous chemical information but the data is coded in complex overlapping molecular vibrational bands. With appropriate training sets-derived from the Raman data and comparing it to the chemical contents of individual cells-a series of mathematical models will be developed that create unlimited "Computational Histology" maps. In order to (a) inform the mathematical model in the development phase and (b) greatly augment the chemical information obtained from these studies, dissociated cells will be subjected to another measurement-high throughput single cell mass spectrometry (MS)-on tens of thousands of cells. Single cell MS provides detail on hundreds of components in each cell, effectively mapping each cells' peptidome and metabolome. The MS data includes unique information on the metabolic state of these cells and allows us to define known and unknown cell types. Computational models will be used to correlate the SRSM data to the MS-derived chemical content as well as deliver strategies to examine the dynamic changes and heterogeneity in brain tissue. These technologies will be validated using the dentate gyrus. The focus of the work will be on the hippocampal neurons and glia of the dentate gyrus and their involvement in memory formation, and issues related to astrocyte morphology changes. By performing patch clamp physiological measurements and detailed MS-based metabolomic profiling on the patched cells of the dentate gyrus, the SRSM and single cell MS technology platform will be validated by investigating this complex area of the brain containing many cell types, heterogeneous morphologies, and chemical characteristics. These technologies will provide unmatched detail on the chemical content and dynamics within this defined brain region, answer long intractable questions related to cellular heterogeneity, and relate this information to organization and functional processes such as long term potentiation.