Contestabile A, Andersen H
Histochemistry. 1978 Jun 9;56(2):117-32. doi: 10.1007/BF00508438.
In a detailed study focused on the methodological problems in dehydrogenase histochemistry [e.g., fixation, diffusion of enzymes and of reduced inermediates, conversion of NADPH and NADP to NADH and NAD, respectively, penetration of tetrazolium salt and formazan substantivity, 'nothing dehydrogenase' reaction, use of exogenous CoQ10 and of flavoprotein substitute (PMS)], the distribution and activity of succinate dehydrogenase, NAD(P)H-tetrazolium reductase, glucose-6-phosphate dehydrogenase, lactate dehydrogenase (H and M types), and of L-glutamate dehydrogenase (E.C.1.4.1.2 and E.C.1.4.1.3) have been investigated in the rat cerebellum. It was evident from the study that reliable results could only be obtained if all the aforementioned factors had been considered. The image of actual concentration of SDH in the neuropil of the molecular layer could only be recorded by adding CoQ10, while other structures exhibited greater balance between SDH and endogenous mitochondrial CoQ. Contrary to previous studies, a reversed localization of the activity of G-6-PDH and LDH was noticed. The elements of molecular and Purkinje layers were rich in G-6-PDH, while the granular layer was nearly depleted. The actual level of LDH could only be recorded if NADH-tetrazolium reductase was bypassed with PMS. The H and M types of LDH coexisted in the three cortical layers, the H type being prevalent and the M type attaining its highest level in synaptic glomeruli followed by the structures of the molecular layer and the Purkinje cells. High activity of GDH was noticed in Bergmann glia followed by synaptic glomeruli, while most other structures showed weak to moderate activity. The two GDH types coexisted in all structures showing activity, except for Bergmann cells, which only showed presence of the E.C. 1.4.1.3 type. Furthermore, Bergmann glia was exceptional by showing no activity of SDH and LDH, but strong activity of G-6-PDH and NADPH-tetrazolium reductase. The granular cells were exceptional by showing weak or no activity of all enzymes in question.
在一项针对脱氢酶组织化学方法学问题的详细研究中[例如，固定、酶和还原中间体的扩散、NADPH和NADP分别转化为NADH和NAD、四氮唑盐的穿透和甲臜显色性、“无脱氢酶”反应、外源性辅酶Q10和黄素蛋白替代物（PMS）的使用]，研究了大鼠小脑琥珀酸脱氢酶、NAD(P)H-四氮唑还原酶、葡萄糖-6-磷酸脱氢酶、乳酸脱氢酶（H型和M型）以及L-谷氨酸脱氢酶（E.C.1.4.1.2和E.C.1.4.1.3）的分布和活性。研究表明，只有考虑到上述所有因素才能获得可靠的结果。只有添加辅酶Q10才能记录分子层神经毡中SDH的实际浓度图像，而其他结构在SDH和内源性线粒体辅酶Q之间表现出更大的平衡。与先前的研究相反，观察到G-6-PDH和LDH活性的定位相反。分子层和浦肯野层的成分富含G-6-PDH，而颗粒层几乎没有。只有用PMS绕过NADH-四氮唑还原酶才能记录LDH的实际水平。LDH的H型和M型共存于三个皮质层，H型占优势，M型在突触小球中达到最高水平，其次是分子层结构和浦肯野细胞。在伯格曼胶质细胞中观察到GDH的高活性，其次是突触小球，而大多数其他结构显示出弱至中度活性。两种GDH类型共存于所有有活性的结构中，除了伯格曼细胞，其仅显示E.C.1.4.1.3类型。此外，伯格曼胶质细胞的特殊之处在于它没有SDH和LDH的活性，但有很强的G-6-PDH和NADPH-四氮唑还原酶活性。颗粒细胞的特殊之处在于它对所有相关酶的活性都很弱或没有活性。
