Abstract
Analgesic neurosteroids such as 5alpha-pregnan-3alpha-ol-20-one (5alpha3alpha) are potent selective endogenous modulators of the GABA(A) receptor (GABA(A)R) while certain synthetic derivatives (i.e. minaxolone) additionally enhance the function of recombinant glycine receptors (GlyR). Inhibitory transmission within the superficial dorsal horn has been implicated in mediating the analgesic actions of neurosteroids. However, the relative contribution played by synaptic and extrasynaptic receptors is unknown. In this study, we have compared the actions of 5alpha3alpha and minaxolone upon inhibitory transmission mediated by both GABA(A) and strychnine-sensitive GlyRs in lamina II neurones of juvenile (P15-21) rats. At the near physiological temperature of 35 degrees C and at a holding potential of -60 mV we recorded three kinetically distinct populations of miniature IPSCs (mIPSCs): GlyR-mediated, GABA(A)R-mediated and mixed GABA(A)R-GlyR mIPSCs, arising from the corelease of both inhibitory neurotransmitters. In addition, sequential application of strychnine and bicuculline revealed a small (5.2 +/- 1.0 pA) GlyR- but not a GABA(A)R-mediated tonic conductance. 5alpha3alpha (1-10 microm) prolonged GABA(A)R and mixed mIPSCs in a concentration-dependent manner but was without effect upon GlyR mIPSCs. In contrast, minaxolone (1-10 microm) prolonged the decay of GlyR mIPSCs and, additionally, was approximately 10-fold more potent than 5alpha3alpha upon GABA(A)R mIPSCs. However, 5alpha3alpha and minaxolone (1 microm) evoked a similar bicuculline-sensitive inhibitory conductance, indicating that the extrasynaptic GABA(A)Rs do not discriminate between these two steroids. Furthermore, approximately 92% of the effect of 1 microm 5alpha3alpha upon GABAergic inhibition could be accounted for by its action upon the extrasynaptic conductance. These findings are relevant to modulation of inhibitory circuits within spinally mediated pain pathways and suggest that extrasynaptic GABA(A)Rs may represent a relevant molecular target for the analgesic actions of neurosteroids.