Acta Vet. Brno 2011, 80: 113-118

Phosphorylation of histone H2AX as an indicator of received dose of gamma radiation after whole-body irradiation of rats

Radim Havelek1, Martina Řezáčová1, Zuzana Šinkorová2, Lenka Zárybnická2, Aleš Tichý1,2, Jiřina Vávrová2

1Department of Medical Biochemistry, Faculty of Medicine in Hradec Králové, Charles University in Prague, Czech Republic
2Department of Radiobiology, University of Defence, Faculty of Military Health Sciences, Hradec Králové, Czech Republic

The aim of our study was to determine whether phosphorylation of histone H2AX can be used as an indicator of received dose of gamma radiation after whole-body irradiation of rats. Wistar rats were irradiated by 1-10 Gy of gamma radiation by 60Co source. Value LD50/60 was 7.37 (4.68-8.05) Gy. Histone H2AX is phosphorylated by ATM kinase on serine 139 (γH2AX) quickly after the irradiation. It forms microscopically visible foci in the site of double strand breaks of DNA. Flow-cytometric method was used for quantitative detection. This study is the first one that evaluated dose-dependency of H2AX phosphorylation in peripheral lymphocytes of rats irradiated by whole-body dose 1-10 Gy. Our data show a dose-dependent increase in γH2AX in rat peripheral blood lymphocytes 1 h after whole-body irradiation by the dose of 1-10 Gy. We proved that phosphorylation of histone H2AX is a prompt and reliable indicator of the received radiation dose suitable for rapid measurement before the number of lymphocytes in peripheral blood starts to decrease. It can be used already 1 h after the irradiation for an estimation of the received dose of radiation. Blood samples can be stored in 4 °C for 23 h without significantly affecting the result.


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