Summary
Beta cell replication was studied in normal (C 57 BL/Ks) and diabetic mutant (C 57 BL/Ks-db/db) mice following thymidine-3H administration. The specific activity of DNA of isolated islets (DPM/μg islet DNA) was used as an index of proliferative activity and correlated with labeling determined by radioautography. Although thymidine-3H incorporation in islets of prehyperglycemic 5 to 6 week old mutants was limited, it was significantly greater than that in normal mice. With the elevation of blood glucose values, incorporation rose sharply, reaching a maximum level above 130 mg glucose/100 ml blood. Sustained, severe hyperglycemia subsequently correlated with a decline in islet DNA synthesis. Food restriction early in the syndrome reduced hyperglycemia and resulted in low incorporation of label. Animals refed ad lib for periods of 1, 2, or 3 weeks showed significant increases in labeling, with maximal values after 1 week of refeeding. Electron microscopic radioautographs of the islets revealed labeled beta cells but no labeled alpha cells, suggesting that proliferative activity is predominantly restricted to the beta cell population.
Résumé
La division des cellulesβ a été étudiée chez la souris normale (C 57 BL/Ks) et chez son mutant diabétique (C 57 BL/Ks-db/db) après administration de3H-thymidine. L'activité spécifique de l' ADN des îlots isolés (DPM/μg ADN) sert d'index de l'activité de prolifération et est corrélée avec le marquage décélé par autoradiographie. Bien que l'incorporation de3H- thymidine dans les îlots de sourisdb/db préhyperglycémiques âgées de 5 à 6 semaines soit assez faible, elle est cependant significativement plus élevée que chez la souris normale. L'augmentation des taux de glucose sanguin s'accompagne d'un accroissement très net de l'incorporation, qui est maximale pour les glycémies supérieures à 130 mg glucose %. Une hyperglycémie sévère et prolongée s'accompagne à la suite d'une diminution de la synthèse d'ADN dans les îlots. Une diminution de l'apport alimentaire au début de la manifestation du syndrome décroît l'hyperglycémie concomitante avec une incorporation faible de la substance marquée. Lorsque ces animaux peuvent de nouveau s'alimenter à volonté pendant 1, 2 ou 3 semaines, on constante une augmentation significative du marquage atteignant un taux maximum après une semaine de réalimentation. — L'examen autoradiographique combiné à la microscopie électronique des îlots démontre la présence de radioactivité dans les cellulesβ, mais pas dans les cellulesα. Ce résultat suggère que l'activité de prolifération est essentiellement confinée aux cellulesβ.
Zusammenfassung
Mittels Messung der spezifischen Aktivität der DNA isolierter Langerhans'scher Inseln und radioautographischer Lokalisation der Radioaktivität nach Injektion von Thymidin-3H wurde die Vermehrung der B-Zellen im Pankreas normaler und hereditär diabetischer (db/db) Mäuse untersucht. Die Inkorporation von Radioaktivität war bei 5–6 Wochen altendb/db Tieren signifikant höher als bei den normalen Kontrolltieren. Mit Ansteigen der Blutzuckerkonzentration nahm die Inkorporation zu und erreichte bei Blutzuckerkonzentrationen über 130 mg/100 ml ihr Maximum. Anhaltend schwere Hyperglykämie war von einem progressiven Abfall der Inkorporation von Radioaktivität in die Insel-DNA begleitet. Kalorienrestriktion in einem Frühstadium des Syndroms verminderte sowohl die Hyperglykämie als auch die Inkorporation von Radioaktivität. Nach Normalisierung der Nahrungsaufnahme zeigte sich eine deutliche Erhöhung der Inkorporation, die nach einer Woche ihr Maximum erreichte. Elektronenmikroskopische Autoradiographie ergab, daß die Radioaktivität ausschließlich in den B-Zellen lokalisiert war. Markierte A-Zellen wurden nicht beobachtet. Dies weist darauf hin, daß sich die proliferative Aktivität praktisch ausschließ-lich auf die B-Zellen beschränkt.
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Supported in part by USPHS Grants AM-12538, AM-09584, and AM-05077.
USPHS Special Postdoctoral Fellowship Awardee, Grant F3-AM-36335.
USPHS Research Career Development Awardee, Grant K4-AM-7394.
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Chick, W.L., Like, A.A. Studies in the diabetic mutant mouse: III. Physiological factors associated with alterations in beta cell proliferation. Diabetologia 6, 243–251 (1970). https://doi.org/10.1007/BF01212233
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DOI: https://doi.org/10.1007/BF01212233