Mechanisms of Adhesive Interactions

粘合剂相互作用的机制

• 批准号: 7455723
• 负责人: Steven Bernard Oppenheimer
• 金额: $ 10.9万
• 依托单位: CALIFORNIA STATE UNIVERSITY NORTHRIDGE
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/7455723
• 关键词: Adhesiveness; Adhesives; Animals; Anions; Binding; Biological Models; Cell Communication; Cells; DNA Sequence Rearrangement; Development; Disease; Embryo; Embryonic Development; Funding; Gastrocoele; Genome; Glucose; Goals; Grant; High Pressure Liquid Chromatography; Human; Human Development; Human Genome; Lead; Lectin; Letters; Life; Ligands; Localized; Malignant Neoplasms; Mannose; Masks; Mediating; Mesenchyme; Microdissection; Modeling; Molecular; Molecular Probes; Morphogenesis; National Center for Research Resources; Nose; Organism; Paper; Proteins; Public Health; Publishing; Research Institute; Right-On; Score; Sea Urchins; System; Testing; Time; Tissues; United States National Institutes of Health; Work; base; cell motility; cell type; genome database; innovation; man; milligram; novel; novel strategies; receptor; research study; sugar

Because of its transparency, accessibility, simplicity and availability, the sea urchin embryo has been widely used to study cellular interactions that also occur in human development and disease. That is why the sea urchin embryo nas been designated a NiH model system and isthe rationale for its use in this study. Our long-term goal is to understand the molecular basis of one fuhctipnaliy important cellularInteraction, namely the attachment of the developing gut (archenteron) to the roof of the blastoeoel. Our working hypothesis, based on our published preliminary results, is that secondary mesehchyme cells at the tip of the arehenteron bind to cells of the blastoeoel roof via ligands containing mannose/glucose moieties associatirig with lectin-like receptors for these ligands. These specific aims will test this hypothesis: We will determine: (1) if microdissected components of thes cellular interaction bind together via mannose/gluoose moieties and their receptors; (2) if molecules that we isolated from living embryos, that block the cellular interaction in live embryos, can be purified and characterized; (3) if the active components of these molecules are specific sugars and receptors; (4) if the purified molecules block the cellular interaction in living embryos; (5) if the molecules are localized on the right cell types arid at the right developmental time; (6) if masking the molecules in live embryos blocks the cellular interaction; (7) if the findings are similar in two species of sea urchins. This study is innovative and novel because the system is totally accessible to probes in living embryos and allows microdissection to isolate and access the components of the cellular interaction for direct experimentation. Of importance in this revision is that we isolated molecules by anion-exchange HPLC that meet criteria for mediating the cell interaction. Two active HPLC fractions contain 1 (35 kD) and 2 (35 kD and 150 kD) bands on PAGE. Milligram quantities of the pufifi.ed molecules are easily Obtainable and will be sequeheed and compared with known proteins;frorri various genome databases. Relevance to Public Health. This study will identify molecular mechanisms that control a specific cellular interaction using the exquisitely accessible sea urchin erhbryo, a NIH designated model. These mechanisms will be of major importance in the elucidation of similar mechanisms that control human development and cancer spread, in human systems that are not very accessible to experimentation.

由于海胆胚胎具有透明性、可获取性、简单性和可用性，已被广泛 用于研究人类发育和疾病中也发生的细胞相互作用。这就是为什么海 海胆胚胎被指定为 NiH 模型系统，这也是其在本研究中使用的基本原理。我们的 长期目标是了解一种非常重要的细胞相互作用的分子基础，即 发育中的肠道（archenteron）附着在囊胚顶部。我们的工作假设， 根据我们发表的初步结果，位于角锥体尖端的次级间充质细胞 通过含有甘露糖/葡萄糖部分的配体与凝集素样结合到囊胚顶细胞 这些配体的受体。这些具体目标将检验这个假设：我们将确定：（1）如果 这些细胞相互作用的显微解剖成分通过甘露糖/葡萄糖部分及其结合在一起 受体； (2) 如果我们从活体胚胎中分离出能够阻断活体胚胎中细胞相互作用的分子 胚胎，可以被纯化和表征； (3)这些分子的活性成分是否具有特异性 糖和受体； (4) 纯化的分子是否阻断活胚胎中的细胞相互作用； (5) 如果 分子定位在正确的细胞类型上并在正确的发育时间； (6) 如果屏蔽 活胚胎中的分子阻断细胞相互作用； (7) 如果两种海洋的结果相似 海胆。这项研究具有创新性和新颖性，因为该系统完全可以被生活中的探针所访问。 胚胎并允许显微解剖分离和获取细胞相互作用的成分 直接实验。本次修订中重要的是我们通过阴离子交换 HPLC 分离了分子 符合介导细胞相互作用的标准。两个活性 HPLC 级分包含 1 (35 kD) 和 2 (35 kD) 和 150 kD) PAGE 上的条带。毫克量的纯化分子很容易获得，并且可以 测序并与已知蛋白质进行比较；来自各种基因组数据库。 与公共卫生的相关性。这项研究将确定控制特定细胞的分子机制 使用 NIH 指定模型、易于操作的海胆 erhbryo 进行互动。这些机制 对于阐明控制人类发展和控制的类似机制将具有重要意义 癌症在不易进行实验的人体系统中扩散。