HISTOCHEMICAL TRACING OF LYSOSOMAL-ENZYMES - IMPROVED PREPARATION TECHNIQUE FOR ACID-PHOSPHATASE, BETA-GLUCURONIDASE, ACID-BETA-GALACTOSIDASE, AND ARYLESTERASE IN RAT-LIVER

  • 1 January 1981
    • journal article
    • research article
    • Vol. 69  (1) , 1-+
Abstract
The efficiency of the membrane methods in histochemical lysosome tracing was studied and compared with the results of a preparation technique, employing fresh-frozen celloidin-coated sections; these methods were compared with results obtained with conventional formalin-sucrose-fixed livers. The lysosomes were traced by reactions for acid phosphatase .beta.-glucuronidase, arylesterase and acid-.beta.-galactosidase activity in rat livers systematically harvested at different time points of a 24 h period. Each preparation method revealed a circadian (day-time-dependent) variation of lysosomes with respect to their size, number and activity. The lysosomes traced with these 4 marker enzymes differed in their circadian phase, i.e., the time points of occurrence of their maximal and minimal activities during the 24 h period, thus indicating a heterogeneity of lysosomes. Optimal results with respect to the morphological appearance of lysosomes were obtained in fresh-frozen celloidin coated sections. This preparation method caused no enzyme inhibition or loss, thus delivering comparatively the strongest reactions in livers and in enzyme models. Day-time-dependent extralysosomal enzyme activities regularly occur in hepatocytes. Extralysosomal localizations are not a consequence of technically induced enzyme diffusion; they are best visualized in celloidin-coated sections; the membrane method produces less satisfactory results, and formalin-sucrose-fixed livers were least satisfactory.

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