Effects of nitrous oxide on glycinergic transmission in rat spinal neurons

We investigated the effects of nitrous oxide (N₂O) on glycinergic inhibitory whole-cell and synaptic responses using a "synapse bouton preparation," dissociated mechanically from rat spinal sacral dorsal commissural nucleus (SDCN) neurons. This technique can evaluate pure single- or multi-synaptic responses from native functional nerve endings and enable us to accurately quantify how N₂O influences pre- and postsynaptic transmission. We found that 70 % N₂O enhanced exogenous glycine-induced whole-cell currents (I_Gly) at glycine concentrations lower than 3 × 10^-5 M, but did not affect I_Gly at glycine concentrations higher than 10^-4 M. N₂O did not affect the amplitude and 1/e decay-time of both spontaneous and miniature glycinergic inhibitory postsynaptic currents recorded in the absence and presence of tetrodotoxin (sIPSCs and mIPSCs, respectively). The decrease in frequency induced by N₂O was observed in sIPSCs but not in mIPSCs, which was recorded in the presence of both tetrodotoxin and Cd²⁺, which block voltage-gated Na⁺ and Ca²⁺ channels, respectively. N₂O also decreased the amplitude and increased the failure rate and paired-pulse ratio of action potential-evoked glycinergic inhibitory postsynaptic currents. N₂O slightly decreased the Ba²⁺ currents mediated by voltage-gated Ca²⁺ channels in SDCN neurons. We found that N₂O suppresses glycinergic responses at synaptic levels with presynaptic effect having much more predominant role. The difference between glycinergic whole-cell and synaptic responses suggests that extrasynaptic responses seriously modulate whole-cell currents. Our results strongly suggest that these responses may thus in part explain analgesic effects of N₂O via marked glutamatergic inhibition by glycinergic responses in the spinal cord.