Harris R A, Baxter D M, Mitchell M A, Hitzemann R J
Mol Pharmacol. 1984 May;25(3):401-9.
DBA/2 mice were made tolerant to and dependent on ethanol by administration of an ethanol-containing liquid diet for 7 days. Fluorescent probe molecules were used to estimate the fluidity and ethanol sensitivity of brain synaptic membranes from these mice. The fluorescence polarization of cis- parinarate , trans- parinarate , and 1,6-diphenyl-1,3,5-hexatriene (probes of the membrane core) and 1-(4- trimethylammoniumphenyl )-6-phenyl-1,3,5-hexatriene (a probe of the membrane surface) was higher in membranes from ethanol tolerant-dependent mice than in membranes from control mice. The decrease in fluorescence polarization produced by in vitro exposure to ethanol was attenuated in membranes from ethanol tolerant-dependent mice when 1,6-diphenyl-1,3,5-hexatriene was used as the probe, but not when 1-(4- trimethylammoniumphenyl )-6-phenyl-1,3,5-hexatriene was used. These results indicate that chronic ingestion of ethanol decreased the fluidity and the ethanol sensitivity of the synaptic membranes. In contrast to the alterations observed with intact membranes, liposomes of lipids extracted from synaptic membranes of control and ethanol tolerant-dependent mice did not differ in their physical properties. Analysis of membrane lipids demonstrated that chronic ethanol treatment selectively decreased the unsaturated acyl groups of phosphatidylserine without altering the acyl composition of other phospholipids or sphingolipids. The amount of each phospholipid was not changed, but membrane cholesterol was decreased by chronic ethanol ingestion. Use of 2-dimensional thin-layer chromatography allowed the quantitation of 10 different gangliosides. The concentrations of these lipids were unchanged in synaptic membranes from ethanol tolerant-dependent mice. Thus, the changes in membrane physical properties produced by chronic ingestion of ethanol may be due, at least in part, to altered acyl composition of phosphatidylserine. The differences observed between intact membranes and extracted lipids suggest, however, that chronic ethanol treatment also produced changes in the lipid arrangement or lipid-protein interactions of the intact membranes.
通过给予含乙醇的液体饮食7天,使DBA/2小鼠对乙醇产生耐受性并产生依赖。使用荧光探针分子来评估这些小鼠脑突触膜的流动性和乙醇敏感性。顺式-紫黄质、反式-紫黄质和1,6-二苯基-1,3,5-己三烯(膜核心探针)以及1-(4-三甲基铵苯基)-6-苯基-1,3,5-己三烯(膜表面探针)在乙醇耐受依赖小鼠的膜中的荧光偏振度高于对照小鼠膜中的荧光偏振度。当使用1,6-二苯基-1,3,5-己三烯作为探针时,乙醇耐受依赖小鼠膜中体外暴露于乙醇引起的荧光偏振度降低减弱,但使用1-(4-三甲基铵苯基)-6-苯基-1,3,5-己三烯时则不然。这些结果表明,长期摄入乙醇会降低突触膜的流动性和乙醇敏感性。与完整膜观察到的变化相反,从对照和乙醇耐受依赖小鼠的突触膜中提取的脂质脂质体在物理性质上没有差异。膜脂质分析表明,长期乙醇处理选择性降低了磷脂酰丝氨酸的不饱和酰基,而不改变其他磷脂或鞘脂的酰基组成。每种磷脂的量没有变化,但长期摄入乙醇会降低膜胆固醇。二维薄层色谱法的使用使得能够定量10种不同的神经节苷脂。这些脂质的浓度在乙醇耐受依赖小鼠的突触膜中没有变化。因此,长期摄入乙醇引起的膜物理性质变化可能至少部分归因于磷脂酰丝氨酸酰基组成的改变。然而,完整膜和提取脂质之间观察到的差异表明,长期乙醇处理也会使完整膜的脂质排列或脂质-蛋白质相互作用发生变化。