3-Dimensional Reconstruction of Parallel Fiber-Purkinje Cell Synapses Using High-Voltage Electron Microscopy

摘要: Synapses are contact points where one neuron communicates with another. The morphological change of synapses under various physiological or pathological conditions has long been hypothesized to modify their functional properties. 3-dimensional (3-D) reconstruction serial ultrathin sections contributed the understanding ultrastructural dimensions and compositions synapses. 3-D procedures, however, require a great amount expertise as well include prohibitively timeconsuming processes. Here, we introduce efficient technique using high-voltage electron microscopy (HVEM). Primarily, established an optimal section thickness staining condition observe synaptic structures in detail HVEM. result showed that profiles were preserved at 250 nm without overlapping ultrastructures. An increase reaction time en bloc was most enhance contrast than extension postembedding addition uranyl acetate during dehydration. Then, parallel fiber-Purkinje cell rat cerebellum carried out HVEM images software. aligned contours outlined on each section. finally extracted from files by grouping all contours. reconstructed synapse model clearly demonstrated configuration pre postsynaptic components. These results suggest is much suitable for massive quantitative studies connectivity conventional TEM approach numerous sections.