title: The ability to organize sperm DNA into functional chromatin is acquired during meiotic maturation in murine oocytes.
abstract: Following fertilization of meiotically mature eggs, the chromatin of the sperm becomes biochemically and structurally remodeled within the egg cytoplasm. Despite the essential role of the paternal genome during embryogenesis, little is known of when the activities that regulate this chromatin remodeling appear during oogenesis. To determine whether these activities were acquired during meiotic maturation, we inseminated maturing oocytes of mice shortly after germinal vesicle breakdown. As previously shown, insemination at this stage did not activate the maturing oocytes, which became arrested at metaphase II. Immunofluorescent analysis revealed that at 1 hr postinsemination the sperm chromatin was dispersed and contained protamines but was devoid of core histones H2B and H3. At 4 hr postinsemination, both protamine and core histones were detectable on the sperm chromatin. By 8 hr postinsemination protamines were absent, and histones stained maximally. The appearance of immunoreactive histones was correlated with a morphological transition of the sperm chromatin from the dispersed to a condensed state, which suggests that the assembly of the histones reflected modification of the chromatin to a somatic-like state in which it was competent to respond to the metaphase-promoting factor activity of the oocyte. Both the assembly of histones and chromatin condensation were reversibly blocked when protein synthesis was inhibited, indicating that the remodeling process required proteins synthesized during maturation. Injection of core histones into protein synthesis-inhibited oocytes failed to induce condensation of the sperm chromatin, which implies that correct remodeling requires synthesis during maturation of nonhistone proteins. To test the functional capacity of remodeled sperm chromatin, maturing oocytes were inseminated, allowed to continue maturation for 17 hr and then parthenogenetically activated. Following activation, the sperm-derived chromatin as well as that of the oocyte became decondensed within pronuclei and underwent DNA replication, indicating that sperm chromatin remodeled in maturing oocyte cytoplasm was functionally normal. When the postinsemination incubation time was reduced to 11 hr; however, neither the female nor the male pronuclei underwent DNA replication, implying that factors synthesized late during maturation are required for DNA replication after activation. Taken together, these results indicate that the ability to organize sperm DNA into functional somatic-like chromatin develops in oocytes during meiotic maturation, requires proteins synthesized during maturation, and can be expressed independently of activation.
authors.affiliation: Department of Biology, McGill University, Montreal, Canada.
authors.forename: D W
authors.initials: DW
authors.lastname: McLay
authors.forename: H J
authors.initials: HJ
authors.lastname: Clarke
mesh_terms: D000818:Animals; D002467:Cell Nucleus / Q000378:metabolism; D002843:Chromatin / Q000737:chemistry / Q000502:physiology*; D004247:DNA / Q000096:biosynthesis / Q000502:physiology*; D004261:DNA Replication / Q000502:physiology; D005260:Female; D006657:Histones / Q000032:analysis / Q000378:metabolism; D008297:Male; D008540:Meiosis / Q000502:physiology*; D051379:Mice; D009865:Oocytes / Q000166:cytology / Q000502:physiology*; D014176:Protein Biosynthesis; D013084:Sperm-Ovum Interactions / Q000502:physiology*; D013094:Spermatozoa / Q000737:chemistry / Q000166:cytology / Q000502:physiology*
chemical_list: D002843:Chromatin; D006657:Histones; D004247:DNA