Summary
The splitting of 6-Br-2-naphthyl-, α-naphthyl-, and 4-Cl-5-Br-3-indolyl-glycosides which proved useful for the assessment of cytological localization of intestinal enzymes in previous studies was investigated using isolated human and rat intestinal disaccharidases as a source of enzyme activities.
Previous findings based on histochemical studies were confirmed and extended. 6-Br-2naphthylα-D-glucoside is cleaved by glucoamylase and sucrase-isomaltase. The participatio of trehalase in splitting of this substrate is very low and can be neglected. The mentioned α-glucosidases are responsible for the brush border staining of enterocytes with this substrate when unfixed cold microtome sections are used. Even when a differential heat inactivation of sucrase-isomaltase and of glucoamylase occurs during paraffin embedding (so that the staining in paraffin sections is due mostly to glucoamylase) the use of natural substrates is desirable for a more precise assessment of sucrase-isomaltase activity (but without the possibility of a correct localization).
4-Cl-5-Br-3-indolyl-β-D-fucoside is the substrate of choice for the demonstration of lactase. Even when this substrate is split also by “hetero-β-galactosidase” and by acid (lysosomal) β-galactosidase these activities do not disturb the histochemical demonstration of lactase. If however some doubts arise, the inhibition with p-Cl-mercuribenzoate (2 · 10−4 M) is to be emloyed (lactase activity is not inhibited). Due to a low Km and a high Vmax of indolyl-fucoside and due to its extreme stability in solution (which enables to use the substrate solution repeatidly) this substrate is suitable in routine practice even though it is expensive.
α-naphthyl- and 4-Cl-5-Br-3-indolyl-β-D-glucosides are split by lactase and β-glucosidase. Due to the fact that the mutual delineation of these activities is not easy and that Km an Vmax for lactase are not so favourable as in the case of fucoside these substrates are not recommended for the assessment of lactase.
6-Br-2-naphthyl-β-D-glucoside is the substrate of choice for the histochemical studies concerned with “hetero-β-galactosidase” and 4-Cl-5-Br-3-indolyl-β-D-galactoside for acid β-galactosidase.
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Lojda, Z., Slabý, J., Kraml, J. et al. Synthetic substrates in the histochemical demonstration of intestinal disaccharidases. Histochemie 34, 361–369 (1973). https://doi.org/10.1007/BF00306308
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DOI: https://doi.org/10.1007/BF00306308