Stefan Eimer1, Alexander Gottschalk2, Janet Richmond3, Michael Hengartner4, William R Schafer2, and Jean-Louis Bessereau1. Nicotinic acetylcholine receptors (nAChRs) are pentameric ligand-gated ion channels that undergo multiple assembly, modification and transport steps prior to insertion into the plasma membrane. However, little is known about the molecules that are specifically participating in these processes. In C. elegans a nAChR sensitive to the nematode-specific cholinergic agonist levamisole has been well characterised. Mutants in all of its four subunits confer resistance to muscle hyper-contraction and death upon exposure to levamisole.. Similarly,
unc-50 mutant animals exhibit the same resistance to paralysis and death upon exposure to levamisole. By antibody staining and electro-physiological recording of body-wall muscles we show that levamisole receptors are not detectable at cell surface in
unc-50 mutants. However, a second type of muscle nAChR, which is insensitive to levamisole, and the GABA receptor are normally transported to the synapse.
unc50 encodes an integral membrane protein conserved from yeast to man. In C. elegans,
unc-50 is ubiquitously expressed and localized to the Golgi system. We showed that UNC-50 is required in body wall muscles for the transport of the assembled levamisole receptor to neuromuscular junction. In the absence of UNC-50 the receptor is sorted within Golgi to the lysosomal system and rapidly degraded. Therefore, UNC-50 might be part of/constitute a Golgi resident check point during levamisole receptor transport. UNC-50 interacts with the GBF1 class of large Arf GEFs which are involved in retrograde transport within the Golgi system. Models how retrograde transport and sorting would required for levamisole receptor trafficking will be discussed.