Netrin Glycosylation Influences Chemotaxis and Haptotaxis

Netrin 糖基化影响趋化性和趋触性

• 批准号: 10665243
• 负责人: Stephanie Gupton
• 金额: $ 19.44万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-04-15 至 2025-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10665243
• 关键词: Address; Adhesives; Affect; Anatomy; Architecture; Axon; Behavior; Behavioral; Biochemical; Biochemistry; Biological Assay; Cell membrane; Chemotaxis; Cues; Data; Development; Embryo; Environment; Fluorescence Resonance Energy Transfer; Future; Genes; Glycoproteins; Goals; Human; Image; Impairment; In Vitro; Invertebrates; Investigation; Mediating; Microfluidics; Modeling; Modification; Molecular; Morphogenesis; Mus; Mutation; NTN1 gene; Neurons; Proteomics; Protocols documentation; Signal Transduction; Solubility; Techniques; Total Internal Reflection Fluorescent; Work; automated analysis; body system; cell motility; developmental neurobiology; extracellular; genetic manipulation; glycosylation; high reward; high risk; in vivo; insight; mechanotransduction; morphogens; mouse model; mutant; netrin receptor; neural; novel; receptor; response; sensor; spatiotemporal

PROJECT SUMMARY Fidelity in directional motility is critical for development. This proposal addresses an outstanding question that remains a major impasse in the field of developmental neurobiology: How does a single extracellular morphogen, the secreted glycoprotein netrin-1, function as both a both a long-range diffusive cue vs a local, adhesive cue, and both an attractive cue and a repulsive cue. Netrin-1 is critical in development of many organ systems and branching morphogenesis from invertebrates to humans, where it promotes both attractive and repulsive motility. However, whether netrin-1 functions as a soluble, chemotactic cue and/or as an adhesive, haptotactic cue remains under debate. This is significant, as chemotactic cues can guide over long distances, whereas haptotactic cues guide locally, and thus they achieve appropriate development via fundamentally distinct means. We recently established protocols to separate and purify two distinctly glycosylated forms of netrin-1; we found that one of these forms functions as an attractive haptotactic cue; the other acts as a chemotactic guidance cue that elicits concentration-dependent attractive or repulsive responses. Here we will molecularly characterize these different forms of netrin-1 with glycoanalytics. We will examine how differentially glycosylated forms of netrin-1 evoke diverse responses during haptotaxis and chemotaxis, with the ultimate goal of developing novel mouse models to manipulation netrin glycosylation and thus solubility in vivo. We use biochemical assays and develop a novel tension sensor in the netrin receptor DCC to investigate how different presentations of netrin-1 alter signal transduction and mechanotransduction, respectively.

项目概要 定向运动的保真度对于发育至关重要。该提案解决了一个悬而未决的问题： 仍然是发育神经生物学领域的一个主要僵局：单个细胞外形态发生素如何， 分泌的糖蛋白 netrin-1，既充当远程扩散线索，又充当局部粘附线索， 既是有吸引力的暗示，又是令人厌恶的暗示。 Netrin-1 对于许多器官系统的发育至关重要 从无脊椎动物到人类的形态发生分支，它促进吸引和排斥运动。 然而，netrin-1 是否充当可溶性趋化信号和/或粘附性趋触信号 仍在争论中。这很重要，因为趋化信号可以长距离引导，而 触觉线索在局部进行指导，因此它们通过根本不同的方式实现适当的发展。 我们最近建立了分离和纯化两种明显糖基化形式的 netrin-1 的方案；我们发现 其中一种形式可作为有吸引力的触觉提示；另一个充当趋化引导信号 引起浓度依赖性的吸引或排斥反应。在这里我们将从分子角度表征 这些不同形式的 netrin-1 与糖分析。我们将研究糖基化形式的差异如何 netrin-1 在趋触性和趋化性过程中引起不同的反应，最终目标是开发新的 小鼠模型来操纵 netrin 糖基化，从而操纵体内溶解度。我们使用生化分析和 在 netrin 受体 DCC 中开发一种新型张力传感器，以研究 netrin-1 的不同表现形式如何 分别改变信号转导和机械转导。