UNC-41/stonin functions with AP2 to recycle synaptic vesicles in Caenorhabditis elegans

PLoS One. 2012;7(7):e40095. doi: 10.1371/journal.pone.0040095. Epub 2012 Jul 10.

Abstract

The recycling of synaptic vesicles requires the recovery of vesicle proteins and membrane. Members of the stonin protein family (Drosophila Stoned B, mammalian stonin 2) have been shown to link the synaptic vesicle protein synaptotagmin to the endocytic machinery. Here we characterize the unc-41 gene, which encodes the stonin ortholog in the nematode Caenorhabditis elegans. Transgenic expression of Drosophila stonedB rescues unc-41 mutant phenotypes, demonstrating that UNC-41 is a bona fide member of the stonin family. In unc-41 mutants, synaptotagmin is present in axons, but is mislocalized and diffuse. In contrast, UNC-41 is localized normally in synaptotagmin mutants, demonstrating a unidirectional relationship for localization. The phenotype of snt-1 unc-41 double mutants is stronger than snt-1 mutants, suggesting that UNC-41 may have additional, synaptotagmin-independent functions. We also show that unc-41 mutants have defects in synaptic vesicle membrane endocytosis, including a ∼50% reduction of vesicles in both acetylcholine and GABA motor neurons. These endocytic defects are similar to those observed in apm-2 mutants, which lack the µ2 subunit of the AP2 adaptor complex. However, no further reduction in synaptic vesicles was observed in unc-41 apm-2 double mutants, suggesting that UNC-41 acts in the same endocytic pathway as µ2 adaptin.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adaptor Protein Complex 2 / metabolism*
  • Adaptor Proteins, Vesicular Transport / genetics
  • Adaptor Proteins, Vesicular Transport / metabolism*
  • Animals
  • Animals, Genetically Modified
  • Caenorhabditis elegans / genetics
  • Caenorhabditis elegans / metabolism
  • Caenorhabditis elegans / ultrastructure
  • Caenorhabditis elegans Proteins / genetics
  • Caenorhabditis elegans Proteins / metabolism*
  • Carrier Proteins / metabolism
  • Cloning, Molecular
  • Drosophila Proteins / metabolism
  • Endocytosis*
  • Gene Expression Regulation
  • Genes, Helminth / genetics
  • Genome / genetics
  • Mutation / genetics
  • Nerve Tissue Proteins / metabolism
  • Nervous System / metabolism
  • Phenotype
  • Protein Transport
  • Synaptic Vesicles / metabolism*
  • Synaptic Vesicles / ultrastructure
  • Synaptotagmins / metabolism
  • Vesicular Transport Proteins

Substances

  • Adaptor Protein Complex 2
  • Adaptor Proteins, Vesicular Transport
  • Caenorhabditis elegans Proteins
  • Carrier Proteins
  • Drosophila Proteins
  • Nerve Tissue Proteins
  • Unc-41 protein, C elegans
  • Vesicular Transport Proteins
  • stnB protein, Drosophila
  • Synaptotagmins