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. 2020 Mar 3;48(7):3678–3691. doi: 10.1093/nar/gkaa140

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© The Author(s) 2020. Published by Oxford University Press on behalf of Nucleic Acids Research.

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com

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Figure 3. — Atm inactivation in the Xrcc1^Nes-Cre brain results in abnormal cerebellum. (A) Gross view of the brains of mice from different genetic backgrounds (control, Atm^−/−, Xrcc1^Nes-Cre, Xrcc1^Nes-CreAtm^−/−) at P15. The red lines demarcate the cerebellum. The size of the cerebellum was reduced in the Xrcc1^Nes-CreAtm⁻^/- mice. OB, Olfactory bulb; CTX; Cerebral cortex; Ce, Cerebellum; Sp, Spinal cord. (B) Histopathological analysis of the cerebellum at P15. GAD (glutamate decarboxylase) immunoreactivity is for the GABAergic neuron network, and NF200 (neurofilament 200) immunoreactivity is for neurons. The Xrcc1^Nes-CreAtm^−/− cerebella showed signs of interneuron loss (yellow arrows, parvalbumin positive interneurons; white arrows, GABAergic neurons) in the molecular layer, similarly to the Xrcc1^Nes-Cre cerebella, suggesting that interneuron loss in Xrcc1 deficiency is ATM independent and p53 dependent (7). Gr, Granule cell layer; Pur, Purkinje cell layer; MO, Molecular layer. (C) Histopathological analysis of the cerebella of Atm^−/−, Xrcc1^Nes-Cre, and Xrcc1^Nes-CreAtm^−/− animals at P7 (sagittal plane), when the Purkinje cells are in a single-cell layer and foliation of all cerebellar lobules are noticeable. Nissl staining shows the overall structure of the cerebellar vermis. Calbindin immunoreactivity for the Purkinje cell layer, parvalbumin immunoreactivity for interneurons in the molecular layer and Purkinje cells, GABARa6 immunoreactivity for the granule cell layer, GFAP immunoreactivity for the Bergmann glia network, and PAX2 immunoreactivity for interneurons in the cerebellum are shown. The Atm null cerebellum was normal for the formation of Purkinje and granule cell layers and for the interneuron population. The Xrcc1^Nes-Cre cerebellum also showed normal development except for fewer interneurons (PAX2-positive cells) as described before (7). However, the Purkinje cell layer was not properly organized in the Xrcc1^Nes-CreAtm^−/− cerebellum, which was smaller and less foliated. Also, PAX2-positive interneurons were not rescued in the Xrcc1^Nes-CreAtm^−/− cerebellum. Gr, Granule cell layer; EGL, External germinal layer; Pur, Purkinje cell layer.