Wang Gordon X, Smith Stephen J, Mourrain Philippe
Department of Psychiatry and Behavioral Sciences, Stanford University School of Medicine, Stanford, United States.
Center for Sleep Sciences and Medicine, Stanford University School of Medicine, Stanford, United States.
Elife. 2016 Oct 22;5:e20560. doi: 10.7554/eLife.20560.
The distribution of proteins within sub-synaptic compartments is an essential aspect of their neurological function. Current methodologies, such as electron microscopy (EM) and super-resolution imaging techniques, can provide the precise localization of proteins, but are often limited to a small number of one-time observations with narrow spatial and molecular coverage. The diversity of synaptic proteins and synapse types demands synapse analysis on a scale that is prohibitive with current methods. Here, we demonstrate SubSynMAP, a fast, multiplexed sub-synaptic protein analysis method using wide-field data from deconvolution array tomography (ATD). SubSynMAP generates probability distributions for that reveal the functional range of proteins within the averaged synapse of a particular class. This enables the differentiation of closely juxtaposed proteins. Using this method, we analyzed 15 synaptic proteins in normal and Fragile X mental retardation syndrome (FXS) model mouse cortex, and revealed disease-specific modifications of sub-synaptic protein distributions across synapse classes and cortical layers.
蛋白质在突触亚区室中的分布是其神经功能的一个重要方面。当前的方法，如电子显微镜（EM）和超分辨率成像技术，可以提供蛋白质的精确定位，但通常限于少量一次性观察，空间和分子覆盖范围狭窄。突触蛋白和突触类型的多样性要求在当前方法难以企及的规模上进行突触分析。在这里，我们展示了SubSynMAP，一种使用去卷积阵列断层扫描（ATD）的宽场数据进行快速、多重亚突触蛋白分析的方法。SubSynMAP生成概率分布，揭示特定类别平均突触内蛋白质的功能范围。这使得能够区分紧密相邻的蛋白质。使用这种方法，我们分析了正常和脆性X智力障碍综合征（FXS）模型小鼠皮质中的15种突触蛋白，并揭示了跨突触类别和皮质层的亚突触蛋白分布的疾病特异性修饰。
