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Nhej1 Deficiency Causes Abnormal Development of the Cerebral Cortex

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Abstract

DNA double-strand breaks (DSBs) frequently occur in rapidly dividing cells such as proliferating progenitors during central nervous system development. If they cannot be repaired, these lesions will cause cell death. The non-homologous end joining (NHEJ) DNA repair pathway is the only pathway available to repair DSBs in post-mitotic neurons. The non-homologous end joining factor 1 (Nhej1) protein is a key component of the NHEJ pathway. Nhej1 interacts with Xrcc4 and Lig4 to repair DSBs. Loss of function of Xrcc4 or Lig4 is embryonic lethal in the mouse while the loss of Nhej1 is not. Surprisingly, the brains of Nhej1-deficient mice appear to be normal although NHEJ1 deficiency in humans causes severe neurological dysfunction and microcephaly. Here, we studied the consequences of Nhej1 dysfunction for the development of the cerebral cortex using in utero electroporation of inactivating small hairpin RNAs (shRNAs) in the developing rat brain. We found that decreasing Nhej1 expression during neuronal migration phases causes severe neuronal migration defects visualized at embryonic stages by an accumulation of heterotopic neurons in the intermediate zone. Knocked-down cells die by 7 days after birth and the brain regions where RNA interference was achieved are structurally abnormal, suffering from a reduction of the width of the external cortical layers. These results indicate that the Nhej1 protein is necessary for proper rat cortical development. Neurons unable to properly repair DNA DSBs are unable to reach their final destination during the development and undergo apoptosis, leading to an abnormal cortical development.

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Abbreviations

CP:

Cortical plate

DSBs:

Double-strand breaks

HR:

Homologous recombination

IZ:

Intermediate zone

LIG4:

Ligase IV

LV:

Lateral ventricle

NeuN:

Neuronal nuclear antigen

NHEJ:

Non-homologous end joining

NHEJ1:

Non-homologous end joining factor 1

TUNEL:

Terminal deoxynucleotidyltransferase-mediated dUTP end labeling

VZ:

Ventricular zone

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Acknowledgments

This work was supported by the Institut National de la Santé et de la Recherche Médicale (INSERM), Programme Hospitalier de Recherche Clinique (PHRC) from the French Ministry of Health and GIS Institut des Maladies Rares.

Conflict of Interest

The authors declare that they have no conflict of interest.

Author information

Authors and Affiliations

  1. Inserm UMR_S 910, 13385, Marseille, France

    Bilal El Waly, Marie-Reine Haddad & Laurent Villard

  2. Aix Marseille Université, GMGF, 13385, Marseille, France

    Bilal El Waly, Marie-Reine Haddad & Laurent Villard

  3. Inserm, U901, Marseille, France

    Emmanuelle Buhler

  4. INMED, Parc Scientifique de Luminy, Marseille, France

    Emmanuelle Buhler

  5. Plateforme postgénomique INMED-INSERM, Parc Scientifique de Luminy, Marseille, France

    Emmanuelle Buhler

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  1. Bilal El Waly
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  4. Laurent Villard
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Correspondence to Laurent Villard.

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El Waly, B., Buhler, E., Haddad, MR. et al. Nhej1 Deficiency Causes Abnormal Development of the Cerebral Cortex. Mol Neurobiol 52, 771–782 (2015). https://doi.org/10.1007/s12035-014-8919-y

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