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The histochemical demonstration of aminopeptidase with bromoindolyl leucinamide

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Summary

The indigogenic method for aminopeptidase of Pearson et al. (1963) was critically evaluated. The localization obtained with it is not correct due to diffusion artifacts. Ferricyanide cannot be used as an oxidation agent. Based on experiments with other oxidation agents (phenazonium methosulfate, nitro BT, tetranitro BT) a new method was devised.

The recommended incubation medium contains 0.9 mM L-N-(5-bromoindol-3-yl) leucinamide hydrobromide (chloride), 0.73 mM tetranitro BT, 0.27 mM phenazonium methosulfate and 0.67 M phosphate buffer pH 7.4. The enzyme activity is indicated by the deposition of tetranitro BT formazan.

Results with this method in rat kidney, jejunum, liver, lung, and submaxillary gland, in monkey kidney and jejunum, and in human jejunal biosies are almost identical with those obtained with L-leucyl-4-methoxy-β-naphthylamide applied in a simultaneous azocoupling procedure. The given principle of the demonstration of aminopeptidase activity with an indolylamine substrate deserves a further exploration in the demonstration of peptidases “in situ” both on optical as well as electronmicroscopical levels.

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Authors and Affiliations

  1. Research Laboratory of Angiology Faculty of General Medicine, Charles University, Prague, ČSSR

    Z. Lojda & E. Havránková

  2. 1st Department of Pathology, Faculty of General Medicine, Charles University, Prague, ČSSR

    Z. Lojda & E. Havránková

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  1. Z. Lojda
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  2. E. Havránková
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Dedicated to the 60th birthday of Prof. MUDr. J. Švejda, DrSc., Head of the Department of Pathology, Medical Faculty, Purkyně University, Brno.

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Lojda, Z., Havránková, E. The histochemical demonstration of aminopeptidase with bromoindolyl leucinamide. Histochemistry 43, 355–366 (1975). https://doi.org/10.1007/BF00490194

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  • DOI: https://doi.org/10.1007/BF00490194

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