ETHANOL TOLERANCE AND DEPENDENCE IN VITRO

体外乙醇耐受性和依赖性

基本信息

批准号：
3108911
负责人：
ELLIOTT RICHELSON
金额：
$ 14.19万
依托单位：
MAYO CLINIC ROCHESTER
依托单位国家：
美国
项目类别：
财政年份：
1980
资助国家：
美国
起止时间：
1980-04-01 至 1989-06-30
项目状态：
已结题

项目摘要

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.

乙醇的药理学和成瘾性作用的基因座可能是神经递质与其受体之间的相互作用水平突触。因此，该提案的重点是急性和慢性乙醇在具有环状的某些受体的功能上 AMP和环状GMP作为推定的第二使者。培养的人皮与前列腺素和β-肾上腺素能受体介导的成纤维细胞环状AMP合成和鼠神经母细胞瘤细胞（克隆N1E-115）几种受体（毒蕈碱，组胺H1，血管紧张素11，Bradyinin，Bradyinin，神经素和凝血酶）介导环状GMP合成被用作这项工作的模型系统。人类皮肤成纤维细胞的研究将确定在动物研究中是否发现结果（包括我们与鼠神经母细胞瘤细胞）适用于人体受体。研究与鼠神经母细胞瘤细胞一起可能有助于阐明机制乙醇耗尽了环状GMP的大脑水平。前列腺素和人皮肤成纤维细胞的β-肾上腺素能受体将彻底以生物测定（环状AMP产生）和在研究乙醇之前，放射性结合测定法。循环 GMP对神经母细胞瘤细胞的研究将在我们的数据中跟进乙醇的混合抑制类型（竞争性和非竞争性）受体介导的环状GMP合成，并适应了这种影响细胞长期暴露于这种酒精时，乙醇。自从受体介导的环状GMP合成涉及磷脂，乙醇对肌醇磷酸盐和释放的影响将研究蛛网酸。