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DNA DSBs repair kinetics in neurons and astrocytes of primary hippocampal cell culture after irradiation with Co60 g-rays and proton
Abstract
The induction and repair of DNA double-strand breaks (DNA DSB) were analyzed in primary hippocampal cell culture under the action of Co60 g-rays or protons at 3 Gy dose. For investigation of DNA DSBs formation, we established two cell cultures: neuronal cell culture, created with using of antimitotic agent 1-β-D-arabinofuranosil cytosine (AraC), and primary hippocampal cell culture without influence of Ara-C includes both neuronal and glial elements. The study of DNA DSB formation and repair in neuron and primary cell culture was conducted using DNA repair protein markers – γH2AX and 53BP1.
It was established that the gH2AX/53BP1 foci quantity reached the maximum 1h after both types of radiation and decrease in 24 h post- irradiation. However, 24h post-irradiation the radiation-induced foci (RIF) level remained significantly different to non-irradiated samples. In case of proton irradiation, a higher number of RIF was observed 24 h after exposure compare to γ-irradiation.
The study of the formation and elimination kinetics of γH2AX foci in primary hippocampal cell culture showed the maximum of foci number 1 h after exposure to γ-rays. There was a delay in foci elimination, followed by an increase in the number of RIF 4 h after proton irradiation. The structure of RIF clusters in astrocytes becomes more complex in comparison with neuronal cells after exposure to both types of irradiation throughout the entire post-radiation period.
