Association of Myosin Va and Schwann cells-derived RNA in mammal myelinated axons, analyzed by immunocytochemistry and confocal FRET microscopy

Evidence from multiple sources supports the hypothesis that Schwann cells in the peripheral nervous system transfer messenger RNA and ribosomes to the axons they ensheath. Several technical and methodological difficulties exist for investigators to unravel this process in myelinated axons – a comple...

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Veröffentlicht in:	Methods (San Diego, Calif.) Calif.), 2014-03, Vol.66 (2), p.153-161
Hauptverfasser:	Canclini, Lucía, Wallrabe, Horst, Di Paolo, Andrés, Kun, Alejandra, Calliari, Aldo, Sotelo-Silveira, José Roberto, Sotelo, José Roberto
Format:	Artikel
Sprache:	eng
Schlagworte:	Animals Axon Axons - metabolism Fluorescence Resonance Energy Transfer Immunohistochemistry Microscopy, Confocal Myosin Heavy Chains - metabolism Myosin Type V - metabolism Myosin Va Peripheral Nerves - cytology Peripheral Nerves - metabolism Quantitative FRET microscopy Ranvier's Nodes - metabolism Rats RNA cell-to-cell transfer RNA Transport RNA, Messenger - metabolism Schwann cell Schwann Cells - metabolism
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Zusammenfassung:	Evidence from multiple sources supports the hypothesis that Schwann cells in the peripheral nervous system transfer messenger RNA and ribosomes to the axons they ensheath. Several technical and methodological difficulties exist for investigators to unravel this process in myelinated axons – a complex two-cell unit. We present an experimental design to demonstrate that newly synthesized RNA is transferred from Schwann cells to axons in association with Myosin Va. The use of quantitative confocal FRET microscopy to track newly-synthesized RNA and determine the molecular association with Myosin Va, is described in detail.
ISSN:	1046-2023 1095-9130
DOI:	10.1016/j.ymeth.2013.06.007