摘要
Electrofusion between blastula cells and unfertilized eggs in loach were investigated usingdielectrophoretic field where, under alternating sinusoidal electric field, blastula cells formed beads-like chain in close contact with the unfertilized egg and cell fusion occurred between eggs and thecells in tight contact with them. The nuclei ofblastula cells were brought into the cytoplasm of therecipient eggs, where they promoted the development of the fused eggs just like the zygote nuclei.But the development of the fused eggs was different from that of zygotes. Several nuclei might enterone and the same egg simultaneously and all of them could undergo division, resulting in severalblastomere after the first cleavage of the recipient egg. Before blastula stage, the embryo developingfrom the fused egg showcd irregular shape, but it was soon regulated and developed to a normalblastula which often continued its development into a normal individual. Cell/egg electrofusion cameto its highest fosion rate (80%) 8nd hatching rate (20%), with cell density at 1×10~3 cells/ml, Ca^(++)concentration at 10 mM, mannitol at 0.2 M and when the blastula cells were digested with 100μg/ml pronase E for 6-10 min at 20℃. The mechanism underlying development of electrofused eggsis discussed. As the result indicates, electrofusion might prove to be a promising biotechnology justas nuclear transplantation.
Electrofusion between blastula cells and unfertilized eggs in loach were investigated using dielectrophoretic field where, under alternating sinusoidal electric field, blastula cells formed beads- like chain in close contact with the unfertilized egg and cell fusion occurred between eggs and the cells in tight contact with them. The nuclei ofblastula cells were brought into the cytoplasm of the recipient eggs, where they promoted the development of the fused eggs just like the zygote nuclei. But the development of the fused eggs was different from that of zygotes. Several nuclei might enter one and the same egg simultaneously and all of them could undergo division, resulting in several blastomere after the first cleavage of the recipient egg. Before blastula stage, the embryo developing from the fused egg showcd irregular shape, but it was soon regulated and developed to a normal blastula which often continued its development into a normal individual. Cell/egg electrofusion came to its highest fosion rate (80%) 8nd hatching rate (20%), with cell density at 1×10~3 cells/ml, Ca^(++) concentration at 10 mM, mannitol at 0.2 M and when the blastula cells were digested with 100 μg/ml pronase E for 6-10 min at 20℃. The mechanism underlying development of electrofused eggs is discussed. As the result indicates, electrofusion might prove to be a promising biotechnology just as nuclear transplantation.
基金
This work was supported by grants from Chinese National High-Tech.Project.
关键词
囊胚细胞
未受精卵
泥鳅
电融合
Blastula Cell
Unfertilized Egg
Electrofusion
Loach
