Kaftanovskaya Elena, Motosugi Nami, Kinoshita Masato, Ozato Kenjiro, Wakamatsu Yuko
Laboratory of Freshwater Fish Stocks, Bioscience and Biotechnology Center, Nagoya University, Furo-cho, Chikusa-ku, Nagoya 464-8601, Japan.
Dev Growth Differ. 2007 Dec;49(9):691-8. doi: 10.1111/j.1440-169X.2007.00962.x. Epub 2007 Sep 16.
Chromosomal abnormalities such as ploidy mosaicism have constituted a major obstacle to the successful nuclear transfer of adult somatic cell nuclei in lower vertebrates to date. Euploid mosaicism has been reported previously in well-developed amphibian transplants. Here, we investigated ploidy mosaicisms in well-developed transplants of adult somatic cell nuclei in medaka fish (Oryzias latipes). Donor nuclei from primary cultured cells from the adult caudal fin of a transgenic strain carrying the green fluorescent protein gene (GFP) were transferred to recipient nonenucleated eggs of a wild-type strain to produce 662 transplants. While some of the transplants developed beyond the body formation stage and several hatched, all exhibited varying degrees of abnormal morphology, limited growth and subsequent death. Twenty-one transplants, 19 embryos and two larvae, were selected for chromosomal analysis; all were well-developed 6-day-old or later embryonic stages exhibiting slight morphological abnormalities and the same pattern of GFP expression as that of the donor strain. In addition, all exhibited various levels of euploid mosaicism with haploid-diploid, haploid-triploid or haploid-diploid-triploid chromosome sets. No visible chromosomal abnormalities were observed. Thus, euploid mosaicism similar to that observed in amphibians was confirmed in well-developed nuclear transplants of fish.
迄今为止,诸如倍性嵌合体之类的染色体异常一直是低等脊椎动物中成年体细胞细胞核成功进行核移植的主要障碍。先前在发育良好的两栖类移植中已报道过整倍体嵌合体。在此,我们研究了青鳉(Oryzias latipes)中成年体细胞细胞核发育良好的移植中的倍性嵌合体。将携带绿色荧光蛋白基因(GFP)的转基因品系成年尾鳍原代培养细胞的供体细胞核转移到野生型品系的受体无核卵中,以产生662个移植体。虽然一些移植体发育到了身体形成阶段之后,有几个孵化了,但所有移植体都表现出不同程度的异常形态、生长受限以及随后死亡。选择了21个移植体、19个胚胎和2个幼虫进行染色体分析;所有这些都是发育良好的6日龄或更晚的胚胎阶段,表现出轻微的形态异常以及与供体品系相同的绿色荧光蛋白表达模式。此外,所有样本都表现出不同水平的整倍体嵌合体,具有单倍体 - 二倍体、单倍体 - 三倍体或单倍体 - 二倍体 - 三倍体染色体组。未观察到明显的染色体异常。因此,在发育良好的鱼类核移植中证实了与两栖类中观察到的相似的整倍体嵌合体。