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 细胞类型特异性标记的资源，正在彻底改变我们检测和监测年龄的能力， 疾病和神经连接的经验依赖性变化。基于荧光 定量突触分析的方法将很容易被科学界采用 并已广泛用于神经元培养，但尚未充分开发用于分析 大脑网络的连通性，由于对检测的分辨率和准确性的关注， 接触和试剂使用困难。在这里，我们将使用最近开发的工具， 突触标记开发工作流程，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在脑组织中进行基于测量。我们 然后将测试这些试剂的能力，以推动对特定输入可塑性的新见解 在学习过程中，使用一种特征良好的训练模式， 已经证实了丘脑皮质突触强度的变化。

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.