Sakai C, Konno F, Nakano O, Iwai T, Yokota T, Lee J, Nishida-Umehara C, Kuroiwa A, Matsuda Y, Yamashita M
Laboratory of Molecular and Cellular Interactions, Faculty of Advanced Life Science, Hokkaido University, Sapporo, 060-0810, Japan.
Chromosome Res. 2007;15(6):697-709. doi: 10.1007/s10577-007-1155-9. Epub 2007 Jun 26.
An interspecific hybrid medaka (rice fish) between Oryzias latipes and O. hubbsi is embryonically lethal. To gain an insight into the cellular and molecular mechanisms that cause the abnormalities occurring in the hybrid medaka, we investigated the behavior of chromosomes and the expression patterns of proteins responsible for the chromosome behavior. The number of chromosomes in the hybrid embryos gradually decreased to nearly half, since abnormal cell division with lagging chromosomes at anaphase eliminated the chromosomes from the cells. The chromosome lagging occurred at the first cleavage and continued throughout embryogenesis even after the midblastula transition. Fluorescent in-situ hybridization analyses revealed that the chromosomes derived from O. hubbsi are preferentially eliminated in both O. latipes-hubbsi and O. hubbsi-latipes embryos. Whole-mount immunocytochemical analyses using antibodies against alpha-tubulin, gamma-tubulin, inner centromere protein, Cdc20, Mad2, phospho-histone H3 and cohesin subunits (SMC1alpha, SMC3 and Rad21) showed that the expression patterns of these proteins in the hybrid embryos are similar to those in the wild-type embryos, except for phospho-histone H3. Phospho-histone H3 present on chromosomes at metaphase was lost from normally separated chromosomes at anaphase, whereas it still existed on lagging chromosomes at anaphase, indicating that the lagging chromosomes remain in the metaphase state even when the cell has proceeded to the anaphase state. On the basis of these findings, we discuss the cellular and molecular mechanisms of chromosome elimination in the hybrid medaka.
青鳉属的日本青鳉(Oryzias latipes)和赫氏青鳉(O. hubbsi)之间的种间杂交青鳉胚胎致死。为深入了解导致杂交青鳉出现异常的细胞和分子机制,我们研究了染色体行为以及负责染色体行为的蛋白质的表达模式。杂交胚胎中的染色体数量逐渐减少至近一半,因为后期出现染色体滞后的异常细胞分裂会将染色体从细胞中消除。染色体滞后现象在第一次卵裂时就出现,并在整个胚胎发育过程中持续,即使在囊胚中期转换之后也是如此。荧光原位杂交分析表明,在日本青鳉 - 赫氏青鳉和赫氏青鳉 - 日本青鳉胚胎中,源自赫氏青鳉的染色体都优先被消除。使用针对α - 微管蛋白、γ - 微管蛋白、着丝粒内蛋白、Cdc20、Mad2、磷酸化组蛋白H3和黏连蛋白亚基(SMC1α、SMC3和Rad21)的抗体进行的整体免疫细胞化学分析表明,除了磷酸化组蛋白H3外,这些蛋白质在杂交胚胎中的表达模式与野生型胚胎中的相似。中期染色体上存在的磷酸化组蛋白H3在后期正常分离的染色体上消失了,而在后期滞后染色体上仍然存在,这表明即使细胞已经进入后期状态,滞后染色体仍处于中期状态。基于这些发现,我们讨论了杂交青鳉中染色体消除的细胞和分子机制。