A neomorphic syntaxin mutation blocks volatile-anesthetic action in Caenorhabditis elegans.
The molecular mechanisms underlying general anesthesia are unknown. For volatile general anesthetics (VAs), indirect evidence for both lipid and protein targets has been found. However, no in vivo data have implicated clearly any particular lipid or protein in the control of sensitivity to clinical concentrations of VAs. Genetics provides one approach toward identifying these mechanisms, but genes strongly regulating sensitivity to clinical concentrations of VAs have not been identified. By screening existing mutants of the nematode Caenorhabditis elegans, we found that a mutation in the neuronal syntaxin gene dominantly conferred resistance to the VAs isoflurane and halothane. By contrast, other mutations in syntaxin and in the syntaxin-binding proteins synaptobrevin and SNAP-25 produced VA hypersensitivity. The syntaxin allelic variation was striking, particularly for isoflurane, where a 33-fold range of sensitivities was seen. Both the resistant and hypersensitive mutations decrease synaptic transmission; thus, the indirect effect of reducing neurotransmission does not explain the VA resistance. As assessed by pharmacological criteria, halothane and isoflurane themselves reduced cholinergic transmission, and the presynaptic anesthetic effect was blocked by the resistant syntaxin mutation. A single gene mutation conferring high-level resistance to VAs is inconsistent with nonspecific membrane-perturbation theories of anesthesia. The genetic and pharmacological data suggest that the resistant syntaxin mutant directly blocks VA binding to or efficacy against presynaptic targets that mediate anesthetic behavioral effects. Syntaxin and syntaxin-binding proteins are candidate anesthetic targets.
Department of Anesthesiology, Division of Biology and Biomedical Sciences, Washington University School of Medicine, 660 South Euclid Avenue, St. Louis, MO 63110, USA.
B
B
van Swinderen
O
O
Saifee
L
L
Shebester
R
R
Roberson
M L
ML
Nonet
C M
CM
Crowder
D000448:Aldicarb / Q000494:pharmacology; D000595:Amino Acid Sequence; D018685:Anesthetics, Inhalation / Q000494:pharmacology*; D000818:Animals; D001483:Base Sequence; D002107:Caenorhabditis / Q000187:drug effects / Q000235:genetics*; D003433:Crosses, Genetic; D012734:Disorders of Sex Development; D005799:Genes, Dominant; D017238:Genes, Helminth; D006221:Halothane / Q000494:pharmacology; D007530:Isoflurane / Q000494:pharmacology; D008565:Membrane Proteins / Q000737:chemistry / Q000235:genetics* / Q000502:physiology; D008969:Molecular Sequence Data; D009154:Mutation; D009419:Nerve Tissue Proteins / Q000235:genetics / Q000502:physiology; D010641:Phenotype; D050765:Qa-SNARE Proteins; D012044:Regression Analysis; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D009435:Synaptic Transmission / Q000502:physiology; D016708:Synaptophysin / Q000235:genetics / Q000502:physiology; D050825:Synaptosomal-Associated Protein 25
D018685:Anesthetics, Inhalation; D008565:Membrane Proteins; D009419:Nerve Tissue Proteins; D050765:Qa-SNARE Proteins; D016708:Synaptophysin; D050825:Synaptosomal-Associated Protein 25; D000448:Aldicarb; D007530:Isoflurane; D006221:Halothane