Repair of DNA double-strand breaks is not modulated by low-dose gamma radiation in C57BL/6J mice.
In this study, we sought to determine whether low-dose ionizing radiation, previously shown to induce a systemic adaptive response in C57BL/6J mice, is capable of enhancing the rate of DNA double-strand break repair. Repair capacity was determined by measuring γ-H2AX levels in splenic and thymic lymphocytes, using flow cytometry, at different times after a challenge irradiation (2 Gy, (60)Co). Irradiation with low doses (20 and 100 mGy) was conducted in vivo, whereas the challenge dose was applied to primary cultures of splenocytes and thymocytes in vitro 24 h later. Obtained kinetics curves of formation and loss of γ-H2AX indicated that cells from low-dose irradiated mice did not express more efficient DNA double-strand break repair compared to controls. Immunoblot analysis of γ-H2AX and Phospho-Ser-1981 ATM confirmed that DNA damage signaling was not modulated by preliminary low-dose radiation. Mouse embryonic fibroblasts of C57BL genetic background failed to show clonogenic survival radioadaptive response or enhanced repair of DNA double-strand breaks as evaluated by immunofluorescence microscopy of γ-H2AX foci. Our results indicate that radiation adaptive responses at systemic levels, such as increases in the tumor latency times in aging mice, may not be mediated by modulated DNA repair, and that the genetic background may affect expression of a radioadaptive response.
a  Chalk River Laboratories, Atomic Energy of Canada Limited, Chalk River, Ontario, Canada;
Melinda S J
MS
Blimkie
Luke C W
LC
Fung
Eugenia S
ES
Petoukhov
Cyrielle
C
Girard
Dmitry
D
Klokov
D000818:Animals; D002478:Cells, Cultured; D003114:Colony-Forming Units Assay; D053903:DNA Breaks, Double-Stranded / Q000528:radiation effects*; D004249:DNA Damage; D004260:DNA Repair / Q000528:radiation effects*; D004307:Dose-Response Relationship, Radiation; D005260:Female; D005347:Fibroblasts / Q000528:radiation effects; D005720:Gamma Rays; D006657:Histones / Q000032:analysis; D059165:Hormesis; D016131:Lymphocyte Subsets / Q000378:metabolism / Q000528:radiation effects* / Q000648:ultrastructure; D051379:Mice; D008810:Mice, Inbred C57BL; D011897:Random Allocation; D013047:Specific Pathogen-Free Organisms; D013154:Spleen / Q000166:cytology / Q000528:radiation effects; D013950:Thymus Gland / Q000166:cytology / Q000528:radiation effects
D006657:Histones; C475961:gamma-H2AX protein, mouse