ANALYSIS OF THE CELL SURFACE IN MAMMALIAN DEVELOPMENT

成功的植入是成功发育的关键 哺乳动物胚胎的。滋养层细胞黏附作用的变化 在这一过程中发挥了重要作用。这里提出的实验将 探讨细胞表面在细胞分化中的作用 小鼠外胎盘锥体滋养层细胞的研究 其与子宫相互作用的分子机制 蜕膜细胞。先前的研究表明，不成熟的 滋养层细胞的黏附部分是由相互作用介导的 表面半乳糖基转移酶(GalTase)与糖偶联物 底物。向非粘附性侵袭性巨细胞分化 导致GalTase黏附系统下调， 尽管巨细胞仍具有表面半乳糖苷酶活性。这个 这里提出的实验将继续检验 GalTase在植入中。需要检验的假设是 GalTase的底物可利用性介导滋养层细胞过渡 从粘连到迁徙行为通过与 细胞外基质提供一种迁移机制。这些 实验将使用现有的或新产生的抗体来研究 细胞表面底物的发育调节，决定 GalTase在定义的矩阵上迁移中的作用，并检查 酶与底物的相互作用提供了一种 控制滋养层细胞分化的信号。植入将会 使用新开发的体外模型系统进行进一步研究。 子宫蜕膜细胞将被用作 EPC滋养层细胞的附着和铺展。这些文化将 被电子显微镜分析与黏附系统之间的 滋养层和蜕膜的胰酶分析，微扰， 和生化技术。将会产生特定的抗血清 与滋养层细胞有关的分子：蜕膜 互动。这些研究将提供有关 滋养层细胞的细胞和分子机制 分化和哺乳动物植入。此信息可以 应用于治疗不孕不育的技术开发 夫妇(试管受精和胚胎移植)以及提供新的 关于子宫组织侵袭的控制信息。这 信息可以用于研究无节制的入侵 与恶性细胞类型有关。

Successful implantation is critical for the successful development of the mammalian embryo. Changes in trophoblast cell adhesion play a major role in this process. The experiments proposed here will investigate the role of the cell surface in the differentiation of trophoblast in mouse ectoplacental cones (EPCs) and study the molecular mechanism involved in its interaction with uterine decidual cells. Previous work has indicated that immature trophoblast cell adhesion is mediated in part by the interaction of surface galactosyltransferase (GalTase) with glycoconjugate substrates. Differentiation to non-adhesive, invasive giant cells results in a downregulation of the GalTase adhesion system, although giant cells still possess surface GalTase activity. The experiments proposed here will continue to examine the role of GalTase in implantation. The hypothesis to be tested is that substrate availability for GalTase mediates trophoblast transition from adhesive to migratory behavior by interacting with extracellular substrates to provide a migratory mechanism. These experiments will use existing or newly produced antibodies to study developmental regulation of the cell surface substrate, determine the role of GalTase in migration on defined matrices, and examine the possibility that the enzyme:substrate interaction provides a signal controlling trophoblast differentiation. Implantation will be further studied using a newly developed in vitro model system. Uterine decidual cells will be used as a substrate for the attachment and spreading of EPC trophoblast. These cultures will be analyzed by electron microscopy and the adhesion systems between trophoblast and decidua analyzed with trypsinization, perturbation, and biochemical techniques. Specific antisera will be produced which identifies the molecules involved in trophoblast:decidua interactions. These studies will provide new information on the cellular and molecular mechanism involved in trophoblast differentiation and mammalian implantation. This information can be applied to the development of technologies to assist infertile couples (IVF and embryo transfer) as well as provide new information on the uterine control of tissue invasion. This information could be applied to the study of uncontrolled invasion associated with malignant cell type.