title: Topoisomerase II-mediated breaks in spermatozoa cause the specific degradation of paternal DNA in fertilized oocytes.
abstract: We have demonstrated that mouse spermatozoa can cleave their DNA into 50-kb fragments when treated with Triton X-100, MnCl(2), and CaCl(2). This cleavage, which is termed sperm chromatin fragmentation (SCF), is mediated by topoisomerase IIB (TOP2B) following stimulation by a factor in the epididymal fluid, most likely a nuclease, and can be at least partially religated by EDTA. When the protamines are removed, this DNA breakage is followed by digestion of the DNA by a nuclease(s). We tested whether the oocyte could repair TOP2B-induced sperm DNA breaks and whether partial religation by EDTA would allow spermatozoa to fertilize the oocytes normally. Oocytes injected with untreated spermatozoa developed normally. However, oocytes injected with spermatozoa treated with MnCl(2) and CaCl(2) to induce SCF, with or without subsequent EDTA treatment, failed to develop. In both of these treatment groups, the maternal pronuclei developed normally and replicated their DNA. However the paternal pronuclei did not replicate their DNA and this DNA began to disappear 6 h postinjection, which corresponded approximately to the time at which maternal DNA replication was initiated. These data suggest that when TOP2B is induced to cleave sperm DNA before fertilization, the paternal DNA is subsequently degraded by a highly regulated mechanism that does not affect the maternal chromatin. Furthermore, partial religation by EDTA of TOP2B-induced breaks prevents neither the inhibition of DNA synthesis nor DNA degradation.
authors.affiliation: Institute for Biogenesis Research, John A. Burns School of Medicine, University of Hawaii, Honolulu, Hawaii 96822, USA.
authors.forename: Yasuhiro
authors.initials: Y
authors.lastname: Yamauchi
authors.forename: Jeffrey A
authors.initials: JA
authors.lastname: Shaman
authors.forename: W Steven
authors.initials: WS
authors.lastname: Ward
mesh_terms: D000818:Animals; D004247:DNA / Q000378:metabolism*; D053960:DNA Breaks; D053938:DNA Fragmentation / Q000187:drug effects; D004261:DNA Replication; D004250:DNA Topoisomerases, Type II / Q000378:metabolism*; D004622:Embryo, Mammalian; D047108:Embryonic Development / Q000187:drug effects; D005260:Female; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008811:Mice, Inbred DBA; D017830:Octoxynol / Q000494:pharmacology; D009865:Oocytes / Q000187:drug effects / Q000378:metabolism*; D020554:Sperm Injections, Intracytoplasmic; D013094:Spermatozoa / Q000187:drug effects / Q000201:enzymology* / Q000378:metabolism
chemical_list: D017830:Octoxynol; D004247:DNA; D004250:DNA Topoisomerases, Type II