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Bindungsort und Bindungsmodus von Aldehydfuchsin in den B-Zellen des Inselapparates

Histochemische und elektronenmikroskopische Untersuchungen

The site and nature of aldehyde fuchsin staining of pancreatic beta granules

A study by histochemistry and electron microscopy

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Summary

In a combined study by histochemistry and electron microscopy some characteristics of aldehyde fuchsin were studied with special reference to its ability to stain pancreatic beta granules without previous oxydation.

  1. 1.

    Blocking of aldehyde fuchsin staining is possible by vigorous methylation and by acetylation. It is also possible to block the reaction by treating sections with different alcohols at higher temperatures in presence of hydrochloric acid. It is concluded, that the chemical group, which reacts in the tissue and which binds aldehyde fuchsin, is an enol. The blocking reactions are interpreted as a formation of enol ethers.

  2. 2.

    In the electron microscope the site of dye binding of aldehyde fuchsin could be detected by a new developed aldehyde fuchsin-ferric ferricyanide method. It is shown that in unoxydized sections the membranous sac of the beta granule and not the granule itself is stained by aldehyde fuchsin. Thus it seems to be very likely, that it is not insulin, which is stained by aldehyde fuchsin.

Zusammenfassung

Histochemisch und elektronenmikroskopisch wurde die Eigenschaft des Aldehydfuchsins (Gomori, 1950), ohne Voroxydation β-Granula in endokrinen Zellen des Pankreas anzufärben, untersucht.

  1. 1.

    Die Anfärbung von β-Granula mit Aldehydfuchsin wird blockiert durch vorhergehende Methylierung des Gewebes mit Methanol/Thionylchlorid oder durch Acetylierung mit Acetanhydrid/Pyridin. Außerdem wird die Anfärbbarkeit aufgehoben durch Behandlung mit verschiedenen Alkoholen in Wärme in Anwesenheit von Salzsäure. Es wird geschlossen, daß die im Gewebe reagierende und das Aldehydfuchsin bindende Gruppe ein Enol ist. Die Blockierungsreaktionen mit den Alkoholen werden dabei als Bildung von Enoläthern aufgefaßt.

  2. 2.

    Aldehydfuchsin vermag im Reagenzglas und im histologischen Schnitt Ferriferricyanidreagens rasch zu Berliner Grün zu reduzieren.

  3. 3.

    Der Ort der Farbstoffbindung des Aldehydfuchsin wird im Elektronenmikroskop durch die Aldehydfuchsin-Ferriferricyanid-Methode sichtbar gemacht. Es zeigt sich, daß im nichtoxydierten Schnitt das Aldehydfuchsin die Membran der β-Granula und nicht die Granula selbst anfärbt. Dabei wird sehr wahrscheinlich nicht Insulin mit Aldehydfuchsin nachgewiesen.

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  1. Abteilung mit Lehrstuhl für submikroskopische Pathologie und Neuropathologie der Universität Tübingen, Bundesrepublik Deutschland

    Hans v. Denffer

  2. Augenklinik r. d. Isarder Technischen Universität München, Ismaningerstraße 22, D-8000, München 80, Bundesrepublik Deutschland

    Hans v. Denffer

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Denffer, H.v. Bindungsort und Bindungsmodus von Aldehydfuchsin in den B-Zellen des Inselapparates. Histochemie 36, 97–113 (1973). https://doi.org/10.1007/BF00304386

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