Collman Forrest, Buchanan JoAnn, Phend Kristen D, Micheva Kristina D, Weinberg Richard J, Smith Stephen J
Department of Molecular and Cellular Physiology, Stanford School of Medicine, Stanford University, Stanford California 94305, Allen Institute for Brain Science, Seattle, Washington 98103
Department of Molecular and Cellular Physiology, Stanford School of Medicine, Stanford University, Stanford California 94305.
J Neurosci. 2015 Apr 8;35(14):5792-807. doi: 10.1523/JNEUROSCI.4274-14.2015.
Synapses of the mammalian CNS are diverse in size, structure, molecular composition, and function. Synapses in their myriad variations are fundamental to neural circuit development, homeostasis, plasticity, and memory storage. Unfortunately, quantitative analysis and mapping of the brain's heterogeneous synapse populations has been limited by the lack of adequate single-synapse measurement methods. Electron microscopy (EM) is the definitive means to recognize and measure individual synaptic contacts, but EM has only limited abilities to measure the molecular composition of synapses. This report describes conjugate array tomography (AT), a volumetric imaging method that integrates immunofluorescence and EM imaging modalities in voxel-conjugate fashion. We illustrate the use of conjugate AT to advance the proteometric measurement of EM-validated single-synapse analysis in a study of mouse cortex.
哺乳动物中枢神经系统（CNS）的突触在大小、结构、分子组成和功能方面各不相同。突触的无数种变化形式对于神经回路的发育、稳态、可塑性和记忆存储至关重要。不幸的是，由于缺乏足够的单突触测量方法，对大脑中异质突触群体的定量分析和映射受到了限制。电子显微镜（EM）是识别和测量单个突触接触的权威方法，但EM在测量突触分子组成方面的能力有限。本报告描述了共轭阵列断层扫描（AT），这是一种以体素共轭方式整合免疫荧光和EM成像模式的容积成像方法。在一项对小鼠皮层的研究中，我们展示了使用共轭AT推进经EM验证的单突触分析的蛋白质组测量。
