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  Assembly and clustering of acetylcholine receptors containing GFP-tagged epsilon or gamma subunits: Selective targeting to the neuromuscular junction in vivo

Gensler, S., Sander, A., Korngreen, A., Traina, G., Giese, G., & Witzemann, V. (2001). Assembly and clustering of acetylcholine receptors containing GFP-tagged epsilon or gamma subunits: Selective targeting to the neuromuscular junction in vivo. European Journal of Biochemistry, 268(8), 2209-2217. doi:10.1046/j.1432-1327.2001.02093.x.

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Item Permalink: http://hdl.handle.net/11858/00-001M-0000-0028-EB8B-3 Version Permalink: http://hdl.handle.net/11858/00-001M-0000-0028-EB8C-1
Genre: Journal Article
Alternative Title : Assembly and clustering of acetylcholine receptors containing GFP-tagged epsilon or gamma subunits: Selective targeting to the neuromuscular junction in vivo

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EurJBiochem_268_2001_ 2209.pdf (Any fulltext), 690KB
 
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 Creators:
Gensler, Sven1, Author              
Sander, Andreas1, Author              
Korngreen, Alon1, Author              
Traina, Giovanna, Author
Giese, Günter2, Author              
Witzemann, Veit1, 3, Author              
Affiliations:
1Department of Cell Physiology, Max Planck Institute for Medical Research, Max Planck Society, ou_1497701              
2Department of Biomedical Optics, Max Planck Institute for Medical Research, Max Planck Society, ou_1497699              
3Department of Molecular Neurobiology, Max Planck Institute for Medical Research, Max Planck Society, ou_1497704              

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Free keywords: acetylcholine receptor; GFP; assembly; targeting; neuromuscular junction
 Abstract: Acetylcholine receptor (AChR) gamma and epsilon subunits were tagged by green fluorescent protein (GFP) to analyse assembly and targeting in live muscle fibers at the neuromuscular junction. N- or C-terminal fusion polypeptides showed no fluorescence upon transfection of HEK cells. When GFP was inserted into the cytoplasmic loop connecting putative transmembrane regions M3 and M4, the gamma/GFP and epsilon/GFP subunits were fluorescent and formed together with the alpha, beta, and delta subunits GFP-tagged AChR complexes that were integrated into the plasma membrane. As the AChR were also clustered by rapsyn, the results indicate that the cytoplasmatic domains of the gamma and epsilon subunits may not be required for assembly and rapsyn-dependent clustering. The gamma/GFP and epsilon/GFP subunit-containing receptors were expressed in X. laevis oocytes and have affinities for acetylcholine similar to that of the wild-type receptors. Direct gene transfer into single muscle fibers reveals that gamma/GFP or epsilon/GFP polypeptides are expressed at the site of injection and are transported within the endoplasmatic reticulum. When reaching subsynaptic regions, both gamma/GFP or epsilon/GFP subunits compete with endogenous epsilon subunits to assemble GFP-tagged receptors, which are selectively targeted to the postsynaptic membrane.

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Language(s): eng - English
 Dates: 2001-12-202001-02-122001-12-202001-04-01
 Publication Status: Published in print
 Pages: 9
 Publishing info: -
 Table of Contents: -
 Rev. Method: Peer
 Degree: -

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Title: European Journal of Biochemistry
Source Genre: Journal
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Publ. Info: Berlin : Published by Springer-Verlag on behalf of the Federation of European Biochemical Societies
Pages: - Volume / Issue: 268 (8) Sequence Number: - Start / End Page: 2209 - 2217 Identifier: ISSN: 0014-2956
CoNE: https://pure.mpg.de/cone/journals/resource/111097776606040