strychnine and tetrahydrodeoxycorticosterone

The sympathetic tone is primarily defined by the level of activity of the sympathetic preganglionic neurons. We report a novel inhibitory influence on sympathetic activity, that of tonic GABAergic inhibition which could have a profound global effect on sympathetic outflow. Recording from identified SPNs in the intermediolateral cell column (IML) of rat spinal cord slices, application of the GABA receptor antagonist bicuculline, but not gabazine, elicited a change in voltage that lasted for the duration of application. This response was mediated by a direct effect on SPNs since it persisted in tetrodotoxin and low Ca(2+)/high Mg(2+) and the amplitude of responses were related to Cl(-) concentration in patch solutions. Such tonic inhibitory responses were not observed in interneurons, the other neuronal type in the IML, although ongoing IPSPs were antagonized in these neurons. The effects of bicuculline were enhanced by diazepam but not zolpidem or the GABA modulators THIP and THDOC suggesting a role for alpha5 subunits. PCR using primers for the alpha5 and delta subunits indicated the presence of alpha5, but not delta subunits in the IML. Firing rates of SPNs were enhanced by bicuculline and decreased by diazepam indicating that this tonic inhibition has a profound effect on the excitability of SPNs. These data indicate a novel influence for controlling the activity of SPNs regardless of their function.

Topics: Anesthetics; Animals; Animals, Newborn; Autonomic Fibers, Preganglionic; Desoxycorticosterone; Dose-Response Relationship, Drug; Dose-Response Relationship, Radiation; Electric Stimulation; Female; GABA Agents; gamma-Aminobutyric Acid; Glycine Agents; In Vitro Techniques; Male; Membrane Potentials; Neural Inhibition; Neurons; Patch-Clamp Techniques; Phosphinic Acids; Propanolamines; Rats; Sodium Channel Blockers; Strychnine; Tetrodotoxin

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.

Topics: Anesthetics; Animals; Bicuculline; Desoxycorticosterone; Female; GABA Antagonists; GABA Modulators; Glycine Agents; Inhibitory Postsynaptic Potentials; Male; Membrane Potentials; Neural Inhibition; Posterior Horn Cells; Pregnanolone; Rats; Rats, Sprague-Dawley; Receptors, GABA-A; Receptors, Glycine; Strychnine; Synaptic Transmission