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The DNA double-strand “breakome” of mouse spermatids

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Abstract

De novo germline mutations arise preferentially in male owing to fundamental differences between spermatogenesis and oogenesis. Post-meiotic chromatin remodeling in spermatids results in the elimination of most of the nucleosomal supercoiling and is characterized by transient DNA fragmentation. Using three alternative methods, DNA from sorted populations of mouse spermatids was used to confirm that double-strand breaks (DSB) are created in elongating spermatids and repaired at later steps. Specific capture of DSB was used for whole-genome mapping of DSB hotspots (breakome) for each population of differentiating spermatids. Hotspots are observed preferentially within introns and repeated sequences hence are more prevalent in the Y chromosome. When hotspots arise within genes, those involved in neurodevelopmental pathways become preferentially targeted reaching a high level of significance. Given the non-templated DNA repair in haploid spermatids, transient DSBs formation may, therefore, represent an important component of the male mutation bias and the etiology of neurological disorders, adding to the genetic variation provided by meiosis.

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Acknowledgements

We are grateful to Isabelle Marois for technical assistance with flow cytometry; Julien Massonneau and Olivier Simard for technical help with DBrIC and Sébastien Rodrigue and Dominick Matteau for the preparation of sequencing library. This work was supported by a grant from the Canadian Institutes of Health Research (#MOP-136925) to G.B. and a scholarship from Fonds de recherche du Québec—Santé and the Réseau Québécois en Reproduction to M.-C.G.

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  1. Department of Biochemistry, Faculty of Medicine and Health Sciences, Université de Sherbrooke, Sherbrooke, QC, Canada

    Marie-Chantal Grégoire, Frédéric Leduc, Tiphanie Cavé, Mélina Arguin & Guylain Boissonneault

  2. Department of Biology, Faculty of Sciences, Université de Sherbrooke, Sherbrooke, QC, Canada

    Martin H. Morin & Pierre-Étienne Jacques

  3. Department of Medicine, Faculty of Medicine and Health Sciences, Université de Sherbrooke, Sherbrooke, QC, Canada

    Martin Richter

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  1. Marie-Chantal Grégoire
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M-CG, FL and GB designed the experiments; M-CG, FL, TC and MA performed the experiments; M-CG, P-EJ and MM performed bioinformatic analyses; MR performed flow cytometry. M-CG and GB wrote the manuscript; all authors read and commented on the manuscript.

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Correspondence to Guylain Boissonneault.

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Grégoire, MC., Leduc, F., Morin, M.H. et al. The DNA double-strand “breakome” of mouse spermatids. Cell. Mol. Life Sci. 75, 2859–2872 (2018). https://doi.org/10.1007/s00018-018-2769-0

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