Summary
Glucagon-storing cells were detected in the pancreatic islets of horse, guinea pig, rabbit, rat, dog and man by means of indirect immuno-histochemical procedures using fluorescein or peroxidase as markers of antiglucagon sera. By subsequently staining horse islets with selective staining techniques for A, B or D cells, it was directly ascertained that only A cells reacted to antiglucagon sera. A comparison of methods known to stain a granules with immuno-histochemical methods, as well asin vitro experiments on natural and synthetic glucagon, showed that the xanthydrol method, and perhaps also theo-phthalaldehyde method, should be regarded as specific histochemical tests for glucagon whereas the HCl-basic dye technique and phosphotungstic haematein should be considered as unspecific glucagon-staining methods,and Grimelius' silver as a method likely to be unrelated to glucagon. In ultrastructural investigations on the human pancreas, the core of the a granule heavily reacted to phosphotungstic acid and peroxidase-marked antiglucagon antibodies, while being unreactive to Grimelius' silver; conversely, the peripheral halo of theα granule heavily reacted to Grimelius' silver, while being poorly reactive or unreactive to phosphotungstic acid and anti-glucagon antibodies. Thus, both light and electron microscopy findings point to a structural and chemical hetereogeneity of theα granule.
Résumé
Des cellules stockant du glucagon ont été détectées dans les îlots pancréatiques du cheval, du cobaye, du lapin, du rat, du chien et de l'homme au moyen de procédés immuno-histochimiques indirects utilisant la fluorescéine ou la peroxydase comme marqueurs des sérums anti-glucagon. En colorant les îlots des chevaux avec des techniques de coloration sélectives pour les cellules A, B ou D, il a été directement prouvé que seules les cellules A réagissent aux sérums anti-glucagon. Une comparaison entre les méthodes connues pour colorer les granulesα et les méthodes immuno-histochimiques, ainsi que les expériencesin vitro sur le glucagon naturel et synthétique, ont montré que la méthode au xanthydrol et peut-être également la méthode à l'o-phtalaldéhyde, doivent être considérées comme des tests histochimiques spécifiques pour le glucagon; par contre la technique à l'HCl-colorant basique et l'hématéine phosphotungstique doivent être considérées comme des méthodes de coloration non spécifiques pour le glucagon, et la méthode à l'argent de Grimelius probablement comme ne concernant pas le glucagon. Au cours d'investigations ultrastructurales sur le pancréas humain, le centre des granulesα réagissait fortement à l'acide phosphotungstique et aux anticorps anti-glucagon marqués à la peroxydase, alors qu'il ne réagissait pas à l'argent de Grimelius; inversement le halo périphérique des granulesα réagissait fortement à l'argent de Grimelius, et réagissait peu ou pas à l'acide phosphotungstique et aux anticorps anti-glucagon. Ainsi, les résultats à la fois de microscopie optique et électronique indiquent une hétérogénéité structurale et chimique des granulesα.
Zusammenfassung
Die Glucagon enthaltenden Zellen werden in der Bauchspeicheldrüse von Pferd, Meerschweinchen, Kaninchen, Ratte, Hund und Mensch durch immunhistochemische, indirekte Techniken mit Fluorescein-oder Peroxidase-Markierung von Anti-GlucagonAntikörpern lokalisiert. Durch aufeinanderfolgende Färbungen der Langerhansschen Inseln von Pferden mit spezifischen für die A, B und D Zellen histologischen Methoden, wurde bewiesen, daß nur die A Zellen mit AntiGlucagon-Antikörpern reagierten. Eine Gegenüberstellung der für die Färbung desα-Granulums bekannten Methoden, mit den immunhistochemischen Methoden, und den alsin vitro Untersuchungen über synthetisches und natürliches Glucagon, haben bewiesen, daß die Xanthydrol-Methode, und vielleicht auch dieo-Phtalaldehyd Methode, für das Glucagon spezifische Methoden darstellen, während die HCl-basischen Farbstoffe und die Phosphorwolframsäure-Hämatin Methoden, als unspezifische Färbemethoden für das Glucagon angesehen werden müssen; die Silbermethode nach Grimelius scheint nicht zum Glucagon in Beziehung zu stehen. In ultrastrukturellen Untersuchungen über die menschliche Bauchspeicheldrüse, reagierte das Zentrum (core) des Granulums stark auf die Phosphorwolframsäure und auf die mit Peroxidase markierten Anti-Glucagon-Antikörper, dagegen reagierte es nicht mit der Silbermethode nach Grimelius. Andererseits reagierte der periphere Hof desα Granulums wenig, oder überhaupt nicht mit der Phosphorwolframsäure und den Anti-Glucagon-Antikörpern. Daher zeigen die Ergebnisse von Licht-und Elektronenmikroskopie eine chemische und strukturelle Ungleichartigkeit desα-Granulums.
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Bussolati, G., Capella, C., Vassallo, G. et al. Histochemical and ultrastructural studies on pancreatic A cells. Evidence for glucagon and non-glucagon components of the α granule. Diabetologia 7, 181–188 (1971). https://doi.org/10.1007/BF01212551
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DOI: https://doi.org/10.1007/BF01212551