目的：为研究和了解细胞核与细胞质的相互作用。方法：采用3%蔗糖预处理显示及去核法进行昆明白小鼠与C57BL/6小鼠(黑色)之间卵母细胞MII期核(纺锤体)的移植、电融合、体外受精-胚胎移植(IVF-ET)。结果：经3%蔗糖预处理卵母细胞的去核率达100%。完成MII期核移植的重构卵母细胞约17.6%完成电融合。IVF的受精率为82%。受精卵2-细胞胚胎的发育率为80%～90%，8个1～4细胞胚胎移入两只自然怀孕1 d的寄母小鼠(Km×Km)输卵管，其中1只孕鼠生下3只小鼠，其中2只黑色小鼠为MII期核(纺锤体)移植生产的小鼠。移植产仔率为25%。结论：实验结果证明了3%蔗糖预处理显示MII期核(纺锤体)及去核效果可达100%；电融合的重构卵母细胞可完成正常的IVF。为进行不同年龄和卵龄间的MII期核移植提供了一个模型。%Objective: To study the interaction between nucleus and cytoplasm.rn Methods: Operation medium containing 3% sucrose was used to pretreatment murine oocytes and meiotic apparatus transfer(MAT) was carried out from C57BL/6 mouse (black) to enucleated Kunming murine oocytes and followed by electrofusion, in vitro fertilization (IVF) and embryo transfer.rn Results: The results showed that the enucleation rate by treating oocytes with 3% sucrose was 100%, 17.6% of reconstructed karyoplasm-recipient cytoplasm pairs fused sucessfully. When the fused oocytes were exposed to spermatozoa from C57BL/6 mice, 9 out of 11 (82%) were fertilized. Among them, eight embryos at one-to four-cell stages were transferred into the oviducts of two synchronous pregnant Kunming strain fosters and one delivered two normal C57BL/6 offsprings.rn Conclusion: This study indicated that MII meiotic apparatus or spindle sustains normal structure and function after micromanipulation and electrofusion. MAT provides a model for further research on the application of this technique to assistant human reproduction.
展开▼
