Abstract
Programmed cell death is a crucial process in neural development that affects mature neurons and glial cells, as well as proliferating precursors and recently born neurons at earlier stages. However, the regulation of the early phase of neural cell death and its function remain relatively poorly understood. In mouse models defective in homologous recombination or nonhomologous end-joining (NHEJ), which are both DNA double-strand break (DSB) repair pathways, there is massive cell death during neural development, even leading to embryonic lethality. These observations suggest that natural DSBs occur frequently in the developing nervous system. In this study, we have found that several components of DSB repair pathways are activated in the developing mouse retina at stages that coincide with the onset of neurogenesis. In short-term organotypic retinal cultures, we confirmed that the repair pathways can be modulated pharmacologically. Indeed, inhibiting the DNA-dependent protein kinase (DNA-PK) catalytic subunit, which is involved in NHEJ, with NU7026 increased caspase-dependent cell death and selectively reduced the neuron population. This observation concurs with an increase in the number of apoptotic neurons found after NU7026 treatment, as also observed in the embryonic scid mouse retina, a mutant that lacks DNA-PK catalytic subunit activity. Therefore, our results implicate the generation of DSB and DNA-PK-mediated repair in neurogenesis in the developing retina.
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Abbreviations
- ATM:
-
ataxia telangiectasia mutated
- ATR:
-
ataxia telangiectasia and Rad-3 related
- DNA-PK:
-
DNA-dependent protein kinase
- DSB:
-
double-strand break
- E:
-
embryonic day
- γH2AX:
-
Ser139 phosphorylated histone H2AX
- HR:
-
homologous recombination
- NHEJ:
-
nonhomologous end-joining
- P:
-
postnatal day
- RGC:
-
retinal ganglion cell
- RT:
-
room temperature
- SCID:
-
severe combined immunodeficiency
- TUNEL:
-
TdT-mediated dUTP nick end-labeling
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Acknowledgements
This work was supported by the Ministerio de Ciencia e Innovación, Spain (Grant SAF2007–66175 to EJdlR). JB is an FPI Fellow (Ministerio de Ciencia e Innovación). We thank Dr. Violeta Gómez-Vicente for critical reading of the paper, and Professor Flora de Pablo for continuous encouragement and ideas. We thank Maite Seisdedos, Noemí Alvarez, Ana Robles and the staff of the CIB animal house for technical support.
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Baleriola, J., Suárez, T. & de la Rosa, E. DNA-PK promotes the survival of young neurons in the embryonic mouse retina. Cell Death Differ 17, 1697–1706 (2010). https://doi.org/10.1038/cdd.2010.46
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DOI: https://doi.org/10.1038/cdd.2010.46
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