卵裂球电脉冲融合制作昆明小鼠四倍体胚胎

Forming Tetraploid Embryos with Electric Fusing Blastomeres of Kunming Mouse Embryos

为制作四倍体小鼠胚胎 ,用 4种脉冲电压强度 (6 0 0、 80 0、 10 0 0和 12 0 0V/cm )和 3种脉冲宽度(10、 30和 5 0 μs)组合 ,对 2 -细胞昆明小鼠胚胎做了融合实验。 6 0 0V/cm× 5 0 μs和 12 0 0V/cm× 30 μs两种组合融合率最高 ,分别为 91 6 %和 93 0 % ;而 6 0 0V/cm× 10 μs和 12 0 0V/cm× 5 0 μs时均显著低于其他各种组合 ,融合率分别为 5 8 9%和 6 0 2 %。以上结果表明 ,一定的脉冲电压强度是胚胎融合的必要条件 ,而脉冲宽度是影响胚胎融合及其后期发育的关键因素。激光共聚焦显微镜对融合后两细胞核的观察暗示 ,细胞核的迁移。

To form tetraploid embryos,electric pulses were used to stimulate 2 cell blastomeres of Kunming mice.The pulses included combinations of four strengths (600,800,1?000 and 1?200?V/cm) and three durations (10,30 and 50?μs).Of the combinations,600?V/cm×50?μs and 1?200?V/cm×30?μs produced the fusion rates of 91 6% and 93 0% respectively,and both were higher than the others.On the other hand,the pulses of 600?V/cm×10?μs and 1?200?V/cm×50?μs did the rates of 58 9% and 60 2% respectively,being significantly lower than the others.The results showed that a certain pulse strength was necessary to achieve embryo fusion,and the duration was a key factor affecting the embryo fusion and its later development.What observed with a laser confocal microscope implied that the allocation,approach and fusion of nucleus was a spontaneous process in fused embryos.