Axoplasm isolation from peripheral nerve |
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| Authors: | Ida Rishal Izhak Michaelevski Meir Rozenbaum Vera Shinder Katalin F Medzihradszky Alma L Burlingame Mike Fainzilber |
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| Institution: | 1. Department of Biological Chemistry, Weizmann Institute of Science, 76100 Rehovot, Israel;2. Electron Microscopy Unit, Department of Chemical Research Support, Weizmann Institute of Science, 76100 Rehovot, Israel;3. Mass Spectrometry Facility, Department of Pharmaceutical Chemistry, University of California San Francisco, San Francisco, CA 94158‐2517 |
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| Abstract: | Localized changes in the composition of axonal cytoplasm (axoplasm) are critical for many biological processes, including axon guidance, responses to injury, neurite outgrowth, and axon‐glia interactions. Biochemical and molecular studies of these mechanisms have been heavily focused on in vitro systems because of the difficulty of obtaining subcellular extracts from mammalian tissues in vivo. As in vitro systems might not replicate the in vivo situation, reliable methods of axoplasm extraction from whole nerve would be helpful for mechanistic studies on axons. Here we develop and evaluate a new procedure for preparation of axoplasm from rat peripheral nerve, based on incubation of separated short segements of nerve fascicles in hypotonic medium to separate myelin and lyse nonaxonal structures, followed by extraction of the remaining axon‐enriched material. We show that this new procedure reduces serum and glial cell contamination and facilitates proteomic analyses of axonal contents. © 2009 Wiley Periodicals, Inc. Develop Neurobiol, 2010 |
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| Keywords: | axoplasm axon proteomics |
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