Saka Yasushi, Smith J C
Wellcome Trust/Cancer Research UK Gurdon Institute, Department of Zoology, University of Cambridge, Cambridge CB2 1QR, UK.
Dev Biol. 2004 Sep 15;273(2):210-25. doi: 10.1016/j.ydbio.2004.05.032.
The product of the Drosophila gene tribbles inhibits cell division in the ventral furrow of the embryo and thereby allows the normal prosecution of gastrulation. Cell division is also absent in involuting dorsal mesoderm during gastrulation in Xenopus, and to ask whether the two species employ similar mechanisms to coordinate morphogenesis and the cell cycle, we isolated a putative Xenopus homologue of tribbles which we call Xtrb2. Extensive cDNA cloning identified long and short forms of Xtrb2, termed Xtrb2-L and Xtrb2-S, respectively. Xtrb2 is expressed maternally and in mesoderm and ectoderm at blastula and gastrula stages. Later, it is expressed in dorsal neural tube, eyes, and cephalic neural crest. Time-lapse imaging of GFP-tagged Xtrb2-L suggests that during cell division, it is associated with mitotic spindles. Knockdown of Xtrb2 by antisense morpholino oligonucleotides (MOs) disrupted synchronous cell divisions during blastula stages, apparently as a result of delayed progression through mitosis and cytokinesis. At later stages, tissues expressing the highest levels of Xtrb2 were most markedly affected by morpholino knockdown, with perturbation of neural crest and eye development.
果蝇基因tribbles的产物可抑制胚胎腹沟处的细胞分裂,从而使原肠胚形成过程正常进行。非洲爪蟾原肠胚形成期间内卷的背侧中胚层也不存在细胞分裂,为探究这两个物种是否采用相似机制来协调形态发生和细胞周期,我们分离出了一个假定的非洲爪蟾tribbles同源物,将其命名为Xtrb2。通过广泛的cDNA克隆鉴定出了Xtrb2的长、短两种形式,分别称为Xtrb2-L和Xtrb2-S。Xtrb2在母源期以及囊胚期和原肠胚期的中胚层和外胚层中表达。之后,它在背侧神经管、眼睛和头部神经嵴中表达。对绿色荧光蛋白标记的Xtrb2-L进行延时成像显示,在细胞分裂期间,它与有丝分裂纺锤体相关。用反义吗啉代寡核苷酸(MO)敲低Xtrb2会破坏囊胚期的同步细胞分裂,这显然是由于有丝分裂和胞质分裂进程延迟所致。在后期阶段,表达Xtrb2水平最高的组织受吗啉代敲低的影响最为明显,神经嵴和眼睛发育受到干扰。