Functional reconstitution of Drosophila melanogaster NMJ glutamate receptors.
The Drosophila larval neuromuscular junction (NMJ), at which glutamate acts as the excitatory neurotransmitter, is a widely used model for genetic analysis of synapse function and development. Despite decades of study, the inability to reconstitute NMJ glutamate receptor function using heterologous expression systems has complicated the analysis of receptor function, such that it is difficult to resolve the molecular basis for compound phenotypes observed in mutant flies. We find that Drosophila Neto functions as an essential component required for the function of NMJ glutamate receptors, permitting analysis of glutamate receptor responses in Xenopus oocytes. In combination with a crystallographic analysis of the GluRIIB ligand binding domain, we use this system to characterize the subunit dependence of assembly, channel block, and ligand selectivity for Drosophila NMJ glutamate receptors.
Program in Cellular Regulation and Metabolism and.
Tae Hee
TH
Han
Laboratory of Cellular and Molecular Neurophysiology, The Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892.
Poorva
P
Dharkar
Laboratory of Cellular and Molecular Neurophysiology, The Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892 mayerm@mail.nih.gov mihaela.serpe@nih.gov.
Mark L
ML
Mayer
Program in Cellular Regulation and Metabolism and mayerm@mail.nih.gov mihaela.serpe@nih.gov.
Mihaela
M
Serpe
D000595:Amino Acid Sequence; D000818:Animals; D002118:Calcium / Q000378:metabolism; D002462:Cell Membrane / Q000378:metabolism; D018360:Crystallography, X-Ray; D018076:DNA, Complementary / Q000378:metabolism; D029721:Drosophila Proteins / Q000235:genetics* / Q000502:physiology; D004331:Drosophila melanogaster / Q000235:genetics*; D007477:Ions; D008024:Ligands; D008565:Membrane Proteins / Q000235:genetics* / Q000502:physiology; D018613:Microscopy, Confocal; D008969:Molecular Sequence Data; D009154:Mutation; D009469:Neuromuscular Junction / Q000502:physiology*; D009865:Oocytes / Q000166:cytology / Q000378:metabolism; D010539:Permeability; D010641:Phenotype; D017434:Protein Structure, Tertiary; D017470:Receptors, Glutamate / Q000378:metabolism*; D017386:Sequence Homology, Amino Acid; D013569:Synapses / Q000502:physiology; D014982:Xenopus laevis
D018076:DNA, Complementary; D029721:Drosophila Proteins; D007477:Ions; D008024:Ligands; D008565:Membrane Proteins; C573279:Neto protein, Drosophila; D017470:Receptors, Glutamate; D002118:Calcium
Drosophila; NMJ; Neto; crystallography; glutamate receptors