MEASUREMENT OF RAPID CHANGE IN REGIONAL BRAIN METABOLISM

该提案旨在进一步测试、完善和开发一种程序， 在这个实验室中，使用2-脱氧葡萄糖（2DG）来测量短期 大脑不同部位葡萄糖代谢率的变化。的 众所周知的Sokoloff的2DG方法测量葡萄糖代谢率 通过14 C-2DG-6-磷酸盐（DG 6P）蓄积放射自显影 在45分钟的时间段内。这一漫长的时期主要是为了 消散2DG，这在放射自显影中是无法区分的 从它的磷酸盐代谢物中。因为大脑中的许多有趣的事件 在更短的时间内发生，45分钟的延迟代表了 原始2DG方法的明显局限性。 新方法使用非放射性2DG， 放射自显影，DG 6P、2DG和葡萄糖的直接酶测定 在大脑的相关区域。短至一分钟的时间是实用的 分析小到大到胞体的脑区都是可行的。 不需要对这三种化合物进行物理分离。的测定 基于区分DG 6P和G6 P的酶方法， 能够根据需要进行几乎无限的放大。

The proposal is to further test, refine, and exploit a procedure developed in this laboratory which uses 2-deoxyglucose (2DG) to measure short-term changes in glucose metabolic rates in different parts of the brain. The well known 2DG method of Sokoloff measures glucose metabolic rate radioautographically by the accumulation of 14C-2DG-6-phosphate (DG6P) during a 45 min time period. This long period is required largely to dissipate the 2DG, which would be indistinguishable in the radioautograph from its phosphate metabolite. Because many events of interest in brain occur within a much shorter time frame, the 45 min delay represents a distinct limitation on the original 2DG method. The new method uses non-radioactive 2DG and substitutes for radioautography, the direct enzymatic measurement of DG6P, 2DG, and glucose in the brain areas of interest. Times as short as one minute are practical and analyses of brain areas as small as large perikarya are feasible. Physical separations of the three compounds are not required. The assays are based on enzymatic methods which distinguish DG6P from G6P, and which are capable of virtually unlimited amplification, according to need.