Summary
With the electron microscopic demonstration of esterases the question arises, how far the natural condition of tissue esterase is qualitatively or quantitatively changed by the preceeding fixation. In three organs of the mouse which are rich in esterase (liver, kidney and the small intestine), the activity of esterase before and after fixation with glutardialdehyde (GA) and formaldehyde (FA) is determined against four substrates. These substrates are used, as well, for electron microscopic demonstration of the enzyme. Thereby, the fixation conditions usually applied in electron microscopic techniques were exactly copied.
The loss of esterase activity in liver and kidney amounts to 70% after GA-fixation and 50% after FA-fixation, independent of the substrates used. That means that only quantitative but no qualitative changes of esterase composition could be found. Therefore there is no reason to assume that the main esterase-components in natural tissues are not demonstrated in the electron microscopical picture. Because of the considerable esterase content of mucus, the small intestine gives different quantitative results.
The multiple conditions which are responsible for the degree of esterase inactivation by fixing agents are discussed.
Zusammenfassung
Bei der Darstellung von Esterasen im Elektronenmikroskop taucht die Frage auf, in welcher Weise der native Zustand der Gewebeesterase durch die vorausgehende Fixation qualitativ und quantitativ verändert wird. In den esterasereichen Organen der Maus (Leber, Niere und Dünndarm) wird die Esteraseaktivität gegenüber vier auch in der Elektronenmikroskopie eingesetzten Substraten vor und nach Fixation mit Glutardialdehyd (GA) bzw. Formalin (FA) bestimmt. Dabei werden die in der Elektronenmikroskopie üblichen Fixationsbedingungen genau kopiert.
Der Aktivitätsverlust der Esterase beträgt in Leber und Niere bei allen Substraten 70% nach GA-Fixation und 50% nach FA-Fixation, d.h., es sind zwar quantitative, aber keine qualitativen Veränderungen der Esterasezusammensetzung nachzuweisen. Es ergibt sich damit kein Anhaltspunkt dafür, daß die Hauptkomponenten der im nativen Gewebe vorkommenden Esterase im elektronenmikroskopischen Bild nicht erfaßt werden. Wegen des erheblichen Esterasegehaltes im Schleim werden im Darm andere quantitative Verhältnisse gefunden.
Die vielfältigen Bedingungen, von denen das Ausmaß der Esteraseinaktivierung durch Fixationsmittel abhängt, werden diskutiert.
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Böcking, A., Großarth, C. & Deimling, O.v. Esterase. Histochemie 37, 265–273 (1973). https://doi.org/10.1007/BF00304187
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DOI: https://doi.org/10.1007/BF00304187