Abstract
The influence of the herbicide 2,4-Dichlorophenoxyacetic acid on the growth of cultured chicken muscle cells was studied by observing morphological changes and by cytophotometric measurement of the DNA content of individual cells. DNA-synthesis curves were derived from the distribution of DNA values. No marked effects were observed at a concentration of 60 μg per liter. In contrast, doses of 600 μg per liter caused a loss in cell differentiation consisting of a partial lack of polar orientation, diminished formation of fibers, and an increase of the N/C ratio. The nuclear matrix of desoxyribonucleoprotein (DNP) changed to a more heteropycnotic structure. Isolated cell necroses were found after 44 hrs of exposure. When compared to the controls, the relative DNA synthesis time was found to be prolonged 1.5-fold. The number of cells in the G2 phase decreased during 44 hrs of exposure. It is concluded from this results that the dosedependent growth inhibition of 2,4-D on vertebrate cells in culture takes effect during DNA synthesis. The morphological changes of the nuclear structure suggest that 2,4-D directly affects the template activity of the DNP matrix.
Zusammenfassung
Der Einfluß des Herbicides 2,4-Dichlorphenoxyessigsäure auf das Wachstum von Skelettmuskelzellen des Huhns in Zellkulturen wurde anhand morphologischer Veränderungen und durch cytophotometrische Bestimmung des DNS-Gehaltes von Zellkernen untersucht. Aus der Verteilung der DNS-Meßwerte wurden Synthesekurven abgeleitet. Bis zu Konzentrationen von 60 μg/ml waren keine deutlichen Effekte erkennbar. Dagegen verursachten Konzentrationen von 600 μg/ml einen Verlust der Zelldifferenzierung, bestehend in einem Fehlen der polaren Wachstumsrichtung der Zellen, verminderter Faserbildung und einem Ansteigen der Kern-Plasma-Relation. Damit war eine Heterochromatisierung des Desoxyribonucleoproteins in den Zellkernen verbunden. Nach 44stündiger Einwirkung traten vereinzelte Zellnekrosen auf. Die relative DNS-Synthesedauer war gegenüber den Kontrollen auf das 1,5fache verlängert. Die Anzahl der Zellen in der G2-Phase war nach 44 Std Einwirkungsdauer vermindert. Aus den Befunden wird auf eine dosisabhängige Wachstumshemmung von Vertebraten-Zellen durch 2,4-D geschlossen, die während der DNS-Synthese wirksam ist. Die morphologischen Veränderungen der Kernmatrix legen die Vermutung nahe, daß durch 2,4-D die Aktivität des Chromatins direkt beeinflußt wird.
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Dedicated to Professor Wilhelm Doerr on his 60th birthday.
Supported by the Deutsche Forschungsgemeinschaft.
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Haag, D., Goerttler, K. & Preiss, D. The influence of non-cytotoxic concentrations of the herbicide 2,4-dichlorophenoxyacetic acid on the DNA synthesis in cultured vertebrate cells. Arch. Toxicol. 33, 91–102 (1975). https://doi.org/10.1007/BF00353234
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DOI: https://doi.org/10.1007/BF00353234