REGULATION OF SECOND MESSENGER SYSTEMS IN EBV-TRANSFORMED B-CELL LINES

EBV 转化的 B 细胞系中第二信使系统的调节

• 批准号: 3781514
• 负责人: J M MASSERANO
• 金额: --
• 依托单位: NATIONAL INSTITUTE OF MENTAL HEALTH
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/3781514
• 关键词: B lymphocyte; Epstein Barr virus; biological models; cell transformation; cholera toxin; cyclic AMP; dopamine agonists; enzyme inhibitors; forskolin; human tissue; isoproterenol; phorbols; postmortem; prostaglandin E; protein kinase C; radioimmunoassay; schizophrenia; second messengers; tissue /cell culture

Schizophrenia is an idiopathic and probably heterogenous disorder. Evidence suggest that disturbances in the cyclic AMP second messenger system may contribute to the pathophysiology of schizophrenia. Studies using postmortem brain tissue have shown enhanced cAMP production in response to NaF, Gpp[NH]p and a D1 receptor agonist in schizophrenic patients as compared to normals (Science 221:1304, 1983). Similar changes have been reported in schizophrenic patients when cyclic AMP is evaluated in a readily accessible source of tissue: blood cells. Our interest in the cyclic AMP second messenger system relates to the fact that catecholaminergic neurons in the central nervous system produce their effects predominantly through activation of second messenger systems. The second messenger, cyclic AMP, is regulated intracellularly by dopamine (D1, D2), norepinephrine (beta1, alpha2) and epinephrine (beta1, beta2, alpha2). In order to study cyclic AMP production in schizophrenic patients as compared to controls, we have initiated studies using beta-lymphocytes that have been transformed with the Epstein-Barr virus. In our group, Dr. Kulaga has transformed cells from twins discordant for schizophrenia, and from unrelated schizophrenic patients and normals. We feel that the availability of these cells offers an ideal opportunity for evaluating, under a controlled setting, the regulation of second messenger systems in schizophrenia patients. Initial studies are in process to evaluate compounds that regulate cyclic AMP accumulation in these transformed B cells. Cells were retreated with and without the protein kinase C activator, PMA, followed by treatment with forskolin, cholera toxin, PGE1, isoproterenol and dopamine agonists. The cyclic AMP production in response to these agents was measured using RIA. Forskolin, cholera toxin, PGE 1, PMA, dopamine and isoproterenol increased cyclic AMP accumulation in these cells. Pretreatment of the cells with PMA potentiated the cyclic AMP accumulation produced by forskolin, cholera toxin, pGE1, isoproterenol and dopamine. The effects of PMA were partially antagonized by staurosporin, an inhibitor of protein kinase C. These preliminary results indicate that the B cell lines are a useful model to study the regulation of cyclic AMP in human cells.

精神分裂症是一种特发性的，可能是异质性的疾病。 有证据表明，环磷酸腺苷第二信使的干扰 系统可能有助于精神分裂症的病理生理学。 研究 使用死后脑组织的研究显示， 精神分裂症患者对NaF、Gpp[NH]p和D1受体激动剂的反应 患者与正常人相比（Science 221：1304，1983）。 类似的变化 在精神分裂症患者中， 在一个容易获取的组织来源：血细胞。 我们关心 环腺苷酸第二信使系统涉及的事实， 中枢神经系统中的儿茶酚胺能神经元产生它们的 主要通过激活第二信使系统发挥作用。 的 第二信使环AMP在细胞内受多巴胺调节（D1， D2）、去甲肾上腺素（β 1，α 2）和肾上腺素（β 1，β 2，α 2）。 为了研究精神分裂症患者的环磷酸腺苷产生， 与对照组相比，我们已经开始使用β-淋巴细胞进行研究， 已经被EB病毒转化了 在我们的团队中，博士。 Kulaga已经转化了来自精神分裂症不和谐双胞胎的细胞， 无关的精神分裂症患者和正常人 我们觉得 这些电池的可用性提供了一个理想的评估机会， 在受控环境下， 精神分裂症患者 正在进行初步研究， 调节这些转化的B中环AMP积累的化合物 细胞 用蛋白激酶C和不用蛋白激酶C对细胞进行再处理 激活剂，PMA，然后用毛喉素，霍乱毒素，PGE 1， 异丙肾上腺素和多巴胺激动剂。 环磷酸腺苷的产生 用放射免疫分析法测定了这些药物的敏感性。 毛喉素，霍乱毒素，前列腺素E1， PMA，多巴胺和异丙肾上腺素增加环磷酸腺苷的积累， 细胞 PMA预处理细胞可增强cAMP的生成 毛喉素、霍乱毒素、pGE 1、异丙肾上腺素和 多巴胺 PMA的作用可被星形孢菌素部分拮抗， 蛋白激酶C的抑制剂。 这些初步结果表明， B细胞系是研究cAMP调节的有用模型 在人类细胞中。