title: Dpy19l2-deficient globozoospermic sperm display altered genome packaging and DNA damage that compromises the initiation of embryo development.
abstract: We recently identified the DPY19L2 gene as the main genetic cause of human globozoospermia. Non-genetically characterized cases of globozoospermia were associated with DNA alterations, suggesting that DPY19L2-dependent globozoospermia may be associated with poor DNA quality. However the origins of such defects have not yet been characterized and the consequences on the quality of embryos generated with globozoospermic sperm remain to be determined. Using the mouse model lacking Dpy19l2, we compared several key steps of nuclear compaction. We show that the kinetics of appearance and disappearance of the histone H4 acetylation waves and of transition proteins are defective. More importantly, the nuclear invasion by protamines does not occur. As a consequence, we showed that globozoospermic sperm presented with poor sperm chromatin compaction and sperm DNA integrity breakdown. We next assessed the developmental consequences of using such faulty sperm by performing ICSI. We showed in the companion article that oocyte activation (OA) with globozoospermic sperm is very poor and due to the absence of phospholipase Cζ; therefore artificial OA (AOA) was used to bypass defective OA. Herein, we evaluated the developmental potential of embryos generated by ICSI + AOA in mice. We demonstrate that although OA was fully rescued, preimplantation development was impaired when using globozoospermic sperm. In human, a small number of embryos could be generated with sperm from DPY19L2-deleted patients in the absence of AOA and these embryos also showed a poor developmental potential. In conclusion, we show that chromatin compaction during spermiogenesis in Dpy19l2 KO mouse is defective and leads to sperm DNA damage. Most of the DNA breaks were already present when the sperm reached the epididymis, indicating that they occurred inside the testis. This result thus suggests that testicular sperm extraction in Dpy19l2-dependent globozoospermia is not recommended. These defects may largely explain the poor embryonic development of most mouse and human embryos obtained with globozoospermic sperm.
authors.affiliation: Université Grenoble Alpes, Grenoble, F-38000, France Equipe 'Andrologie, Génétique et Cancer' Laboratoire AGIM, CNRS FRE3405, La Tronche, F-38700, France.
authors.forename: Sandra
authors.initials: S
authors.lastname: Yassine
authors.affiliation: Université Grenoble Alpes, Grenoble, F-38000, France Equipe 'Andrologie, Génétique et Cancer' Laboratoire AGIM, CNRS FRE3405, La Tronche, F-38700, France.
authors.forename: Jessica
authors.initials: J
authors.lastname: Escoffier
authors.affiliation: Université Grenoble Alpes, Grenoble, F-38000, France Equipe 'Andrologie, Génétique et Cancer' Laboratoire AGIM, CNRS FRE3405, La Tronche, F-38700, France.
authors.forename: Guillaume
authors.initials: G
authors.lastname: Martinez
authors.affiliation: Université Grenoble Alpes, Grenoble, F-38000, France Equipe 'Andrologie, Génétique et Cancer' Laboratoire AGIM, CNRS FRE3405, La Tronche, F-38700, France CHU de Grenoble, UF de Génétique Chromosomique, Grenoble, F-38000, France.
authors.forename: Charles
authors.initials: C
authors.lastname: Coutton
authors.affiliation: Université Grenoble Alpes, Grenoble, F-38000, France Equipe 'Andrologie, Génétique et Cancer' Laboratoire AGIM, CNRS FRE3405, La Tronche, F-38700, France.
authors.forename: Thomas
authors.initials: T
authors.lastname: Karaouzène
authors.affiliation: Clinique des Jasmins, 23, Av. Louis BRAILLE, 1002 Tunis, Tunisia.
authors.forename: Raoudha
authors.initials: R
authors.lastname: Zouari
authors.affiliation: Université Grenoble Alpes, Grenoble, F-38000, France Laboratoire Lésions des Acides Nucléiques, CEA, INAC-SCIB, F-38000 Grenoble, France.
authors.forename: Jean-Luc
authors.initials: JL
authors.lastname: Ravanat
authors.affiliation: Aix-Marseille Université-Inserm UMR 910, Génétique médicale et Génomique Fonctionnelle, 13385 Marseille Cedex 5, France APHM Hôpital La Conception, Gynépôle, Laboratoire de Biologie de la Reproduction - CECOS, 13385 Marseille Cedex 5, France.
authors.forename: Catherine
authors.initials: C
authors.lastname: Metzler-Guillemain
authors.affiliation: Department of Veterinary and Animal Sciences, University of Massachusetts, Amherst, 661 North Pleasant Street, Amherst, MA 01003, USA.
authors.forename: Hoi Chang
authors.initials: HC
authors.lastname: Lee
authors.affiliation: Department of Veterinary and Animal Sciences, University of Massachusetts, Amherst, 661 North Pleasant Street, Amherst, MA 01003, USA.
authors.forename: Rafael
authors.initials: R
authors.lastname: Fissore
authors.affiliation: Université Grenoble Alpes, Grenoble, F-38000, France CHU de Grenoble, Centre d'AMP-CECOS, BP217, Grenoble Cedex 9, F-38043, France.
authors.forename: Sylviane
authors.initials: S
authors.lastname: Hennebicq
authors.affiliation: Université Grenoble Alpes, Grenoble, F-38000, France Equipe 'Andrologie, Génétique et Cancer' Laboratoire AGIM, CNRS FRE3405, La Tronche, F-38700, France CHU de Grenoble, UF de Biochimie et Génétique Moléculaire, Grenoble, F-38000, France.
authors.forename: Pierre F
authors.initials: PF
authors.lastname: Ray
authors.affiliation: Université Grenoble Alpes, Grenoble, F-38000, France Equipe 'Andrologie, Génétique et Cancer' Laboratoire AGIM, CNRS FRE3405, La Tronche, F-38700, France christophe.arnoult@ujf-grenoble.fr.
authors.forename: Christophe
authors.initials: C
authors.lastname: Arnoult
mesh_terms: D000818:Animals; D004249:DNA Damage / Q000235:genetics / Q000502:physiology; D005260:Female; D005455:Fluorescent Antibody Technique; D006801:Humans; D007248:Infertility, Male / Q000235:genetics / Q000378:metabolism; D008297:Male; D008565:Membrane Proteins / Q000172:deficiency* / Q000235:genetics; D051379:Mice; D018345:Mice, Knockout; D009865:Oocytes / Q000378:metabolism; D011479:Protamines / Q000378:metabolism; D013087:Spermatids / Q000378:metabolism; D013091:Spermatogenesis / Q000235:genetics / Q000502:physiology; D013094:Spermatozoa / Q000378:metabolism* / Q000502:physiology
chemical_list: C511317:DPY19L2 protein, human; D008565:Membrane Proteins; D011479:Protamines
keywords: DNA compaction; Dpy19l2; globozoospermia; male infertility; protamine