Development of goat embryos reconstituted with somatic cells: the effect of cell-cycle coordination between transferred nucleus and recipient oocytes.
The developmental ability and the nucleus and microtubule dynamics of nuclear transplanted goat embryos derived from in vitro matured oocytes were studied while controlling cell-cycle coordination of donor embryonic nuclei and recipient cytoplasts. Three groups of transfers were studied: G0/G1 (after the fibroblast cells grew to 100% confluence) and G2/M (nocodazole treated) phase fibroblasts transferred to MII cytoplasts (G0/G1-->MII and G2/M-->MII group, respectively), and G0/G1 phase fibroblasts transferred to preactivated cytoplasts, mostly at S-phase, (G0/G1-->Pre group) by electrical fusion. The results showed that fusion and developmental ability did not differ between G0/G1-->MII and G0/G1-->Pre groups. However the developmental rate of embryos in the G0/G1-->MII group was significantly higher than that of the G2/M-->MII group. Most fibroblast nuclei (G0/G1 and G2/M) transferred into MII oocytes underwent premature chromosome condensation (PCC). Normal spindle were only detected in the G0/G1-->MII group. In contract, fibroblast nuclei in pre-activated oocytes rarely underwent PCC, but formed a swollen nuclear structure. The data suggest that in vitro matured goat oocytes can support the development of somatic fibroblasts after nuclear transfer, G0/G1 -->MII and G0/G1-->S nuclear transfer might be effective ways for improving the developmental competence of the reconstituted embryos, and that G2/M-->MII nuclear transfer by electrical fusion (even in Ca2+-free fusion medium) induces abnormal chromosome ploidy.
Zhang
Li-Sheng
LS
State Key Laboratory of Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing, China.
Jiang
Man-Xi
MX
Lei
Zi-Li
ZL
Li
Rui-Chang
RC
Sang
Dan
D
Sun
Qing-Yuan
QY
Chen
Da-Yuan
DY
D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods*; D002453:Cell Cycle; D002467:Cell Nucleus / Q000378:metabolism*; D046149:Embryo Culture Techniques; D004622:Embryo, Mammalian / Q000166:cytology*; D005260:Female; D005347:Fibroblasts / Q000378:metabolism; D006041:Goats; D007150:Immunohistochemistry; D018613:Microscopy, Confocal; D008870:Microtubules / Q000378:metabolism; D009865:Oocytes / Q000166:cytology*; D008941:Spindle Apparatus; D013997:Time Factors