Fluorescence-based methods for microconnectivity analysis in neocortex

基于荧光的新皮质微连接分析方法

基本信息

批准号：
10413555
负责人：
ALISON L BARTH
金额：
$ 159.7万
依托单位：
CARNEGIE-MELLON UNIVERSITY
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：
2022-05-01 至 2025-04-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10413555
关键词：
Address Adopted Age Anatomy Animals Antibodies Association Learning Axon Biological Brain Censuses Color Communities Data Data Set Dendrites Dendritic Spines Detection Development Disease Electron Microscopy Electrophysiology (science)Evaluation Excitatory Postsynaptic Potentials Fluorescence Functional disorder Future Image Individual Investigation Knowledge Label Laboratories Learning Light Link Measurement Methods Microscopy Molecular Molecular Genetics Monitor Neocortex Neurons Neurosciences Pattern Preparation Reagent Resolution Scientific Inquiry Sensory Specific qualifier value Specimen Speed Structure Synapses Synaptic plasticity Techniques Testing Thalamic structure Time Tissues Training Validation barrel cortex base brain tissue cell type confocal imaging density experience genetic resource hippocampal pyramidal neuron in vivo information processing insight instrumentation microscopic imaging mouse model nanometer resolution neural circuit neural network postsynaptic postsynaptic neurons presynaptic reconstruction relating to nervous system sensorimotor system tool tool development transmission process two photon microscopy

项目摘要

ABSTRACT A comprehensive census of neural cell types in the brain, together with molecular-genetic resources for cell-type specific labeling, is revolutionizing our ability to detect and monitor age, disease, and experience-dependent changes in neural connectivity. Fluorescence-based methods for quantitative synapse analysis would be easily adopted by the scientific community and have been widely used in neuronal cultures, but have not been well-developed for analysis of connectivity in brain networks, due to concerns about resolution and accuracy of detected contacts and difficulty of reagent use. Here we will use recently-developed tools for fluorescent synaptic labeling to develop a workflow to evaluate and mitigate concerns about fluorescence- based measurements in brain tissue using state-of-the-art Expansion Microscopy methods. We will then test the ability of these reagents to drive new insights about input-specific plasticity during learning, using a well-characterized training paradigm where electrophysiological changes in thalamocortical synaptic strength have been demonstrated.

抽象的大脑中神经细胞类型的全面人口普查以及分子遗传学用于细胞类型的特定标签的资源，正在革新我们检测和监视年龄的能力，疾病和经验依赖神经连通性的变化。基于荧光科学界很容易采用定量突触分析的方法并已被广泛用于神经元文化，但尚未得到充分发达的分析由于担心分辨率和检测到的准确性，大脑网络中的连通性接触和使用试剂的难度。在这里，我们将使用最近开发的工具进行荧光突触标记以开发工作流程，以评估和减轻对荧光的担忧使用最先进的扩展显微镜方法中的基于脑组织的测量。我们然后，将测试这些试剂的能力，以推动有关特定于输入的可塑性的新见解在学习过程中，使用电流生理的特征训练范式已经证明了丘脑皮质突触强度的变化。