RESUMO
Glycine receptor function mediates most inhibitory neurotransmission in the brainstem and spinal cord and is enhanced by alcohols, volatile anesthetics, inhaled drugs of abuse, and endogenous compounds including zinc. Because zinc exists ubiquitously throughout the brain, investigations of its effects on the enhancement of GlyR function by alcohols and anesthetics are important to understanding the effects of these agents in vivo. In the present study, the effects of zinc plus ethanol, pentanol, or isoflurane were tested on homomeric alpha1 glycine receptors to determine if concurrent applications of physiological concentrations of zinc with each of these modulators changed the magnitude of their effects. Homomeric alpha1 glycine receptors were expressed in Xenopus laevis oocytes, and the two-electrode voltage-clamp technique was used to measure glycine-mediated currents in the presence of combinations of zinc with ethanol, pentanol or isoflurane. The combined effects of zinc plus ethanol were greater than the sum of the effects produced by either compound alone. However, this was not seen when zinc was combined with either pentanol or isoflurane. Chelation of zinc by tricine decreased the effects of sub-maximal, but not maximal, concentrations of glycine, and diminished the magnitude of ethanol enhancement observed. These findings suggest a zinc/ethanol interaction at the alpha1 GlyR that results in the enhancement of the effects of ethanol action on GlyR function.
Assuntos
Depressores do Sistema Nervoso Central/farmacologia , Etanol/farmacologia , Potenciais da Membrana/fisiologia , Receptores de Glicina/metabolismo , Zinco/farmacologia , Análise de Variância , Animais , Relação Dose-Resposta a Droga , Interações Medicamentosas , Estimulação Elétrica/métodos , Feminino , Glicina/análogos & derivados , Glicina/farmacologia , Potenciais da Membrana/efeitos dos fármacos , Potenciais da Membrana/genética , Microinjeções/métodos , Oócitos , Técnicas de Patch-Clamp , Receptores de Glicina/genética , Análise Espectral/métodos , XenopusRESUMO
The glycine receptor (GlyR) is a ligand-gated ion channel and member of the nicotinic acetylcholine receptor superfamily. Acting as allosteric modulators of receptor function, drugs such as alcohol and volatile anesthetics enhance the function of GlyRs. The actions of these drugs at inhibitory receptors in the brain and spinal cord are thought to produce many of the physiological effects associated with their use. The actions of ethanol on the GlyR have been well studied on the macroscopic, whole cell level. We examined the effects of 3 microM glycine +/- 50 or 200 mM ethanol on outside-out patches pulled from Xenopus laevis oocytes expressing wild-type alpha1 GlyR, to determine the effects of alcohol at the single-channel level. Alcohol enhanced GlyR function in a very specific manner. It had minimal effects on open and closed dwell times and likelihood. Instead, ethanol potentiated GlyR function almost exclusively by increasing burst durations and increasing the number of channel openings per burst, without affecting the percentage of open time within bursts. Kinetic modeling suggests that ethanol increases burst durations by decreasing the rate of glycine unbinding.