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

本研究成功地获得了小鼠小脑颗粒细胞的单层培养。由于发现了^3H-粘菌唑与它们的特异性结合，因此它们是GABA神经元。然而，在存在时间内，我们通过使用这些细胞测量葡萄糖利用率是不成功的。因此，我们用离体脑细胞和~（12）C-2-脱氧葡萄糖测定了葡萄糖的利用率<14>，并进一步研究了蝇蕈醇（GABAA受体激动剂）对小鼠barin基础代谢活动的影响，并将其与受体结合率进行了相关性研究。为了定量评价小鼠脑的功能活动，用双示踪技术，使用^<14>C-2-脱氧葡萄糖和^3 H-3-O-甲基葡萄糖，测量了脑葡萄糖利用率。脑葡萄糖利用率呈剂量依赖性降低。V.施用不同剂量的蝇蕈唑。O.另一方面，蝇蕈醇的GABAA受体占有率通过使用脑组织中的未结合药物浓度和基于在解离的脑细胞中使用的体外结合实验的受体解离常数的值来确定。通过将蝇蕈醇给药后脑中的总浓度乘以组织未结合分数来计算蝇蕈醇的组织未结合浓度，所述组织未结合分数通过平衡透析法使用脑组织匀浆测量。蝇蕈醇的GABAA受体占有率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，简单受体占据理论适用于这种受体-配体系统，GABA_A受体-激动剂相互作用对葡萄糖代谢反应的影响与苯二氮卓类受体-激动剂相互作用的影响有很大差异。

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.