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损失与整个大脑的学习有关。这是使用轴突牙骨或树突状的解剖学分析在新皮层中有充分的文献记录使用纵向成像，固定组织和体内的棘突。但是，很难推断这些突触变化的后果而不了解它们如何说谎在定义的细胞类型特定联系网络中，需要识别前和突触的突触后伴侣。在这里，我们将使用体内成像和多色分子定义的前和突触后神经元的荧光标记，以监测输入在小鼠的晶须依赖性任务中，在感觉学习过程中的特定突触添加和损失。突触接触将使用事后膨胀显微镜进行验证，并纳米级分辨率的免疫组织化学。分析将集中于学习依赖性来自高阶丘脑核的丘脑输入的重组，后内侧核（POM）到第5层（L5）锥体神经元的树突上与经验依赖性可塑性有关。定量，多色，体内成像不同的学习阶段将提供有关皮质电路如何编码和更改的洞察力通过显着的感官信息。