HISTOCHEMICAL DEMONSTRATION OF N-ACETYL-BETA-GLUCOSAMINIDASE EMPLOYING NAPHTHOL AS-BI N-ACETYL-BETA-GLUCOSAMINIDE AS SUBSTRATE
HISTOCHEMICAL DEMONSTRATION OF N-ACETYL-BETA-GLUCOSAMINIDASE EMPLOYING NAPHTHOL AS-BI N-ACETYL-BETA-GLUCOSAMINIDE AS SUBSTRATE
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DOI：
10.1177/13.5.355
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发表时间：
1965-01-01
影响因子：
3.2
通讯作者：
HAYASHI, M
中科院分区：
文献类型：
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作者：
HAYASHI, M
萘酚AS-BI(7-溴-3-羟基-2-萘-邻-茴香胺)的N-乙酰基-β-氨基葡萄糖胺是通过使茴香胺与乙酰氯葡萄糖胺在碱性丙酮水溶液中反应获得的。用甲醇氨除去O-乙酰基并从甲醇水溶液中重结晶后，该化合物可以用作N-乙酰基-β-氨基葡萄糖苷酶的底物。在副玫瑰苯胺六氮盐或快速石榴石 GBC 存在下，在 pH 5.2 下，将甲醛-钙固定的冷冻切片与 0.5 m[图像] 底物一起孵育。染色在细胞质中主要以离散颗粒的形式出现在假定的酶活性位点，并且在 N-乙酰氨基葡萄糖内酯存在下被抑制。在大鼠肾脏中，在近曲小管的近端部分观察到许多被酶染色的球形颗粒。在肝脏中，染色被认为是位于实质肝细胞的细管周细胞质中的小颗粒。这些组织中的一般细胞学定位与其他溶酶体酶的定位非常相似。简要讨论了该技术的局限性。
The N-acetyl-[beta]-glucosaminide of naphthol AS-BI (7-bromo-3-hydroxy-2-naphth-o-anisidide) was obtained by reacting the anisidide with acetochlorogluco-samine in aqueous alkaline acetone. After removal of O-acetyl groups with methanolic ammonia and recrystallization from aqueous methanol, the compound may be used as a substrate for N-acetyl-[beta]-glucosaminidase. Formol-calcium fixed frozen sections were incubated with 0.5 m[image] substrate in the presence of hexazonium pararosanilin or fast garnet GBC at pH 5.2. Staining appeared in the cytoplasm mostly as discrete granules at the presumed site of enzyme activity and was inhibited in the presence of N-acetylglucosaminolactone. In the rat kidney a number of spherical granules which stained for the enzyme were observed in the proximal portion of the proximal convoluted tubules. In the liver staining was recognized as small granules localized at the pericanalicular cytoplasm of parenchymal hepatic cells. The general cytological localization in these tissues was quite similar to that of other lysosomal enzymes. The limitations of the technique were briefly discussed.