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采用新型双腔推挽式导管系统对大鼠体内收集的皮质氨基酸神经递质进行体内分析。

In vivo analysis of cortical amino acid neurotransmitters collected in the rat by a new double lumen push-pull catheter system.

作者信息

Peinado J M, Myers R D

机构信息

Department of Pharmacology, East Carolina University School of Medicine, Greenville, NC 27858.

出版信息

Neuroscience. 1988 Mar;24(3):1027-34. doi: 10.1016/0306-4522(88)90084-x.

Abstract

The release of both endogenous and newly synthesized amino acid neurotransmitters was examined simultaneously in different areas of the cerebral cortex in the freely moving rat. An array of push-pull guide tubes was implanted permanently to rest above the frontal, parietal, temporal and occipital areas of the cortex of each rat. Then a new double-lumen catheter system, specially adapted for localized push-pull perfusion of the conscious animal, was used to perfuse an artificial cerebrospinal fluid at each cortical site. For the new synthesis experiments, 0.5 microCi of [14C]glucose in a volume of 2.0 microliter was first microinjected into the perfusion site as a precursor to label amino acids. After the site was perfused at a rate of 12.0 microliter/min, each of the samples was assayed by two-dimensional thin-layer chromatography. In a second analysis, the content of six endogenous amino acids present in unlabeled samples of push-pull perfusate was quantified by high-performance liquid chromatography analysis with electrochemical detection. The results showed a notable homogeneity among each of the four cortical areas in the content of four of the six amino acids examined. Endogenous glutamine exhibited the highest proportional content in the cortical perfusates, whereas glutamic acid was proportionally higher in terms of new synthesis. An anatomical analysis revealed that the level of endogenous glutamic acid in the frontal area was significantly lower than that found in the occipital or temporal regions of the rat's cortex. An opposite result was obtained when the proportional synthesis of glutamic acid from [14C] glucose was compared in different cortical regions in that a statistically higher release occurred in the frontal than in the occipital cortex.

摘要

在自由活动的大鼠大脑皮层的不同区域,同时检测内源性和新合成的氨基酸神经递质的释放情况。将一排推挽式导管理永久性植入每只大鼠皮层的额叶、顶叶、颞叶和枕叶上方。然后,使用一种专门为清醒动物局部推挽灌注设计的新型双腔导管系统,在每个皮层部位灌注人工脑脊液。在新合成实验中,首先将2.0微升体积中含有0.5微居里[14C]葡萄糖的溶液微量注射到灌注部位,作为标记氨基酸的前体。以12.0微升/分钟的速率对该部位进行灌注后,每个样品通过二维薄层色谱法进行分析。在第二次分析中,通过电化学检测的高效液相色谱分析法对推挽灌注液未标记样品中存在的六种内源性氨基酸的含量进行定量。结果显示,在所检测的六种氨基酸中的四种氨基酸含量方面,四个皮层区域中的每一个区域之间都存在显著的同质性。内源性谷氨酰胺在皮层灌注液中的比例含量最高,而谷氨酸在新合成方面的比例更高。解剖学分析表明,额叶区域内源性谷氨酸的水平明显低于大鼠皮层枕叶或颞叶区域内发现的水平。当比较不同皮层区域中[14C]葡萄糖合成谷氨酸的比例时,得到了相反的结果,即额叶皮层中谷氨酸的释放量在统计学上高于枕叶皮层。

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