title: Subcellular localization of phospholipase Cζ in human sperm and its absence in DPY19L2-deficient sperm are consistent with its role in oocyte activation.
abstract: We recently identified the DPY19L2 gene as the main genetic cause of human globozoospermia (70%) and described that Dpy19l2 knockout (KO) mice faithfully reproduce the human phenotype of globozoospermia making it an excellent model to characterize the molecular physiopathology of globozoospermia. Recent case studies on non-genetically characterized men with globozoospermia showed that phospholipase C, zeta (PLCζ), the sperm factor thought to induce the Ca(2+) oscillations at fertilization, was absent from their sperm, explaining the poor fertilization potential of these spermatozoa. Since 30% of globozoospermic men remain genetically uncharacterized, the absence of PLCζ in DPY19L2 globozoospermic men remains to be formally established. Moreover, the precise localization of PLCζ and the reasons underlying its loss during spermatogenesis in globozoospermic patients are still not understood. Herein, we show that PLCζ is absent, or its presence highly reduced, in human and mouse sperm with DPY19L2-associated globozoospermia. As a consequence, fertilization with sperm from Dpy19l2 KO mice failed to initiate Ca(2+) oscillations and injected oocytes remained arrested at the metaphase II stage, although a few human oocytes injected with DPY19L2-defective sperm showed formation of 2-pronuclei embryos. We report for the first time the subcellular localization of PLCζ in control human sperm, which is along the inner acrosomal membrane and in the perinuclear theca, in the area corresponding to the equatorial region. Because these cellular components are absent in globozoospermic sperm, the loss of PLCζ in globozoospermic sperm is thus consistent and reinforces the role of PLCζ as an oocyte activation factor necessary for oocyte activation. In our companion article, we showed that chromatin compaction during spermiogenesis in Dpy19l2 KO mouse is defective and leads to sperm DNA damage. Together, these defects explain the poor fertilization potential of DPY19L2-globozoospermic sperm and the compromised developmental potential of embryos obtained using sperm from patients with a deletion of the DPY19L2 gene.
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: Sandra
authors.initials: S
authors.lastname: Yassine
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: 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 Grenoble Institut des Neurosciences, INSERM U.836, F-38000 Grenoble, France.
authors.forename: Julie
authors.initials: J
authors.lastname: Delaroche
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: 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: Université Grenoble Alpes, Grenoble F-38000, France Grenoble Institut des Neurosciences, INSERM U.836, F-38000 Grenoble, France.
authors.forename: Karin
authors.initials: K
authors.lastname: Pernet-Gallay
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: 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 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: D000177:Acrosome / Q000378:metabolism; D000818:Animals; D005260:Female; D006801:Humans; D007248:Infertility, Male / Q000201:enzymology / Q000235:genetics; D008297:Male; D008565:Membrane Proteins / Q000172:deficiency* / Q000235:genetics; D051379:Mice; D018345:Mice, Knockout; D009865:Oocytes / Q000378:metabolism*; D013094:Spermatozoa / Q000201:enzymology* / Q000502:physiology*; D010738:Type C Phospholipases / Q000378:metabolism*
chemical_list: C511317:DPY19L2 protein, human; D008565:Membrane Proteins; D010738:Type C Phospholipases
keywords: DPY19L2; ICSI; acrosome; globozoospermia; male infertility; phospholipase C zeta