Abstract
Genetic approaches have provided evidence that DNA end-joining problems serve an essential role in neuronal survival during development of mammalian embryos. In the present study, we tested whether the DNA repair enzyme, DNA dependent protein kinase, plays an important role in the survival of cerebral cortical neurons in mice. DNA-PK is comprised of a DNA-binding subunit called Ku and a catalytic subunit called DNA-PKcs. In mice with the scid mutation, DNA-PKcs is truncated near the kinase domain, which causes loss of kinase activity. We compared the spatial and temporal aspects of neuronal cell death in scid versus isogenic wild-type embryos and found a significant increase in dying cells in scid mice, as assessed by nuclear changes, DNA fragmentation and caspase-3 activity. Additional biochemical and immunocytochemical studies indicated that of several DNA repair enzymes investigated, only PARP was increased in scid mice, possibly in response to elevated DNA strand breaks. Cell Death and Differentiation (2001) 8, 245–255
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Abbreviations
- ATM:
-
ataxia telangiectasia mutated
- CP:
-
cortical plate
- caspase:
-
cysteine dependent aspartate-specific protease
- DNA-PK:
-
DNA dependent protein kinase, DNA-PKcs, DNA dependent protein kinase catalytic subunit
- dsDNA:
-
double stranded DNA
- DTT:
-
dithiothreitol
- HR:
-
homologous recombination
- IZ:
-
intermediate zone
- NHEJ:
-
non-homologous end joining
- PARP:
-
poly (ADP) ribose polymerase
- PCD:
-
programmed cell death
- PMSF:
-
phenylmethylsulfonyl fluoride
- pNA:
-
para-nitroaniline
- RT:
-
room temperature
- scid :
-
severe combined immunodeficiency
- VZ:
-
ventricular zone
- WT:
-
wild-type
- XRCC4:
-
X-ray repair cross complementation-4
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Acknowledgements
The authors thank members of the Naegele lab including: Magda Chechlacz, Karen Gilliams-Francis, Joshua Gooley, Andrew Scott, Matt Daugherty, and Jenn Weiss for scientific discussions. In addition, we would like to thank Aruna Vemuri for technical assistance, Lisa Gagnon for animal husbandry and Paul Lombroso for helpful comments on the manuscript. This work has been supported by NSF grant IBN-9600974.
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Vemuri, M., Schiller, E. & Naegele, J. Elevated DNA double strand breaks and apoptosis in the CNS of scid mutant mice. Cell Death Differ 8, 245–255 (2001). https://doi.org/10.1038/sj.cdd.4400806
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DOI: https://doi.org/10.1038/sj.cdd.4400806
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