ETHANOL TOLERANCE AND DEPENDENCE IN VITRO

乙醇的药理和成瘾作用的中心可能在 脑内神经递质与其受体相互作用的水平 Synapse。因此，本提案侧重于急性和非传染性疾病的影响 慢性乙醇对某些受体功能的影响 AMP和环状GMP可能是第二信使。培养的人皮肤 前列腺素和β-肾上腺素能受体介导的成纤维细胞 环磷酸腺苷合成与小鼠神经母细胞瘤细胞(克隆N1E-115) 几种受体(毒鼠碱、组胺H1、血管紧张素11、缓激肽、 神经降压素和凝血酶)介导的循环GMP合成被用作 这项工作的模型系统。对人类皮肤成纤维细胞的研究将 确定在动物研究中发现的结果(包括我们在 小鼠神经母细胞瘤细胞)适用于人类受体。研究 与小鼠神经母细胞瘤细胞的关系可能有助于阐明 酒精会消耗大脑中循环GMP的水平。前列腺素和 人皮肤成纤维细胞的β-肾上腺素能受体将被彻底 以生物测定(环化AMP生产)和 乙醇研究前的放射性配基结合分析。The Cycle 神经母细胞瘤细胞的GMP研究将对我们的数据进行后续研究，显示 乙醇的混合型抑制(竞争性和非竞争性) 受体介导的环状GMP合成及其对这一效应的适应 乙醇使细胞长期暴露在酒精中。自.以来 受体介导的环状GMP合成涉及细胞周转 磷脂，乙醇对肌醇磷酸和肌醇释放的影响 将对花生四烯酸进行研究。

The locus of ethanol's pharmacological and addictive effects may be at the level of the interaction between a neurotransmitter and its receptor in the synapse. Therefore this proposal focuses on the effects of acute and chronic ethanol on the functioning of certain receptors which have cyclic AMP and cyclic GMP as putative second messengers. Cultured human skin fibroblasts with prostaglandin and Beta-adrenergic receptors mediating cyclic AMP synthesis and murine neuroblastoma cells (clone N1E-115) with several receptors (muscarinic, histamine H1, angiotensin 11, bradykinin, neurotensin and thrombin) mediating cyclic GMP synthesis are being used as model systems for this work. Studies with human skin fibroblasts will determine whether results found in animal studies (including our work with murine neuroblastoma cells) are applicable to human receptors. Studies with murine neuroblastoma cells may help elucidate the mechanism whereby ethanol depletes brain levels of cyclic GMP. The prostaglandin and Beta-adrenergic receptors of human skin fibroblasts will be thoroughly characterized by biological assay (cyclic AMP production) and by radioligand binding assay prior to the studies with ethanol. The cyclic GMP studies with neuroblastoma cells will follow-up on our data showing a mixed type of inhibition (competitive and noncompetitive) by ethanol of receptor-mediated cyclic GMP synthesis and an adaptation to this effect of ethanol upon chronic exposure of cells to this alcohol. Since receptor-mediated cyclic GMP synthesis involves the turnover of phospholipids, effects of ethanol on the release of inositol phosphates and arachidonate acid will be studied.