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X-Ray Induced DNA Damage and Repair in Germ Cells of PARP1−/− Male Mice
Unit of Radiation Biology and Human Health, Italian National Agency for New Technologies, Energy and Sustainable Economic Development (ENEA), CR Casaccia, Via Anguillarese 301, Roma 00123, Italy
Department of Ecology and Biology, University of Tuscia, Viterbo 01100, Italy
* Author to whom correspondence should be addressed.
Received: 24 June 2013; in revised form: 21 August 2013 / Accepted: 26 August 2013 / Published: 4 September 2013
Abstract: Poly(ADP-ribose)polymerase-1 (PARP1) is a nuclear protein implicated in DNA repair, recombination, replication, and chromatin remodeling. The aim of this study was to evaluate possible differences between PARP1−/− and wild-type mice regarding induction and repair of DNA lesions in irradiated male germ cells. Comet assay was applied to detect DNA damage in testicular cells immediately, and two hours after 4 Gy X-ray irradiation. A similar level of spontaneous and radiation-induced DNA damage was observed in PARP1−/− and wild-type mice. Conversely, two hours after irradiation, a significant level of residual damage was observed in PARP1−/− cells only. This finding was particularly evident in round spermatids. To evaluate if PARP1 had also a role in the dynamics of H2AX phosphorylation in round spermatids, in which γ-H2AX foci had been shown to persist after completion of DNA repair, we carried out a parallel analysis of γ-H2AX foci at 0.5, 2, and 48 h after irradiation in wild-type and PARP1−/− mice. No evidence was obtained of an effect of PARP1 depletion on H2AX phosphorylation induction and removal. Our results suggest that, in round spermatids, under the tested experimental conditions, PARP1 has a role in radiation-induced DNA damage repair rather than in long-term chromatin modifications signaled by phosphorylated H2AX.
Keywords: poly(ADP-ribose)polymerase-1; DNA repair; male mouse germ cells; comet assay; H2AX phosphorylation; ionizing radiation
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Villani, P.; Fresegna, A.M.; Ranaldi, R.; Eleuteri, P.; Paris, L.; Pacchierotti, F.; Cordelli, E. X-Ray Induced DNA Damage and Repair in Germ Cells of PARP1−/− Male Mice. Int. J. Mol. Sci. 2013, 14, 18078-18092.
Villani P, Fresegna AM, Ranaldi R, Eleuteri P, Paris L, Pacchierotti F, Cordelli E. X-Ray Induced DNA Damage and Repair in Germ Cells of PARP1−/− Male Mice. International Journal of Molecular Sciences. 2013; 14(9):18078-18092.
Villani, Paola; Fresegna, Anna M.; Ranaldi, Roberto; Eleuteri, Patrizia; Paris, Lorena; Pacchierotti, Francesca; Cordelli, Eugenia. 2013. "X-Ray Induced DNA Damage and Repair in Germ Cells of PARP1−/− Male Mice." Int. J. Mol. Sci. 14, no. 9: 18078-18092.