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Zur Wirkung und Lokalisation von H3-actinomycin D in Eifollikeln von Musca domestica nach in vivo-Applikation

Effects and localization of H3-actinomycin D in the egg follicle of Musca domestica after in vivo application

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Summary

Actinomycin effects on cell structure and its intracellular localization were studied in growing egg follicles of Musca domestica. 16 females at different stages of oogenesis were injected with 0,4 μg of H3-actinomycin. After incubation times of 30 min, 24 hrs, or 48 hrs, the flies were killed and the ovaries prepared for autoradiography. Control slides were stained with Azur B after RNase treatment to demonstrate the DNA distribution. In the short time experiments, actinomycin was always found in connection with nuclear DNA. The labeling pattern corresponded to the DNA content of the several cell types and therefore differed in the diploid ovariole envelop cells, the polyploid follicular cells, and the highly polyploid nurse cells. In the germinal vesicle only the karyosphere was labeled. A rather weak labeling of the ooplasm in autoradiographs exposed for 6 months is probably due to H3-actinomycin bound by the mitochondrial DNA. No radioactivity could be detected in the yolk platelets. The DNA-actinomycin-complex was found to have a high stability. During 48 hrs of incubation no dislocation or loss of radioactivity could be observed.

A long time treatment with actinomycin releases the degeneration of the trophic chamber already at stage 3 and 4 which normally occurs at stage 5 of follicle development. The giant nurse cell nuclei become pycnotic. The oocyte nucleus is often discharged into the central ooplasm. The structure of its karyosphere and the endobody appear to be unchanged even by long time treatment with actinomycin, after which the cellular contact between the cells of the follicle epithelium is broken down.

Zusammenfassung

An 16 Stubenfliegen- ♀♀ der Oogenese-Stadien 2–6 wurden nach Inkubationszeiten von 30 min bis 48 h Wirkung und Verbleib von H3-Actinomycin untersucht. Die Markierungsverteilung im Autoradiogramm entspricht dem mit Azur B-Färbung nach RNase-Vorbehandlung dargestellten DNS-Gehalt der Kerne der verschiedenen Zellarten im Ovar. Die Riesenkerne der hochpolyploiden Nährzellen binden Actinomycin am stärksten. Bei den Oocytenkernen liegen Silberkörner nur über dem Bereich der Karyosphäre. Eine längere Einwirkung von Actinomycin verursacht degenerative Veränderungen der Kern-und Gewebestruktur des Ovars. Der DNS-Actinomycin-Komplex bleibt mindestens 48 h erhalten, es finden sich keine Anzeichen für eine Verlagerung oder einen Abbau des Antibioticums während dieser Zeit. Eine Bindung in den Dotterschollen wurde nicht festgestellt. Eine nach langen Expositionszeiten beobachtete schwache autoradiographische Markierung des Cytoplasmas älterer Oocyten wird vermutlich durch eine Actinomycin-Anlagerung an die DNS der Mitochondrien verursacht.

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  1. Zoologisches Institut der Universität Münster, Germany

    Wolf Engels

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Engels, W. Zur Wirkung und Lokalisation von H3-actinomycin D in Eifollikeln von Musca domestica nach in vivo-Applikation. Histochemie 19, 224–234 (1969). https://doi.org/10.1007/BF00305285

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