Bottle Cells Are Required for the Initiation of Primary Invagination in the Sea Urchin Embryo

Abstract

Invagination of epithelial tissue occurs during gastrulation, neurulation, and organogenesis in many organisms. However, the underlying morphogenetic mechanisms of invagination are not understood. To elucidate these mechanisms, we have analyzed the initial invagination of the vegetal plate in the sea urchin embryo, a process termed primary invagination. At the onset of invagination, a ring of cells with highly constricted apices (bottle cells) encircles a group of two to eight round, central cells. To investigate the morphogenetic role of the bottle cells in the process of primary invagination, we have undertaken a series of laser ablation studies in which different proportions of various cell types were ablated and the effects were recorded using 4-D microscopy. Elimination of a 90°–180° arc of bottle cells markedly retards invagination, but only within the ablated region. Ablation of other cell types does not result in a statistically significant effect on primary invagination. These studies indicate that the number and arrangement of the bottle cells are critical factors for proper initiation of invagination. In addition, we have used the perturbing anti-hyalin antibody mAb183 to show that cell attachment to the hyaline layer is necessary for bottle cell formation and the initiation of primary invagination.