Maruyama Kouichi, Yasumasu Shigeki, Iuchi Ichiro
Life Science Institute, Sophia University, Kioi-cho Chiyoda-ku, Tokyo 102-8554, Japan.
J Biochem. 2002 Oct;132(4):581-9. doi: 10.1093/oxfordjournals.jbchem.a003260.
Using the teleost Oryzias latipes (medaka), we isolated three embryonic globin cDNAs (em.alpha-0, em.alpha-1, and em.beta-1) from the embryos 5 days after fertilization (at 30 degrees C) and two adult globin cDNAs (ad.alpha-1 and ad.beta-1) from the kidney of the fully-grown adult fish, and predicted their amino acid sequences. Molecular phylogenetic analysis showed that the embryonic globins were highly homologous in amino acid sequence to the embryonic globins previously identified in rainbow trout and zebrafish, and that they formed a monophyletic group among the teleostean globin molecules. They were clearly discriminated from the adult globin of the medaka. RT-PCR analysis showed that the embryonic globin mRNAs were intensely expressed in stage 30 and 38 embryos and in young fish 30 days after hatching. The level of expression decreased drastically after the young fish stage, and was low in fully-grown adult fish. The adult alpha globin mRNA ad.alpha-1 was scarcely expressed in the embryos, and the level of expression gradually increased in young to fully-grown adult fish. Unexpectedly, the adult beta globin mRNA ad.beta-1 was expressed throughout life, from the early embryonic stage to the fully-grown adult stage. This expression profile was quite different from that of the rainbow trout previously investigated. Some globins of the medaka were expressed both in primitive hematopoiesis and in definitive hematopoiesis.
我们以硬骨鱼青鳉为研究对象,从受精后5天(30摄氏度)的胚胎中分离出3种胚胎球蛋白cDNA(em.α - 0、em.α - 1和em.β - 1),并从成年青鳉的肾脏中分离出2种成年球蛋白cDNA(ad.α - 1和ad.β - 1),同时预测了它们的氨基酸序列。分子系统发育分析表明,这些胚胎球蛋白在氨基酸序列上与先前在虹鳟和斑马鱼中鉴定出的胚胎球蛋白高度同源,并且在硬骨鱼球蛋白分子中形成了一个单系群。它们与青鳉的成年球蛋白有明显区别。逆转录聚合酶链反应(RT-PCR)分析显示,胚胎球蛋白mRNA在30期和38期胚胎以及孵化后30天的幼鱼中强烈表达。幼鱼期后表达水平急剧下降,在成年青鳉中表达较低。成年α球蛋白mRNA ad.α - 1在胚胎中几乎不表达,其表达水平在幼鱼到成年青鳉中逐渐增加。出乎意料的是,成年β球蛋白mRNA ad.β - 1从胚胎早期到成年期都有表达。这种表达模式与先前研究的虹鳟有很大不同。青鳉的一些球蛋白在原始造血和定型造血过程中均有表达。