Dependence of DNA synthesis and in vitro development of bovine nuclear transfer embryos on the stage of the cell cycle of donor cells and recipient cytoplasts.
The effect of the stage of the cell cycle of donor cells and recipient cytoplasts on the timing of DNA replication and the developmental ability in vitro of bovine nuclear transfer embryos was examined. Embryos were reconstructed by fusing somatic cells with unactivated recipient cytoplasts or with recipient cytoplasts that were activated 2 h before fusion. Regardless of whether recipient cytoplasts were unactivated or activated, the embryos that were reconstructed from donor cells at the G0 phase initiated DNA synthesis at 6-9 h postfusion (hpf). The timing of DNA synthesis was similar to that of parthenogenetic embryos, and was earlier than that of the G0 cells in cell culture condition. Most embryos that were reconstructed from donor cells at the G1/S phase initiated DNA synthesis within 6 hpf. The developmental rate of embryos reconstructed by a combination of G1/S cells and activated cytoplasts was higher than the rates of embryos in the other combination of donor cells and recipient cytoplasts. The results suggest that the initial DNA synthesis of nuclear transfer embryos is affected by the state of the recipient oocytes, and that the timing of initiation of the DNA synthesis depends on the donor cell cycle. Our results also suggest that the cell cycles of somatic cells synchronized in the G1/S phase and activated cytoplasts of recipient oocytes are well coordinated after nuclear transfer, resulting in high developmental rates of nuclear transfer embryos to the blastocyst stage in vitro.
Kurosaka
Satoshi
S
Laboratory of Reproductive Biology, Graduate School of Agriculture, Kyoto University, Kyoto 606-8502, Japan.
Nagao
Yasumitsu
Y
Minami
Naojiro
N
Yamada
Masayasu
M
Imai
Hiroshi
H
D000818:Animals; D016590:Aphidicolin / Q000494:pharmacology; D015153:Blotting, Western; D002417:Cattle; D002453:Cell Cycle / Q000502:physiology*; D002467:Cell Nucleus / Q000235:genetics*; D016895:Culture Media, Serum-Free; D019926:Cyclin B / Q000096:biosynthesis; D056744:Cyclin B1; D003593:Cytoplasm / Q000378:metabolism*; D004247:DNA / Q000096:biosynthesis*; D004624:Embryo Transfer; D004622:Embryo, Mammalian / Q000378:metabolism; D005314:Embryonic and Fetal Development / Q000502:physiology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005260:Female; D016200:Maturation-Promoting Factor / Q000494:pharmacology; D008845:Microinjections; D010053:Ovary / Q000166:cytology; D012867:Skin / Q000166:cytology
D016895:Culture Media, Serum-Free; D019926:Cyclin B; D056744:Cyclin B1; D004791:Enzyme Inhibitors; D016590:Aphidicolin; D004247:DNA; D016200:Maturation-Promoting Factor