Parthenogenetic and nuclear transfer rabbit embryo development and apoptosis after activation treatments.
Previous studies mainly evaluated the effect of culture conditions on preimplantation embryo apoptosis. In order to inhibit apoptosis of nuclear transfer (NT) embryos, putative apoptosis inhibitors were used to treat donor cells. However, little is known about the effect of activation treatments on embryo apoptosis. We firstly investigated the effect of various parthenogenetic activation (PA) treatments on embryo development, blastocyst cell number, and apoptosis, and then one of these activation treatments proved to be most efficient was selected for activation rabbit NT embryos. The activation by electrical pulses and 30 min later, electroporation with 25 muM D-myoinositol 1,4,5-trisphosphate (IP3) in Ca(2+)- and Mg(2+)-free PBS, then exposure to 2.0 mM 6-dimethylaminopurine (6-DMAP) for 3 hr effectively activated rabbit oocytes, and resulted in significantly a higher blastocyst development rate (72.7%) and total cell number (175 +/- 14.1), and markedly lower apoptosis level of blastocyst (4.3 +/- 0.5) than all the other groups. When the same activation protocol was applied in NT embryo activation, we found that exposure of the embryos to 6-DMAP for 3 hr could decrease the apoptosis level of blastocyst and increase blastocyst rate and cell number. The results demonstrate that oocyte activation affects not only embryo development and quality but also embryo apoptosis.
Liu
Shu-Zhen
SZ
State Key Laboratory of Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing, China.
Jiang
Man-Xi
MX
Yan
Li-Ying
LY
Jiang
Yan
Y
Ouyang
Ying-Chun
YC
Sun
Qing-Yuan
QY
Chen
Da-Yuan
DY
D000225:Adenine / Q000031:analogs & derivatives / Q000494:pharmacology; D000818:Animals; D017209:Apoptosis / Q000187:drug effects / Q000502:physiology*; D001755:Blastocyst / Q000166:cytology / Q000502:physiology*; D002467:Cell Nucleus / Q000502:physiology; D002478:Cells, Cultured; D004627:Embryonic Induction / Q000187:drug effects / Q000502:physiology*; D053652:Nuclear Transfer Techniques; D009865:Oocytes / Q000166:cytology / Q000502:physiology*; D010312:Parthenogenesis / Q000502:physiology; D047428:Protein Kinase Inhibitors / Q000494:pharmacology; D011817:Rabbits
D047428:Protein Kinase Inhibitors; C001020:N(6),N(6)-dimethyladenine; D000225:Adenine