ETHANOL TOLERANCE AND DEPENDENCE IN VITRO

拟议研究的总体目标是确定急性和 乙醇对神经递质受体功能和膜的慢性效应 流动性 含前列腺素的小鼠神经母细胞瘤（N1 E-115）的克隆 和毒蕈碱乙酰胆碱受体以及大鼠胶质瘤（C-6）的克隆， 将在这些研究中使用含有β-肾上腺素能受体的药物。 这些 受体将通过药理学和放射性配体结合技术进行测定。 受体介导的环AMP（前列腺素，β-肾上腺素能受体）和 环GMP（毒蕈碱受体）的合成将被用于 研究这些受体，而（3 H）前列腺素E1，（3 H）奎宁环二苯乙酸， 和（125 I）碘羟苄基吲哚洛尔结合将用于直接测定 前列腺素、毒蕈碱乙酰胆碱和β-肾上腺素能受体， 分别 我们的初步实验表明，乙醇对 前列腺素和毒蕈碱乙酰胆碱受体的功能。 此外，本发明还提供了一种方法， 暴露于乙醇几天后，细胞产生了耐受性， 酒精对这些受体的影响。 我们的一些数据可以解释 如其他人所报道的，乙醇引起的膜流动性的变化。 因此，在本发明中， 荧光偏振技术将用于获得直接证据， 乙醇对这些培养细胞的膜流动性的影响。 的作用 急性和慢性乙醇作用中膜流动性的变化 神经递质受体功能和诱导细胞耐受性 乙醇可以从这些研究中被阐明。

The overall objectives of the proposed research are to determine the acute and chronic effects of ethanol on neurotransmitter receptor function and on membrane fluidity. A clone of mouse neuroblastoma (N1E-115) which contains prostaglandin and muscarinic acetylcholine receptors and a clone of rat glioma (C-6) which contains Beta-adrenergic receptor will be used in these studies. These receptors will be assayed by pharmacological and radioligand binding techniques. Receptor-mediated cyclic AMP (prostaglandin, Beta-adrenergic receptors) and cyclic GMP (muscarinic receptor) synthesis will be used to pharmacologically study these receptors while (3H)prostaglandin E1, (3H)quinuclidinylbenzilate, and (125I)iodohydroxybenzylpindolol binding will be used for direct assay of the prostaglandin, muscarinic acetylcholine and Beta-adrenergic receptors, respectively. Our preliminary experiments showed an effect of ethanol on the function of prostaglandin and muscarinic acetylcholine receptors. In addition, after several days of exposure to ethanol, there developed a cellular tolerance to the effects of ethanol on these receptors. Some of our data may be explained by changes in membrane fluidity caused by ethanol as reported by others. Thus, fluorescence polarization techniques will be used to obtain direct evidence for an effect of ethanol on membrane fluidity of these cultured cells. The role of membrane fluidity changes in the acute and chronic effects of ethanol on neurotransmitter receptor function and in the induction of cellular tolerance to ethanol may be elucidated from these studies.