Day and night variations in the repair of ionizing-radiation-induced DNA damage in mouse splenocytes.
In mammals, biological rhythms synchronize physiological and behavioral processes to the 24-h light-dark (LD) cycle. At the molecular level, self-sustaining processes, such as oscillations of transcription-translation feedback loops, control the circadian clock, which in turn regulates a wide variety of cellular processes, including gene expression and cell cycle progression. Furthermore, previous studies reported circadian oscillations in the repair capacity of DNA lesions specifically repaired by nucleotide excision repair (NER). However, it is so far only poorly understood if DNA repair pathways other than NER are under circadian control, in particular base excision and DNA strand break repair. In the present study, we analyzed potential day and night variations in the repair of DNA lesions induced by ionizing radiation (i.e., mainly oxidative damage and DNA strand breaks) in living mouse splenocytes using a modified protocol of the automated FADU assay. Our results reveal that splenocytes isolated from mice during the light phase (ZT06) displayed higher DNA repair activity than those of the dark phase (ZT18). As analyzed by highly sensitive and accurate qPCR arrays, these alterations were accompanied by significant differences in expression profiles of genes involved in the circadian clock and DNA repair. Notably, the majority of the DNA repair genes were expressed at higher levels during the light phase (ZT06). This included genes of all major DNA repair pathways with the strongest differences observed for genes of base excision and DNA double strand break repair. In conclusion, here we provide novel evidence that mouse splenocytes exhibit significant differences in the repair of IR-induced DNA damage during the LD cycle, both on a functional and on a gene expression level. It will be interesting to test if these findings could be exploited for therapeutic purposes, e.g. time-of-the-day-specific application of DNA-damaging treatments used against blood malignancies.
Molecular Toxicology Group, Department of Biology, University of Konstanz, D-78457 Konstanz, Germany.
Philipp
P
Palombo
Molecular Toxicology Group, Department of Biology, University of Konstanz, D-78457 Konstanz, Germany.
Maria
M
Moreno-Villanueva
Molecular Toxicology Group, Department of Biology, University of Konstanz, D-78457 Konstanz, Germany. Electronic address: aswin.mangerich@uni-konstanz.de.
Aswin
A
Mangerich
D000818:Animals; D002940:Circadian Rhythm / Q000502:physiology*; D004249:DNA Damage; D004260:DNA Repair / Q000235:genetics / Q000502:physiology*; D005786:Gene Expression Regulation; D051379:Mice; D012333:RNA, Messenger / Q000378:metabolism; D013154:Spleen / Q000166:cytology / Q000502:physiology* / Q000528:radiation effects; D014965:X-Rays
D012333:RNA, Messenger
Circadian rhythm; DNA repair; FADU assay; Gene profiling; Ionizing radiation; Molecular clock; qPCR