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鸟嘌呤酶的组织化学显示:在人类中枢神经系统中的观察

The histochemical demonstration of guanase: observations in the human central nervous system.

作者信息

Norstrand I, Libbin R, Druck P, Enker S

出版信息

Acta Histochem. 1979;64(2):187-93. doi: 10.1016/S0065-1281(79)80072-0.

Abstract
  1. A method is described for the histochemical demonstration of the purine catabolizing enzyme guanase, employing glutaraldehyde fixation and Nitro blue tetrazolium (NBT). Parallel biochemical studies confirm that enzyme activity is not significantly inhibited by exposure to glutaraldehyde. 2. By this procedure guanase activity has been visualized in neurons and glial elements of the human central nervous system (CNS). 3. Controls consisted of direct incubation of cryostat sections with a specific inhibitor of guanase (5-amino-4-imidazole carboxamide) and omission successively of the substrate guanine, of xanthine oxidase and of NBT. Enzyme activity was completely inhibited by the above procedures, and by boiling of tissues for 10 min prior to fixation. 4. Levels of enzyme activity in spinal cord and brain were assessed by a subjective scoring method, and showed close comparability with biochemical assay data in brainstem and cerebral hemispheres; whereas a low correlation for enzyme activity was observed in spinal cord and cerebellum. Differences between biochemical and histochemical assessments of CNS guanase activity are discussed.
摘要
  1. 本文描述了一种用于嘌呤分解代谢酶鸟嘌呤酶组织化学显示的方法,该方法采用戊二醛固定和硝基蓝四唑(NBT)。平行的生化研究证实,暴露于戊二醛不会显著抑制酶活性。2. 通过该程序,已在人类中枢神经系统(CNS)的神经元和神经胶质成分中观察到鸟嘌呤酶活性。3. 对照包括用鸟嘌呤酶的特异性抑制剂(5-氨基-4-咪唑甲酰胺)直接孵育冰冻切片,以及依次省略底物鸟嘌呤、黄嘌呤氧化酶和NBT。上述程序以及固定前将组织煮沸10分钟可完全抑制酶活性。4. 通过主观评分法评估脊髓和脑中的酶活性水平,结果显示与脑干和大脑半球的生化测定数据具有高度可比性;而在脊髓和小脑中观察到酶活性的相关性较低。文中讨论了中枢神经系统鸟嘌呤酶活性的生化和组织化学评估之间的差异。

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