Biochemical analysis of cell architectonics in normal and diseased brain - Quantitative analysis at the single cell level -

取非神经系统疾病患者和神经系统疾病患者的中枢神经系统组织制作冻干切片。分离单个神经元，取小脑层和尾状核的显微标本。测定胆碱乙酰转移酶(ChAT)、谷氨酸脱羧酶(GAD)活性和γ-氨基丁酸(GABA)浓度。检测视网膜中GAD和GABA的含量。来自视网膜退化的小鼠。(1)ALS运动神经元因不明原因变性，至晚期萎缩消失。非进展期病例脊髓切片前角仍可见大量大的运动神经元胞体，进展期病例可见少量萎缩、脆弱的胞体。其余大细胞体的ChAT活性远低于对照细胞体。在萎缩的细胞体中，随着病理…的进展，其活性升高更符合逻辑的过程。推测萎缩的细胞体中产生了更多的ChAT，这些细胞体向出芽的许多轴突终末供应ChAT，以补偿细胞变性造成的肌肉神经支配的损失。(2)脊髓运动神经元是一种典型的胆碱能神经元，在动物体内含有GAD，GABA被认为是一种维持细胞功能和与肌肉突触接触的营养因子。对照Numan运动神经元中GAD活性很低。ALS运动神经元GAD活性无明显变化。(3)人小脑浦肯野细胞的GAD活性明显低于兔和大鼠。与动物标本相比，人小脑尾状核、分子层和颗粒层的活性也较低。这些结果表明，人神经元的递质合成活性普遍低于动物神经元。(4)变性视网膜的GABA能无长突细胞的GAD活性和Gabe浓度均显著高于正常视网膜。GABA被认为在机体退变过程中起营养因子的作用。较少

Freeze-dried sections of CNS tissues were prepared from control patients with non-neurological disease and patients with neurological diseases. Single neurons, as shown below, were isolated and microsamples of cerebellar layers and caudate nucleus were dissected out from these sections. Choline acetyltransferase (ChAT) and glutamate decarboxylase (GAD) activities and <gamma>-aminobutyric acid (GABA) concentration were assayed in these samples. GAD and GABA were analyzed in the retinas. from retinal-degeneration mice. (1) ALS motor neurons degenerate due to unknown etiology and atrophy to disappear at the far advanced stage. Many large cell bodies of motor neurons remained in the anterior horn of spinal cord section from non-advenced cases but a few atrophied and fragile cell bodies were detected with advanced cases. ChAT activity was far lower in the remaining large cell bodies than in control cell bodies. The activity became higher in the atrophied cell bodies with progress of patholo … More gical process. It was speculated that ChAT was produced much more in the atrophied cell bodies which supply ChAT to the many axon terminals sprouting to compensate innervation loss o fmuscle caused by cell degeneration. (2) Spinal motor neurons, a typical cholinergic neuron, contained GAD in animals and GABA was thought to be a kind of trophic factor to maintain the cell function and synaptic contact with muscle. Control numan motor neurons contain very low GAD activity. No change was observable in GAD activity of ALS motor neurons. (3) Human Purkinje cells from cerebellum had GAD activity much lower than those from rabbit and rat. Compared with animal samples, the activities were also lower in human caudate nucleus and in the molecular and granular layers from human cerebellum. These suggest that transmittersynthesyzing activity of human neuron is generally lower than that of animal neurons. (4) GABAergic amacrine cells of degenerating retinas contained markedly high levels of GAD activity and GABE concentration, compared with those in normal retina. GABA was thought to play a role as a trophic factor in response to degeneration process. Less