Mechanisms of Adhesive Interactions

粘合剂相互作用的机制

• 批准号: 7648141
• 负责人: Steven Bernard Oppenheimer
• 金额: $ 10.81万
• 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/7648141
• 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.

由于其透明度，可及性，简单性和可用性，海胆的胚胎被广泛广泛 用于研究人类发育和疾病中也发生的细胞相互作用。这就是为什么大海 Hilchin Embryo NA被指定为NIH模型系统，这是本研究中使用的基本原理。我们的 长期目标是了解一个fuhctipaliy重要的细胞互动的分子基础，即 发育中的肠道（Archenteron）附着在Blastoeel的屋顶上。我们的工作假设， 基于我们发表的初步结果，是在arehenteron尖端处的次级Mesehchyme细胞 通过含有甘露糖/葡萄糖部分相关的配体与凝集素样的配体结合到Blastoeel屋顶的细胞 这些配体的受体。这些具体目的将检验以下假设：我们将确定：（1）如果 这些细胞相互作用的微分解成分通过甘露糖/gluoose部分结合在一起 受体； （2）如果我们从活着的胚胎中分离出的分子，则阻止活细胞相互作用 胚胎，可以纯化和表征； （3）如果这些分子的活性成分是特定的 糖和受体； （4）如果纯化的分子阻止了活胚中的细胞相互作用； （5）如果是 分子位于正确的发育时间中干燥的正确细胞类型上。 （6）如果掩盖了 活胚中的分子阻止细胞相互作用。 （7）如果发现在两种海洋中相似 顽童。这项研究具有创新性和新颖性，因为该系统完全可以访问生活中的探测 胚胎，并允许微分辨率隔离并访问细胞相互作用的成分 直接实验。在此修订中，重要性是我们通过阴离子交换HPLC隔离了分子 符合介导细胞相互作用的标准。两个主动的HPLC分数包含1（35 kD）和2（35 kD） 页面上的150 kd）乐队。毫克量的pufifi.ed分子很容易获得，将是 与已知的蛋白质；弗罗里各种基因组数据库进行了比较。 与公共卫生有关。这项研究将确定控制特定细胞的分子机制 使用NIH指定模型的精美可访问的海胆Erhbryo的相互作用。这些机制 在控制人类发展和 癌症扩散，在实验不太易于访问的人类系统中。