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.

项目摘要 定向运动的保真度对于发展至关重要。该提案解决了一个杰出的问题 在发育神经生物学领域仍然是主要的僵局：单个细胞外形态学如何，如何 分泌的糖蛋白Netrin-1作为远程扩散提示既是局部的粘合剂提示 都是有吸引力的提示和排斥的提示。 Netrin-1对于许多器官系统的开发至关重要 将形态发生从无脊椎动物到人类，在那里它促进了吸引力和排斥运动。 但是，Netrin-1是否充当可溶性，趋化提示和/或作为粘合剂，触觉提示 仍在辩论中。这很重要，因为趋化提示可以在长距离上引导，而 Haptotactic提示在本地指导，因此它们通过根本不同的手段实现了适当的发展。 我们最近建立了分离和净化Netrin-1的两种明显糖基化形式的方案；我们发现 这些形式之一是一种有吸引力的触觉提示。其他充当趋化引导提示 这会引起浓度依赖性的有吸引力或排斥反应。在这里，我们将分子表征 这些不同形式的Netrin-1与甘氨酸分析。我们将研究如何差异化糖基化形式 Netrin-1在触觉和趋化性期间引起了多种反应，其最终目标是开发新颖的反应 小鼠模型操纵Netrin糖基化，从而在体内溶解度。我们使用生化测定法和 在Netrin受体DCC中开发一种新型的张力传感器，以研究Netrin-1的不同演示 分别更改信号转导和机械转导。