Insights into the Molecular Organization of the Neuron by Cryo-Electron Tomography
JOURNAL OF ELECTRON MICROSCOPY(2011)
Abstract
Despite great progress in the identification and characterization of the key molecular players in neuronal function, remarkably little is known about their supramolecular organization. Cryo-electron tomography (cryo-ET), providing three-dimensional views of the molecular components of the cell in their native, fully hydrated environment, is uniquely positioned to elucidate the native architecture of the molecular machinery of the neuron. In our laboratory, we employ cryo-ET to study neuronal morphology in a variety of experimental systems and develop methods to extract quantitative and functional information from tomographic data. This approach has allowed us to shed light onto the intricate organization of the molecules of the synaptic cleft and the presynaptic cytomatrix, providing evidence for their functional roles. Also, cryo-ET of cultured neurons is beginning to open new perspectives on neuronal ultrastructure and the architecture of synaptic complexes in situ. Here, we will review these findings and discuss future directions towards the elucidation of the molecular landscape of the neuron.
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Key words
cryo-electron tomography,neuron,synapse,presynaptic cytomatrix,automated segmentation,correlative microscopy
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