Fluorescence-based methods for microconnectivity analysis in neocortex

摘要 对大脑中神经细胞类型的全面普查，以及分子遗传学 用于细胞类型特定标记的资源正在彻底改变我们检测和监测年龄的能力， 疾病，以及神经连接性的经验依赖性变化。基于荧光的 定量分析突触的方法将很容易被科学界采用 已经被广泛应用于神经元培养，但还没有被很好地开发用于分析 由于对检测到的分辨率和准确性的担忧，大脑网络中的连通性 接触和试剂使用困难。在这里，我们将使用最近开发的荧光工具 突触标记以开发一种工作流程来评估和缓解对荧光的担忧- 使用最先进的扩展显微镜方法对脑组织进行测量。我们 然后将测试这些试剂推动对输入特定可塑性的新见解的能力 在学习过程中，使用具有良好特征的训练范例，其中电生理学 丘脑皮质突触强度的变化已经被证明。

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.