Xu Yong-Nan, Cui Xiang-Shun, Sun Shao-Chen, Jin Yong-Xun, Kim Nam-Hyung
Department of Animal Sciences, Chungbuk National University, Cheongju, South Korea.
J Exp Zool A Ecol Genet Physiol. 2011 Jul 1;315(6):349-57. doi: 10.1002/jez.682. Epub 2011 Mar 25.
The use of intracytoplasmic sperm injection (ICSI) in model animals is a powerful approach for the study of species-specific fertilization processes and multiploidy embryogenesis. In this study, we examined the fertilization process in mouse oocytes following injection of a single mouse or cat sperm, two mouse spermatozoa or mouse and cat spermatozoa. These treatments did not affect histone H3K9 acetylation or methylation, although the pattern of DNA methylation differed following the injection of cat sperm. Immunocytochemical staining revealed that sperm chromatin was normally incorporated with female mouse chromatin following any of the four injection scenarios. Furthermore, metaphase was successfully entered to reach a normal two-cell stage, and cell division could even persist to produce blastocyst stage embryos. In addition, both mouse and cat Pou5l and Nanog mRNA were expressed in the hybrid embryos. These results suggest that, although epigenetic modification of DNA is affected by the sperm injection treatment, fertilization and cleavage occur in a non-species-specific manner. In addition, despite abnormal division of the chromosomes, intra- and inter-species ICSI produced embryos that could develop into blastocysts.
在模式动物中使用胞浆内单精子注射（ICSI）是研究物种特异性受精过程和多倍体胚胎发生的有力方法。在本研究中，我们检测了向小鼠卵母细胞注射单个小鼠或猫精子、两个小鼠精子或小鼠与猫精子后的受精过程。这些处理并未影响组蛋白H3K9的乙酰化或甲基化，不过在注射猫精子后DNA甲基化模式有所不同。免疫细胞化学染色显示，在上述四种注射情况中的任何一种之后，精子染色质都能正常地与雌性小鼠染色质结合。此外，成功进入中期并发育至正常的二细胞阶段，细胞分裂甚至可以持续进行以产生囊胚期胚胎。另外，小鼠和猫的Pou5l和Nanog mRNA在杂交胚胎中均有表达。这些结果表明，尽管DNA的表观遗传修饰受精子注射处理的影响，但受精和卵裂以非物种特异性方式发生。此外，尽管染色体存在异常分裂，但种内和种间ICSI产生的胚胎能够发育成囊胚。
