COCAINE EFFECTS IN REAGGREGATE TISSUE CULTURE

可卡因对重新聚集组织培养的影响

• 批准号: 3853730
• 负责人: BRUCE D PERRY
• 金额: --
• 依托单位: UNIVERSITY OF CHICAGO
• 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/3853730
• 关键词: adenylate cyclase; alternatives to animals in research; brain cell; cocaine; developmental neurobiology; dopamine; dopamine receptor; dosage; drug abuse; drug metabolism; embryo /fetus cell /tissue; gene expression; neurotransmitter metabolism; psychological reinforcement; psychopharmacology; receptor coupling; tissue /cell culture

Cocaine is a naturally occurring alkaloid with psychomotor stimulant and local anesthetic properties. Due to its strongly reinforcing properties in man it is widely abused in this country. Cocaine abuse has become a major public health problem. The mechanisms underlying cocaine's reinforcing properties are not well understood but appear to involve central dopamine neurons and, via dopamine receptors, their projection fields. Studying the biochemical mechanisms of action of cocaine in animals has been hindered by the complex nature of the intact brain systems including the multiple dopamine systems in the CNS. In intact animal systems, however, it is clear that chronic cocaine alters dopaminergic functioning. The present studies will directly examine cellular mechanisms by which cocaine interacts with CNS dopamine containing neurons in a novel isolated system. Re-aggregate tissue cultures will be used which contain neurochemically-defined cells from fetal mouse brain which have been dissected, dissociated and recombined to form highly organized structures in which innervating and target cells make appropriate functional connections. The advantage of this re-aggregating tissue system is that cells from discrete areas of the brain can be cultured either alone or in combination with the appropriate targets and in the presence and absence of appropriate innervation. Two major sets of experiments will be performed in this model system. The first set will examine the effects of various cocaine exposure paradigms (acute and chronic) on the biochemical pharmacology (e.g., dopamine release, dopamine turnover, dopamine receptor/effecter functioning) of the dopamine neurons in the reaggregate systems (i.e., aggregates with and without appropriate targets). The second set of experiments will take advantage of the inherent developmental nature of the reaggregate systems. The effects of cocaine exposure during development of the reaggregates systems on the phenotypic expression of D1 and D2 receptors and their coupling to effector systems (e.g., adenylate cyclase) will be examined. Issues of tissue specificity, dose effects, and 'critical periods' will be studied. The results of these experiments will enhance significantly the understanding of the mechanisms of action of cocaine in mature, and in developing, dopamine-containing neuronal systems in the brain and may provide important directions for examining cocaine effects in more complex systems.

可卡因是一种天然存在的生物碱，具有精神运动兴奋剂和 局部麻醉特性。由于其强大的增强性能 伙计，它在这个国家被广泛滥用。可卡因滥用已成为主要问题 公共卫生问题。可卡因增强作用的机制 其特性尚不清楚，但似乎涉及中枢多巴胺 神经元以及通过多巴胺受体的投射场。 研究可卡因在动物体内作用的生化机制 受到完整大脑系统复杂性的阻碍，包括 中枢神经系统中的多个多巴胺系统。在完整的动物系统中， 然而，很明显，长期服用可卡因会改变多巴胺能功能。 目前的研究将直接检查细胞机制 可卡因与中枢神经系统中含有多巴胺的神经元相互作用 系统。将使用重新聚集的组织培养物，其中含有 来自胎儿小鼠大脑的神经化学定义的细胞 解剖、分离和重组以形成高度组织的结构 其中神经支配细胞和靶细胞发挥适当的功能 连接。这种重新聚集组织系统的优点是 来自大脑离散区域的细胞可以单独培养，也可以在 与适当的目标相结合，并在存在或不存在的情况下 适当的神经支配。 在此模型系统中将进行两组主要实验。这 第一组将检查各种可卡因暴露范例的影响 （急性和慢性）生化药理学（例如多巴胺 释放、多巴胺周转、多巴胺受体/效应器功能） 重新聚集系统中的多巴胺神经元（即与 和 聚集） 没有适当的目标）。第二组实验将进行 重新聚合系统固有的发展性质的优势。 重新聚集体发育过程中可卡因暴露的影响 D1 和 D2 受体表型表达系统及其影响 将检查与效应系统（例如腺苷酸环化酶）的耦合。 组织特异性、剂量效应和“关键期”的问题将是 研究过。这些实验的结果将显着提高 了解可卡因在成熟和中的作用机制 大脑中正在发育的含多巴胺的神经元系统，可能 为检查更复杂的可卡因效应提供重要方向 系统。