Electron Tomographic Methods for Studying the Chemical Synapse

• Published in: Methods in Cell Biology Vol. 79; pp. 241 - 257
• Main Authors: Arthur, Christopher P., Serrell, David B., Pagratis, Maria, Potter, David L., Finch, Dudley S., Stowell, Michael H.B.
• Format: Book Chapter Journal Article
• Language: English
• Published: United States Elsevier Science & Technology 2007
• Subjects: Animals; Humans; Microscopy, Electron - methods; Models, Biological; Nerve Net - ultrastructure; Neurons - ultrastructure; Synapses - ultrastructure
• ISBN: 0123706475; 9780123706478
• ISSN: 0091-679X
• DOI: 10.1016/S0091-679X(06)79010-0

This chapter focuses on the use of electron tomography (ET) to study the chemical synapse. It discusses the different types of chemical synapses and their properties as well as the biological questions that are addressable using ET. It also discusses the methods for culturing neurons on a variety of substrates suitable for ET and the development of novel tools that combine electrophysiological methods with ET methods for studying active synapses. The ultimate goal of such studies is to identify and ascertain the organization of membranes and supramolecular complexes in a resting synapse or in a synapse after treatment with well-defined stimulation protocols. Many of the relevant molecular complexes are of sufficient mass and their appearance causes clear contrast in conventional electron microscopy (EM). These include important assemblies such as the SNARE complex, NSF, and NMDA-PSD-95, and the bassoon and piccolo complexes that order synaptic vesicles, polysomes, Ca2+-release channels, microtubules, and associated transport vesicles. The study of several systems, such as the neuromuscular junction and ribbon synapses following stimulation, has been achieved and important insights have been gained; such studies cannot currently be performed with primary cultured hippocampus neurons.