In Vivo Synaptic Imaging in Neocortex

新皮质体内突触成像

基本信息

批准号：
10471169
负责人：
ALISON L BARTH
金额：
$ 22.55万
依托单位：
CARNEGIE-MELLON UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-09-01 至 2024-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10471169
关键词：
Anatomy Apical Axon Brain Cell Nucleus Cells Chemosensitization Color Cortical Column Coupled Data Dendrites Dendritic Spines Detection Electrophysiology (science)Excitatory Synapse Fluorescence Foundations Glutamates Histologic Image Image Analysis Immunohistochemistry Label Learning Learning Disorders Link Medial Memory Disorders Methodology Methods Microscopy Molecular Molecular Genetics Monitor Mus Neocortex Pathway interactions Preparation Property Proteins Resolution Rewards Sampling Sensory Site Somatosensory Cortex Synapses Synaptic plasticity Testing Thalamic Nuclei Thalamic structure Tissues Training Validation Vertebral column Vibrissae analysis pipeline barrel cortex base cell type experience hippocampal pyramidal neuron in vivo in vivo fluorescence imaging in vivo imaging in vivo monitoring information processing insight nanoscale neocortical neural circuit patch clamp photon-counting detector postsynaptic postsynaptic neurons presynaptic relating to nervous system scaffold sensorimotor system serial imaging somatosensory two-photon

项目摘要

ABSTRACT Synapse addition and loss have been linked to learning throughout the brain. This has been well-documented in the neocortex using anatomical analysis of axonal boutons or dendritic spines in both fixed tissue and in vivo, using longitudinal imaging. However, it has been difficult to extrapolate the consequences of these synaptic changes without understanding how they lie within a defined network of cell-type specific contacts, requiring identification of the pre- and postsynaptic partners of the synapse. Here we will use in vivo imaging and multicolor fluorescence labeling of molecularly-defined pre- and postsynaptic neurons to monitor input- specific synapse addition and loss during sensory learning in a whisker-dependent task in mice. Synaptic contacts will be validated using post-hoc expansion microscopy and immunohistochemistry for nanoscale resolution. Analysis will be focused on learning-dependent reorganization of thalamic inputs from a higher-order thalamic nucleus, the posterior medial nucleus (POm) onto the dendrites of layer 5 (L5) pyramidal neurons that have been specifically implicated in experience-dependent plasticity. Quantitative, multicolor, in vivo imaging across different stages of learning will provide insight into how cortical circuits encode and are changed by salient sensory information.

抽象的突触的增加和丧失与整个大脑的学习有关。这已经是使用轴突或树突的解剖分析在新皮质中有详细记录使用纵向成像在固定组织和体内的脊柱中。然而，这已经很难了在不了解这些突触变化的原理的情况下推断它们的后果在细胞类型特定接触的定义网络内，需要识别预和突触的突触后伙伴。这里我们将使用体内成像和多色分子定义的突触前和突触后神经元的荧光标记以监测输入小鼠胡须依赖性任务中感觉学习过程中特定突触的添加和丢失。突触接触将使用事后扩展显微镜进行验证纳米级分辨率的免疫组织化学。分析将侧重于学习依赖来自高阶丘脑核（后内侧核）的丘脑输入的重组核（POm）到第5层（L5）锥体神经元的树突上，这些神经元已被专门涉及依赖于经验的可塑性。定量、多色、体内成像不同阶段的学习将深入了解皮质回路如何编码和改变通过显着的感官信息。