title: Further evidence that sperm nuclear proteins are necessary for embryogenesis.
abstract: We have recently presented evidence that the structural integrity of the mouse sperm nuclear matrix may be necessary for the proper unpackaging of sperm DNA for participation in embryogenesis. It is likely that the sperm nuclear matrix contributes to the organisation of the sperm DNA and its disturbance can seriously damage the paternal genome or its expression. In this work, we confirm our previous data and further suggest that even very subtle changes in the sperm nuclear structure may have a significant impact on embryo development. As reported previously, dithiothreitol (DTT) in the presence of an ionic detergent, ATAB, destabilized the nuclear matrix as measured by the halo assay, and oocytes injected with these nuclei failed to develop. We also discovered that omitting the protease inhibitor PMSF from the buffers used to extract spermatozoa prevented sperm injected into oocytes from participating in development. The organization of DNA into loop domains by the nuclear matrix in these nuclei appeared normal, as measured by the halo assay. Oocytes injected with sperm nuclei that had been washed with ATAB in the presence of phenylmethylsulphonyl fluoride (PMSF) but in the absence of DTT resulted in live births. Neither DTT treatment nor the absence of PMSF would be expected to disrupt the integrity of the paternal DNA. The data therefore suggest that even very subtle alterations in the structural proteins of the nucleus are enough to deprive sperm DNA of the ability to contribute to embryonic development.
authors.affiliation: Division of Urology, Robert Wood Johnson Medical School, New Brunswick, New Jersey 08903, USA.
authors.forename: W S
authors.initials: WS
authors.lastname: Ward
authors.forename: H
authors.initials: H
authors.lastname: Kishikawa
authors.forename: H
authors.initials: H
authors.lastname: Akutsu
authors.forename: H
authors.initials: H
authors.lastname: Yanagimachi
authors.forename: R
authors.initials: R
authors.lastname: Yanagimachi
mesh_terms: D000818:Animals; D002467:Cell Nucleus / Q000502:physiology* / Q000648:ultrastructure; D004247:DNA / Q000737:chemistry; D003902:Detergents / Q000494:pharmacology; D004229:Dithiothreitol / Q000494:pharmacology; D004624:Embryo Transfer; D005314:Embryonic and Fetal Development / Q000502:physiology*; D005260:Female; D008297:Male; D051379:Mice; D008815:Mice, Inbred Strains; D008854:Microscopy, Electron; D009687:Nuclear Proteins / Q000502:physiology*; D010664:Phenylmethylsulfonyl Fluoride / Q000494:pharmacology; D000644:Quaternary Ammonium Compounds / Q000494:pharmacology; D020554:Sperm Injections, Intracytoplasmic; D013094:Spermatozoa / Q000166:cytology / Q000502:physiology* / Q000648:ultrastructure
chemical_list: D003902:Detergents; D009687:Nuclear Proteins; D000644:Quaternary Ammonium Compounds; D010664:Phenylmethylsulfonyl Fluoride; D004247:DNA; D004229:Dithiothreitol