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Parthenogenetic and nuclear transfer rabbit embryo development and apoptosis after activation treatments
Shu-Zhen Liu1, Man-Xi Jiang, Li-Ying Yan
1State Key Laboratory of Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing, China.
Abstract:
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.
Insights
Oocyte activation treatments significantly impact embryo development and reduce apoptosis in rabbit embryos. A specific protocol involving electrical pulses, inositol trisphosphate (IP3), and 6-dimethylaminopurine (6-DMAP) improved blastocyst rates and cell numbers while lowering apoptosis.
Area of Science:
- Reproductive Biology
- Developmental Biology
- Cell Biology
Background:
- Previous research focused on culture conditions affecting preimplantation embryo apoptosis.
- Apoptosis inhibitors were used on donor cells for nuclear transfer (NT) embryos, but activation treatments' effects remained unclear.
Purpose of the Study:
- To investigate the impact of various parthenogenetic activation (PA) treatments on rabbit embryo development, cell number, and apoptosis.
- To determine the optimal activation protocol for rabbit NT embryos to improve development and reduce apoptosis.
Main Methods:
- Evaluated multiple PA treatments on oocytes, assessing development, blastocyst cell count, and apoptosis.
- Identified the most effective PA treatment: electrical pulses, followed by inositol trisphosphate (IP3) electroporation, and 6-dimethylaminopurine (6-DMAP) exposure.
- Applied this optimized protocol to activate rabbit NT embryos.
Main Results:
- The optimal PA treatment (electrical pulses, IP3, 6-DMAP) resulted in a 72.7% blastocyst rate, 175 +/- 14.1 total cells, and 4.3 +/- 0.5% apoptosis.
- This protocol significantly outperformed other tested activation methods.
- Applying the protocol to NT embryos decreased blastocyst apoptosis and increased blastocyst rate and cell number.
Conclusions:
- Oocyte activation protocols critically influence not only embryo development and quality but also the level of embryo apoptosis.
- The specific activation method involving 6-DMAP is effective in improving rabbit NT embryo development and reducing apoptosis.
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