Hypomethylation of paternal DNA in the late mouse zygote is not essential for development.
Global demethylation of DNA which marks the onset of development occurs asynchronously in the mouse; paternal DNA is demethylated at the the zygote stage, whereas maternal DNA is demethylated later in development. The biological function of such asymmetry and its underlying mechanisms are currently unknown. To test the hypothesis that the early demethylation of male DNA may be associated with protamine-histone exchange, we ,used round spermatids, whose DNA is still associated with histones, for artificial fertilization (round spermatid injection or ROSI), and compared the level of methylation of metaphase chromosomes in the resulting zygotes with the level of methylation in zygotes obtained after fertilization using mature sperm heads (intracytoplasmic sperm injection or ICSI). In contrast to ICSI-derived zygotes, ROSI-derived zygotes possessed only slightly demethylated paternal DNA. Both types of zygotes developed to term with similar rates which shows that hypomethylation of paternal DNA at the zygotic metaphase is not essential for full development in mice. Incorporation of exogenously expressed histone H2BYFP into paternal pronuclei was significantly higher in ICSI-derived zygotes than in ROSI-derived zygotes. Surprisingly, in the latter the incorporation of histone H2BYFP into the paternal pronucleus was still significantly higher than into the maternal pronucleus, suggesting that some exchange of chromatin-associated proteins occurs not only after ICSI but also after ROSI. This may explain why after ROSI, some transient demethylation of paternal DNA occurs early after fertilization, thus providing support for the hypothesis regarding the link between paternal DNA demethylation and protamine/histone exchange.
Polanski
Zbigniew
Z
Department of Developmental Biology, Max Planck Institute of Immunobiology, Freiburg, Germany. pola@iz.uj.edu.pl
Motosugi
Nami
N
Tsurumi
Chizuko
C
Hiiragi
Takashi
T
Hoffmann
Steffen
S
D000818:Animals; D002467:Cell Nucleus; D004247:DNA / Q000235:genetics*; D019175:DNA Methylation; D005260:Female; D005306:Fertilization; D005307:Fertilization in Vitro; D006657:Histones / Q000378:metabolism*; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D011479:Protamines / Q000378:metabolism; D013077:Sperm Head / Q000502:physiology; D020554:Sperm Injections, Intracytoplasmic; D013094:Spermatozoa / Q000502:physiology*; D015053:Zygote / Q000502:physiology*
D006657:Histones; D011479:Protamines; D004247:DNA