Crawford Bryan D, Henry Clarissa A, Clason Todd A, Becker Amanda L, Hille Merrill B
Department of Biology, University of Washington, Seattle, Washington 98195-1800, USA.
Mol Biol Cell. 2003 Aug;14(8):3065-81. doi: 10.1091/mbc.e02-08-0537. Epub 2003 Jun 13.
We investigated the focal adhesion proteins paxillin and Fak, and the cell-cell adhesion protein cadherin in developing zebrafish (Danio rerio) embryos. Cadherins are expressed in presomitic mesoderm where they delineate cells. The initiation of somite formation coincides with an increase in the phosphorylation of Fak, and the accumulation of Fak, phosphorylated Fak, paxillin, and fibronectin at nascent somite boundaries. In the notochord, cadherins are expressed on cells during intercalation, and phosphorylated Fak accumulates in circumferential rings where the notochord cells contact laminin in the perichordal sheath. Subsequently, changes in the orientations of collagen fibers in the sheath suggest that Fak-mediated adhesion allows longitudinal expansion of the notochord, but not lateral expansion, resulting in notochord elongation. Novel observations showed that focal adhesion kinase and paxillin concentrate at sites of cell-cell adhesion in the epithelial enveloping layer and may associate with actin cytoskeleton at epithelial junctions containing cadherins. Fak is phosphorylated at these epithelial junctions but is not phosphorylated on Tyr397, implicating a noncanonical mechanism of regulation. These data suggest that Fak and paxillin may function in the integration of cadherin-based and integrin-based cell adhesion during the morphogenesis of the early zebrafish embryo.
我们研究了发育中的斑马鱼(Danio rerio)胚胎中的粘着斑蛋白桩蛋白和粘着斑激酶,以及细胞间粘附蛋白钙粘蛋白。钙粘蛋白在前体节中胚层表达,在那里它们界定细胞。体节形成的起始与粘着斑激酶磷酸化的增加,以及新生体节边界处粘着斑激酶、磷酸化粘着斑激酶、桩蛋白和纤连蛋白的积累同时发生。在脊索中,钙粘蛋白在细胞插入过程中表达,磷酸化粘着斑激酶在脊索细胞与脊索周鞘中的层粘连蛋白接触的圆周环中积累。随后,鞘中胶原纤维方向的变化表明,粘着斑激酶介导的粘附允许脊索纵向扩展,但不允许横向扩展,从而导致脊索伸长。新的观察结果表明,粘着斑激酶和桩蛋白集中在上皮包被层的细胞间粘附部位,并可能与含有钙粘蛋白的上皮连接处的肌动蛋白细胞骨架相关联。粘着斑激酶在这些上皮连接处被磷酸化,但在Tyr397处未被磷酸化,这暗示了一种非经典的调节机制。这些数据表明,粘着斑激酶和桩蛋白可能在斑马鱼早期胚胎形态发生过程中基于钙粘蛋白和基于整合素的细胞粘附整合中发挥作用。