Smith Sheryl S, Gong Qi Hua
Department of Physiology and Pharmacology, Box 31, SUNY Downstate Medical Center, 450 Clarkson Avenue, Brooklyn, NY 11203, USA.
Alcohol. 2007 May;41(3):223-31. doi: 10.1016/j.alcohol.2007.04.007.
Several recent studies have suggested that alphabetadelta subtypes of gamma-aminobutyric acid type A (GABAA) receptors (delta-GABAR) are a target for low dose ethanol (<30 mM). However, there are also conflicting reports suggesting that only high doses of the drug (100 mM) modulate these receptors. In addition, the studies which have demonstrated a clear effect of low dose ethanol on delta-GABAR find different effective concentrations for this effect. Here, we test the hypothesis that the apparent disparity in effective concentration is due to time-course effects when low (1-3 mM) dose ethanol is preapplied. To this end, we tested ethanol effects on native GABAR in CA1 hippocampus in a model of increased alpha4betadelta GABAR expression following 48h administration of the GABA-modulatory steroid THP (3alpha-OH-5beta-pregnan-20-one) to adult, female rats. GABA(EC20)-gated current was recorded with whole-cell patch clamp procedures from acutely isolated pyramidal cells. We assessed ethanol's effect on GABA-gated current using either (1) 2-5 min application of ethanol in increasing concentrations (0.1-30 mM) or (2) coadministration of ethanol with GABA. Two minute application of 1-3 mM ethanol produced optimal potentiation of GABA-gated current following steroid treatment, with higher concentrations less effective. In contrast, 30 mM ethanol produced optimal effects when ethanol was not preapplied. However, following preapplication of 1mM ethanol, 30 mM ethanol decreased the peak GABA-gated current. These findings suggest that ethanol may act at multiple interacting sites to affect GABAR efficacy and desensitization. These data also suggest that ethanol effects on GABA-gated current are affected by the time course of exposure and previous exposure to low concentrations of the drug.
最近的几项研究表明,A型γ-氨基丁酸(GABAA)受体的αδ亚型(δ-GABAR)是低剂量乙醇(<30 mM)的作用靶点。然而,也有相互矛盾的报道表明,只有高剂量的药物(100 mM)才能调节这些受体。此外,那些已证明低剂量乙醇对δ-GABAR有明显作用的研究,发现该作用的有效浓度各不相同。在此,我们检验以下假设:当预先施加低剂量(1-3 mM)乙醇时,有效浓度的明显差异是由于时间进程效应所致。为此,我们在成年雌性大鼠经48小时给予GABA调节性类固醇THP(3α-羟基-5β-孕烷-20-酮)后,α4βδ GABAR表达增加的模型中,测试了乙醇对CA1海马区天然GABAR的影响。采用全细胞膜片钳技术,从急性分离的锥体细胞中记录GABA(EC20)门控电流。我们使用以下两种方法评估乙醇对GABA门控电流的影响:(1)以递增浓度(0.1-30 mM)施加乙醇2-5分钟;(2)将乙醇与GABA共同给药。在类固醇处理后,施加2分钟的1-3 mM乙醇可使GABA门控电流产生最佳增强作用,浓度越高效果越差。相比之下,当未预先施加乙醇时,30 mM乙醇产生最佳效果。然而,在预先施加1 mM乙醇后,30 mM乙醇会降低GABA门控电流的峰值。这些发现表明,乙醇可能作用于多个相互作用位点,以影响GABAR的功效和脱敏作用。这些数据还表明,乙醇对GABA门控电流的影响受暴露时间进程以及先前暴露于低浓度药物的影响。