NACHLAS M M, WALKER D G, SELIGMAN A M
J Biophys Biochem Cytol. 1958 Jan 25;4(1):29-38. doi: 10.1083/jcb.4.1.29.
The present investigation concerning the histochemical demonstration of DPN diaphorase follows the development of a new reagent, Nitro-BT, which has already been used successfully for the cytochemical localization of the succinic dehydrogenase system. The most consistently favorable results were obtained with the lactate-lactic dehydrogenase system buffered at pH 7.4. Using sections of rat kidney and stomach, it was found that the intensity of stain was optimal after 15 minutes incubation at 37 degrees C., conducted aerobically. By appropriate variations in the substrate mixture it was possible to selectively demonstrate the histochemical distribution of certain DPN-linked dehydrogenases in addition to DPN diaphorase. This was made possible by the special distribution of some of these dehydrogenases which distinguished them from one another. Of the dehydrogenases studied the distribution pattern of beta-hydroxybutyric dehydrogenase was the most singular. In the gastric mucosa beta-hydroxybutyric dehydrogenase was restricted to the cells of the mucous lining epithelium and the gland necks; and in the kidney the enzyme was limited to the cells of the proximal convoluted tubule and thick limbs of Henle's loop. In contrast, lactic dehydrogenase like DPN diaphorase was demonstrable in almost all cytologic elements of both the stomach and the kidney.
本次关于二磷酸吡啶核苷酸（DPN）黄递酶组织化学显示的研究，是在一种新试剂硝基蓝四氮唑（Nitro - BT）问世后开展的，该试剂已成功用于琥珀酸脱氢酶系统的细胞化学定位。在pH 7.4缓冲的乳酸 - 乳酸脱氢酶系统中获得了最为一致的良好结果。使用大鼠肾脏和胃的切片，发现在37℃有氧条件下孵育15分钟后染色强度最佳。通过适当改变底物混合物，除了DPN黄递酶外，还能够选择性地显示某些与DPN相关的脱氢酶的组织化学分布。这是因为这些脱氢酶中的一些具有特殊分布，从而彼此区分开来。在所研究的脱氢酶中，β - 羟丁酸脱氢酶的分布模式最为独特。在胃黏膜中，β - 羟丁酸脱氢酶局限于黏液衬里上皮细胞和腺颈部细胞；而在肾脏中，该酶仅限于近端曲管和亨氏袢粗段的细胞。相比之下，乳酸脱氢酶与DPN黄递酶一样，在胃和肾脏的几乎所有细胞成分中都可显示。
