Acetylcholine Receptors Direct Rapsyn Clusters to the Neuromuscular Synapse in Zebrafish

Clustering of nicotinic muscle acetylcholine receptors (AChRs) requires association with intracellular rapsyn, a protein with an intrinsic ability to self-cluster. Previous studies on sofa potato (sop), an AChR null line of zebrafish, have suggested that AChRs may play an active role in subsynaptic localization of rapsyn clusters. To test this proposal directly, we identified and cloned the gene responsible for the sop phenotype and then attempted to rescue subsynaptic localization of the receptor-rapsyn complex in mutant fish. sop contains a leucine to proline mutation at position 28, near the N terminus of the zebrafish AChR δ subunit. Transient expression of mutant δ subunit in sop fish was unable to restore surface expression of muscle AChRs. In contrast, expression of wild-type δ subunit restored the ability of muscle to assemble surface receptors along with the ability of fish to swim. Most importantly, the ability of rapsyn clusters to localize effectively to subsynaptic sites also was rescued in large part. Our results point to direct involvement of the AChR molecule in restricting receptor-rapsyn clusters to the synapse.

• Publication year

  2004

• Venue

  Journal of Neuroscience

• Publication date

  2004-06-16

• Fields of study

  Biology, Medicine

• Identifiers
  DOI 10.1523/JNEUROSCI.0851-04.2004PMID 15201319PMCID PMC6729331
• External record

  Open on Semantic Scholar

• Source metadata

  Semantic Scholar, PubMed

• No claims are published for this paper.

• No concepts are published for this paper.

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  2002influential reference
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  2001cited by this paper
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  1999cited by this paper
• Congenital myasthenic syndrome caused by novel loss-of-function mutations in the human AChR epsilon subunit gene.

  1998cited by this paper
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  1998cited by this paper
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  1998cited by this paper
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  1998influential reference
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  1998influential reference
• Deficient development and maintenance of postsynaptic specializations in mutant mice lacking an 'adult' acetylcholine receptor subunit.

  1997influential reference
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  1996influential reference
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  1996cited by this paper
• End-plate acetylcholine receptor deficiency due to nonsense mutations in the epsilon subunit.

  1996cited by this paper
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  1996cited by this paper
• End‐plate acetylcholine receptor deficiency due to nonsense mutations in the ε subunit

  1996cited by this paper
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  1991cited by this paper
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  1990cited by this paper
• Pathfinding and synapse formation in a zebrafish mutant lacking functional acetylcholine receptors.

  1990cited by this paper
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  1990cited by this paper
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  1985cited by this paper

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