Volatile general anaesthetic actions on recombinant nACh(α7), 5-HT(3) and chimeric nACh(α7)-5-HT(3) receptors expressed in Xenopus oocytes

The effect of halothane and isoflurane was studied on the function of recombinant neurotransmitter receptors expressed in Xenopus oocytes. Both anaesthetics inhibited nicotinic acetylcholine type α7 (nACh(α7)) receptor-mediated responses, potentiated 5-hydroxytryptamine type 3 (5-HT(3)) receptor-mediated responses at low agonist concentrations, and inhibited the function of a chimeric receptor (with the N-terminal domain from the nACh(α7) receptor and the transmembrane and C-terminal domains from the 5-HT(3) receptor) in a manner similar to that of the nACh(α7) receptor. Since the N-terminal domain of the chimeric receptor was from the nACh(α7) receptor, the observations suggest that the inhibition involves the N-terminal domain of the receptor

Inhibition of excitatory amino acid-activated currents by trichloroethanol and trifluoroethanol in mouse hippocampal neurones

1. The effects of the active metabolite of chloral derivative sedative-hypnotic agents, 2,2,2-trichloroethanol (trichloroethanol), and its analog 2,2,2-trifluoroethanol (trifluoroethanol), were studied on ion current activated by the excitatory amino acids N-methyl-D-aspartate (NMDA) and kainate in mouse hippocampal neurones in culture using whole-cell patch-clamp recording. 2. Both trichloroethanol and trifluoroethanol inhibited excitatory amino acid-activated currents in a concentration-dependent manner. Trichloroethanol inhibited NMDA- and kainate-activated currents with IC(50) values of 6.4 and 12 mM, respectively, while trifluoroethanol inhibited NMDA- and kainate-activated currents with IC(50) values of 28 and 35 mM, respectively. 3. Both trichloroethanol and trifluoroethanol appeared to be able to inhibit excitatory amino acid-activated currents by 100 per cent. 4. Concentration-response analysis of NMDA- and kainate-activated current revealed that trichloro-ethanol decreased the maximal response to both agonists without significantly affecting their EC(50) values. 5. Both trichloroethanol and trifluoroethanol inhibited excitatory amino acid-activated currents more potently than did ethanol. The inhibitory potency of trichloroethanol and trifluoroethanol appears to be associated with their increased hydrophobicity. 6. The observation that trichloroethanol inhibits excitatory amino acid-activated currents at anaesthetic concentrations suggests that inhibition of excitatory amino acid receptors may contribute to the CNS depressant effects of chloral derivative sedative-hypnotic agents

Inhibition by ethanol of rat P2X(4) receptors expressed in Xenopus oocytes

1. The effect of ethanol on the function of P2X(4) receptors expressed in Xenopus oocytes was studied using two-electrode voltage-clamp recording. 2. The amplitude of current activated by 1 μM ATP was decreased by ethanol in a concentration-dependent manner over the concentration range 1–500 mM. The concentration of ethanol that produced 50% inhibition (IC(50)) of current activated by 1 μM ATP was 58 mM. 3. Ethanol inhibition of ATP-activated current was not dependent on membrane potential from −60 to +20 mV, and ethanol did not change the reversal potential of ATP-activated current. 4. Ethanol, 50 mM, shifted the ATP concentration-response curve to the right, increasing the EC(50) for ATP from 9.1 to 16.0 μM, but did not reduce the maximal response to ATP. 5. The results suggest that ethanol may inhibit P2X(4) receptors by decreasing the apparent affinity of the binding site for ATP. 6. Since the P2X(4) receptor is the most abundant P2X subunit in the brain, these receptors could be important effectors of ethanol action in the central nervous system

The mechanism by which ethanol inhibits rat P2X(4) receptors is altered by mutation of histidine 241

1. We investigated ethanol inhibition of the rat P2X(4) receptor and the contribution of the three histidine residues in the extracellular loop of this receptor to ethanol inhibition of receptor function, using site-directed mutagenesis and electrophysiological characterization of recombinant receptors. 2. In the wild-type receptor, 50, 200 and 500 mM ethanol increasingly shifted the ATP concentration–response curve to the right in a parallel manner, increasing the EC(50) value without affecting E(max). However, 750 or 900 mM ethanol did not produce a further increase in the EC(50) value of the ATP concentration–response curve, suggesting that this inhibition is not competitive. 3. The P2X(4) receptor mutations H140A and H286A did not significantly alter ethanol inhibition of ATP-activated current. By contrast, the mutation H241A changed the mechanism by which ethanol inhibits receptor function; viz., ethanol inhibition was not associated with an increased EC(50) value of the ATP concentration–response curve, instead, ethanol decreased the maximal response to ATP without affecting the EC(50) value of the ATP concentration–response curve. 4. Ethanol inhibition of the H241A mutant was voltage independent between −60 and +20 mV and ethanol did not alter the reversal potential of ATP-activated current. In addition, ethanol decreased the desensitization rate of the H241A-mediated current. 5. The purinoceptor antagonists, suramin and pyridoxal-phosphate-6-azophenyl-2′,4′-disulphonic acid (PPADS), did not alter the magnitude of ethanol inhibition of ATP-activated current in the H241A mutant. 6. The results suggest that ethanol inhibits the wild-type rat P2X(4) receptor by an allosteric action to increase the EC(50) value of the ATP concentration–response curve, the P2X(4) receptor mutation H241A alters the mechanism by which ethanol inhibits P2X(4) receptor function, and ethanol and PPADS or suramin appear to inhibit H241A-mutated receptors at independent sites