Wang Fay, Dumstrei Karin, Haag Thomas, Hartenstein Volker
Department of Molecular Cell and Developmental Biology, University of California Los Angeles, Los Angeles, CA 90095, USA.
Dev Biol. 2004 Jun 15;270(2):350-63. doi: 10.1016/j.ydbio.2004.03.002.
The Drosophila E-cadherin homolog, DE-cadherin, is expressed and required in all epithelial tissues throughout embryogenesis. Due to a strong maternal component of DE-cadherin, its early function during embryogenesis has remained elusive. The expression of a dominant negative DE-cadherin construct (UAS-DE-cad(ex)) using maternally active driver lines allowed us to analyze the requirements for DE-cadherin during this early phase of development. Maternally expressed DE-cad(ex) result in phenotype with variable expressivity. Most severely affected embryos have abnormalities in epithelialization of the blastoderm, resulting in loss of the blastodermal cells' apico-basal polarity and monolayered structure. Another phenotypic class forms a rather normal blastoderm, but shows abnormalities in proliferation and morphogenetic movements during gastrulation and neurulation. Mitosis of the mesoderm occurs prematurely before invagination, and proliferation in the ectoderm, normally a highly ordered process, occurs in a random pattern. Mitotic spindles of ectodermal cells, normally aligned horizontally, frequently occurred vertically or at an oblique angle. This finding further supports recent findings indicating that, in the wild-type ectoderm, the zonula adherens is required for the horizontal orientation of mitotic spindles. Proliferation defects in DE-cad(ex)-expressing embryos are accompanied by the loss of epithelial structure of ectoderm and neuroectoderm. These germ layers form irregular double or triple layers of rounded cells that lack zonula adherens. In the multilayered neuroectoderm, epidermal precursors, neuroblasts and ganglion mother cells occurred intermingled, attesting to the pivotal role of DE-cadherin in delamination and polarized division of neuroblasts. By contrast, the overall number and spacing of neuroblasts was grossly normal, indicating that DE-cadherin-mediated adhesion is less important for cell-cell interaction controlling the ratio of epidermal vs. neural progenitors.
果蝇E-钙黏蛋白同源物DE-钙黏蛋白在胚胎发育过程中所有上皮组织中表达且是必需的。由于DE-钙黏蛋白有强大的母体成分,其在胚胎发育早期的功能一直难以捉摸。利用母体活性驱动系表达显性负性DE-钙黏蛋白构建体(UAS-DE-cad(ex)),使我们能够分析发育早期阶段对DE-钙黏蛋白的需求。母体表达的DE-cad(ex)导致表型具有可变的表达性。受影响最严重的胚胎在胚盘上皮形成过程中出现异常,导致胚盘细胞顶-基极性和单层结构丧失。另一表型类别形成相当正常的胚盘,但在原肠胚形成和神经胚形成过程中显示出增殖和形态发生运动异常。中胚层的有丝分裂在内陷之前过早发生,而外胚层中的增殖(正常情况下是一个高度有序的过程)以随机模式发生。外胚层细胞的有丝分裂纺锤体通常水平排列,却频繁垂直或倾斜出现。这一发现进一步支持了最近的研究结果,即表明在野生型外胚层中,黏着小带对于有丝分裂纺锤体的水平定向是必需的。表达DE-cad(ex)的胚胎中的增殖缺陷伴随着外胚层和神经外胚层上皮结构的丧失。这些胚层形成不规则的双层或三层圆形细胞,缺乏黏着小带。在多层神经外胚层中,表皮前体细胞、神经母细胞和神经节母细胞相互混杂,证明了DE-钙黏蛋白在神经母细胞分层和极化分裂中的关键作用。相比之下,神经母细胞的总数和间距总体上是正常的,表明DE-钙黏蛋白介导的黏附对于控制表皮与神经祖细胞比例的细胞间相互作用不太重要。