Quantitative analysis of Pharmacological effects of psychiatic drugs using neuronal cultured cells

利用神经元培养细胞定量分析精神药物的药理作用

基本信息

批准号：
02671049
负责人：
SAWADA Yasufumi
金额：
$ 1.41万
依托单位：
University of Tokyo
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1990
资助国家：
日本
起止时间：
1990 至 1991
项目状态：
已结题

项目摘要

In this study, we were successful in obtaining mouse cerebellar granule cells in monolayer culture. They were GABA neuron because we found out specific binding of ^3H-mucizol to them. In the presence time, however, we were unsuccessful in measuring the glucose utilization by using these cells. Therefore, we measured the glucose utilization by using isolated brain cells and ^<14>C-2-deoxyglucose.Furthermore, the effect of muscimol, a GABAA receptor agonist, on the basic metabolic activity was investigated in the mouse barin and was correlated with its receptor occupancy. For the quantitative evaluation of the functional activity of the mouse brain, the cerebral glucose utilization was measured by the double tracer technique, using ^<14>C-2-deoxyglucose and ^3H-3-O-methylglvcose. The dose-dependent reduction in the cerebral glucose utilization was observed after i. v. administration of various doses of muscizol. O. n the other hand, the GABAA receptor occupancy of muscimol was determined by using the values of the unbound drug concentration in the brain tissue and the receptor dissociation constant based on the in vitro binding experiments use in the dissociated brain cells. The tissue unbound concentration of muscimol was calculated by multiplying the total concentration in the brain after administration of muscimol and the tissue unbound fraction, which was measured by the equilibrium dialysis method using brain tissue homogenate. A linear relationship was observed between the GABAA receptor occupancy of muscimol and the decrease in the cerebral glucose utilization. This finding indicates that the simple receptor occupancy theory holds for this receptorligand system and there is a large difference in the effect of glucose metabolic response between GABA_A receptor-agonist interaction and benzodiazepine receptor-agonist interaction.

在这项研究中，我们成功地获得了单层培养的小鼠小脑颗粒细胞。它们是 GABA 神经元，因为我们发现 ^3H-mucizol 与它们特异性结合。然而，在存在时间内，我们未能成功地使用这些细胞测量葡萄糖的利用率。因此，我们通过使用分离的脑细胞和^ 14 C-2-脱氧葡萄糖来测量葡萄糖利用。此外，在小鼠巴林中研究了蝇蕈醇(GABAA受体激动剂)对基本代谢活动的影响，并将其与它的受体占据。为了定量评价小鼠脑的功能活动，通过双示踪技术使用14C-2-脱氧葡萄糖和3H-3-O-甲基葡萄糖测量脑葡萄糖利用率。 i后观察到脑葡萄糖利用率的剂量依赖性降低。 v.给予不同剂量的muscizol。另一方面，通过使用脑组织中未结合药物浓度的值和基于在离解脑细胞中使用的体外结合实验的受体解离常数来确定蝇蕈醇的GABAA受体占据。蝇蕈醇的组织非结合浓度通过将给予蝇蕈醇后脑中的总浓度乘以组织非结合分数来计算，该组织非结合分数是通过使用脑组织匀浆的平衡透析法测量的。观察到蝇蕈醇的 GABAA 受体占据与脑葡萄糖利用率的降低之间存在线性关系。这一发现表明简单的受体占据理论适用于该受体配体系统，并且GABA_A受体-激动剂相互作用和苯二氮卓类受体-激动剂相互作用之间的葡萄糖代谢反应的效果存在很大差异。