Hasegawa K, Kinoshita T
Developmental Biology, Faculty of Science, Kwansei Gakuin University, Nishinomiya, Hyogo, Japan.
Dev Growth Differ. 2000 Aug;42(4):337-46. doi: 10.1046/j.1440-169x.2000.00516.x.
Gastrulation is the most dynamic cell movement and initiates the body plan in amphibian development. In contrast to numerous molecular studies on mesodermal induction, the driving force of gastrulation is as yet poorly understood. A novel transmembrane protein, Xoom, was previously reported, which is required for Xenopus gastrulation. In the present study, the role of Xoom during Xenopus gastrulation was further examined in detail. Overexpression and misexpression of Xoom induced overproduction of Xoom protein, but not a changed phenotype. However, Xoom antisense ribonucleic acid (RNA) injection reduced the Xoom protein and caused gastrulation defects without any influence on the involution and translation levels of mesodermal marker genes. Normal migrating activity of dorsal mesodermal cells was recognized in the antisense RNA-injected explant. Morphological examination using artificial exogastrulation showed that convergent extension of mesodermal cells occurred normally, but the ectodermal cell layer significantly shrank in the antisense RNA-injected embryo. Comparison of cell shape among various experimental conditions showed that inhibition of cell spreading occurs specifically in the outer ectodermal layer of the antisense RNA-injected embryo. Cytochemical examination indicated disorganization of F-actin in the ectodermal cells of the antisense RNA-injected embryo. These results suggest that Xoom plays an important role in the epibolic movement of ectodermal cells through some regulation of actin filament organization.
原肠胚形成是最具活力的细胞运动,在两栖动物发育过程中启动身体蓝图。与众多关于中胚层诱导的分子研究不同,原肠胚形成的驱动力至今仍知之甚少。先前报道了一种新型跨膜蛋白Xoom,它是非洲爪蟾原肠胚形成所必需的。在本研究中,进一步详细研究了Xoom在非洲爪蟾原肠胚形成过程中的作用。Xoom的过表达和错误表达导致Xoom蛋白过量产生,但并未改变表型。然而,注射Xoom反义核糖核酸(RNA)可降低Xoom蛋白水平,并导致原肠胚形成缺陷,而对中胚层标记基因的内卷和翻译水平没有任何影响。在注射反义RNA的外植体中可观察到背侧中胚层细胞正常的迁移活性。使用人工外原肠胚形成进行的形态学检查表明,中胚层细胞的汇聚延伸正常发生,但在注射反义RNA的胚胎中外胚层细胞层明显收缩。比较各种实验条件下的细胞形状表明,细胞铺展的抑制特异性发生在注射反义RNA的胚胎的外胚层外层。细胞化学检查表明,注射反义RNA的胚胎外胚层细胞中的F-肌动蛋白紊乱。这些结果表明,Xoom通过对肌动蛋白丝组织的某种调节,在外胚层细胞的外包运动中发挥重要作用。