Syntaxin1a Neomorphic Mutations Promote Rapid Recovery From Isoflurane Anesthesia In Drosophila Melanogaster

Background: Mutations in the presynaptic protein syntaxin1A modulate general anesthetic effects in vitro and in vivo. Coexpression of a truncated syntaxin1A protein confers resistance to volatile and intravenous anesthetics, suggesting a target mechanism distinct from postsynaptic inhibitory receptor processes. Hypothesizing that recovery from anesthesia may involve a presynaptic component, the authors tested whether syntaxin1A mutations facilitated recovery from isoflurane anesthesia in Drosophila melanogaster.Methods: A truncated syntaxin1A construct was expressed in Drosophila neurons. The authors compared effects on isoflurane induction versus recovery in syntaxin1A mutant animals by probing behavioral responses to mechanical stimuli. The authors also measured synaptic responses from the larval neuromuscular junction using sharp intracellular recordings, and performed Western blots to determine whether the truncated syntaxin1A is associated with presynaptic core complexes.Results: Drosophila expressing a truncated syntaxin1A (syx(227), n = 40) were resistant to isoflurane induction for a behavioral responsiveness endpoint (ED50 0.30 0.01% isoflurane, P < 0.001) compared with control (0.240 +/- 0.002% isoflurane, n = 40). Recovery from isoflurane anesthesia was also faster, with syx(227)-expressing flies showing greater levels of responsiveness earlier in recovery (reaction proportion 0.66 +/- 0.48, P < 0.001, n = 68) than controls (0.22 +/- 0.42, n = 68 and 0.33 +/- 0.48, n = 66). Measuring excitatory junction potentials of larvae coexpressing the truncated syntaxin1A protein showed a greater recovery of synaptic function, compared with controls (17.39 +/- 3.19 mV and 10.29 +/- 4.88 mV, P = 0.014, n = 8 for both). The resistance-promoting truncated syntaxin1A was not associated with presynaptic core complexes, in the presence or absence of isoflurane anesthesia.Conclusions: The same neomorphic syntaxin1A mutation that confers isoflurane resistance in cell culture and nematodes also produces isoflurane resistance in Drosophila. Resistance in Drosophila is, however, most evident at the level of recovery from anesthesia, suggesting that the syntaxin1A target affects anesthesia maintenance and recovery processes rather than induction. The absence of truncated syntaxin1A from the presynaptic complex suggests that the resistance-promoting effect of this molecule occurs before core complex formation.