Fatma Abdelhedi1,2,3, Céline Chalas4, Jean-Maurice Petit5, Nouha Abid6, Elyes Mokadem7, Syrine Hizem8, Hassen Kamoun9, Leila Keskes9, Jean-Michel Dupont10,8. 1. Génomique, Epigénétique et Physiopathologie de la Reproduction, U1016 INSERM-UMR 8104 CNRS, Institut Cochin, Université Paris Descartes, Paris, France. abdelhedi_f@yahoo.fr. 2. Laboratoire de Cytogénétique, Hôpital Cochin, APHP, 75014, Paris, France. abdelhedi_f@yahoo.fr. 3. Laboratoire de Génétique Moléculaire Humaine, Faculté de Médecine de Sfax, Sfax, Tunisie. abdelhedi_f@yahoo.fr. 4. Laboratoire d'Histologie Embryologie - Biologie de la Reproduction, Hôpital Cochin, APHP, 75014, Paris, France. 5. Sorbonne Paris Cité, Université Paris Descartes, Paris, France. 6. Centre d'Aide Médicale à la Procréation, Polyclinique Ibn Nafiss, Sfax, Tunisie. 7. Centre d'Aide Médicale à la Procréation, Polyclinique El Alia, Sfax, Tunisie. 8. Laboratoire de Cytogénétique, Hôpital Cochin, APHP, 75014, Paris, France. 9. Laboratoire de Génétique Moléculaire Humaine, Faculté de Médecine de Sfax, Sfax, Tunisie. 10. Génomique, Epigénétique et Physiopathologie de la Reproduction, U1016 INSERM-UMR 8104 CNRS, Institut Cochin, Université Paris Descartes, Paris, France.
Abstract
PURPOSE: To explore the three-dimensional (3D) organization of sperm genome in DPY19L2-deficient globozoospermic patients speculating a link between DPY19L2 and genome organization of sperm nucleus. METHODS: This is a study of chromatin organization in DPY19L2-deficient globozoospermic patients and healthy donors using three-dimensional fluorescence in situ hybridization (3D-FISH) combined with confocal laser scanning microscopy followed by 3D image analysis. The 3D structures of sperm nuclei, chromocenter, telomeric regions and chromosome territories (CTs), were reconstructed using IMARIS software, and the relative radial position for each individual signal was calculated. Statistical analysis used a non-parametric Mann-Whitney test was appropriate with significance at p < 0.05. RESULTS: DPY19L2-deficient globozoospermic patients display impaired sperm chromocenter organization resulting in an increased number of chromocenters (5.4 vs 3.5; p < 0.0001). Moreover, radial positions of telomeres are modified with a more central position in globozoospermic nuclei. 3D-FISH analysis of five chromosome territories (CTs) (X, Y, 7, 17, 18) showed that DPY19L2-deficient globozoospermic sperm nuclei display altered spatial organization of CT X, CT 7 and CT 18. CONCLUSIONS: Our findings strengthen the hypothesis that DPY19L2 might be considered as a LINC-like protein having a crucial role in the organization of nuclear chromatin in sperm nucleus through its interaction with nuclear lamina. Our results might also explain defective embryonic development after intracytoplasmic sperm injection (ICSI) performed with DPY19L2-deficient globozoospermic sperm.
PURPOSE: To explore the three-dimensional (3D) organization of sperm genome in DPY19L2-deficient globozoospermic patients speculating a link between DPY19L2 and genome organization of sperm nucleus. METHODS: This is a study of chromatin organization in DPY19L2-deficient globozoospermic patients and healthy donors using three-dimensional fluorescence in situ hybridization (3D-FISH) combined with confocal laser scanning microscopy followed by 3D image analysis. The 3D structures of sperm nuclei, chromocenter, telomeric regions and chromosome territories (CTs), were reconstructed using IMARIS software, and the relative radial position for each individual signal was calculated. Statistical analysis used a non-parametric Mann-Whitney test was appropriate with significance at p < 0.05. RESULTS:DPY19L2-deficient globozoospermic patients display impaired sperm chromocenter organization resulting in an increased number of chromocenters (5.4 vs 3.5; p < 0.0001). Moreover, radial positions of telomeres are modified with a more central position in globozoospermic nuclei. 3D-FISH analysis of five chromosome territories (CTs) (X, Y, 7, 17, 18) showed that DPY19L2-deficient globozoospermic sperm nuclei display altered spatial organization of CT X, CT 7 and CT 18. CONCLUSIONS: Our findings strengthen the hypothesis that DPY19L2 might be considered as a LINC-like protein having a crucial role in the organization of nuclear chromatin in sperm nucleus through its interaction with nuclear lamina. Our results might also explain defective embryonic development after intracytoplasmic sperm injection (ICSI) performed with DPY19L2-deficient globozoospermic sperm.
Authors: M Hazzouri; S Rousseaux; F Mongelard; Y Usson; R Pelletier; A K Faure; C Vourc'h; B Sèle Journal: Mol Reprod Dev Date: 2000-03 Impact factor: 2.609
Authors: J Auger; F Eustache; A G Andersen; D S Irvine; N Jørgensen; N E Skakkebaek; J Suominen; J Toppari; M Vierula; P Jouannet Journal: Hum Reprod Date: 2001-12 Impact factor: 6.918
Authors: Ian K Greaves; Willem Rens; Malcolm A Ferguson-Smith; Darren Griffin; Jennifer A Marshall Graves Journal: Chromosome Res Date: 2003 Impact factor: 4.620