P1-SPATIAL REGULATION OF PROTEIN KINASE A SIGNALING DURING GROWTH CONE GUIDANCE

生长锥引导过程中蛋白激酶 A 信号传导的 P1-空间调节

• 批准号: 7725300
• 负责人: Alan K Howe
• 金额: $ 23.44万
• 依托单位: UNIVERSITY OF VERMONT & ST AGRIC COLLEGE
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-07-01 至 2009-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7725300
• 关键词: Actins; Axon; Behavior; Chemotaxis; Co-Immunoprecipitations; Complex; Computer Retrieval of Information on Scientific Projects Database; Coupling; Cues; Cyclic AMP-Dependent Protein Kinases; DCC gene; Data; Development; Disruption; Event; Family; Funding; Grant; Growth Cones; Institution; Ligands; Mediating; Molecular; Morphology; Nature; Neurons; Phosphorylation; Protein Kinase; Regulation; Research; Research Personnel; Resources; Signal Transduction; Source; Synapses; Testing; United States National Institutes of Health; axon guidance; cell motility; extracellular; neuronal growth; receptor; response

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. During development, extracellular guidance cues regulate actin cytoskeletal dynamics in neuronal growth cones to control axon outgrowth and trajectory towards proper synaptic targets. Among these cues are the netrins, a family of secreted molecules capable of chemoattraction, chemorepulsion, and regulation of branching behavior. The response to netrin signaling is mediated principally through the DCC (Deleted in Colorectal Cancers) receptor family. Delineation of the signals downstream of DCC and the mechanisms through which they couple to cytoskeletal dynamics is fundamentally important to our understanding of netrin-mediated developmental events. We have previously shown that the cAMP-dependent protein kinase (PKA) is spatially regulated during chemotactic cell movement and that subcellular anchoring of PKA is required for efficient chemotaxis. Our preliminary data show that expression of DCC promotes activation of PKA in a ligand-dependent fashion. This activity correlates with phosphorylation of the Mena/VASP family of cytoskeletal regulators, an effect abrogated by disruption of PKA anchoring, suggesting the involvement of a specific PKA-containing signaling complex. We have also observed that co-immunoprecipitation of PKA and DCC is disrupted by inhibition of PKA anchoring. These data support a hypothesis in which association with, and activation of, PKA by DCC is involved in coupling netrin/DCC signaling to cytoskeletal regulation during axon guidance. We will test this by (1) elucidating the nature of the interaction between DCC and PKA, (2) investigating the molecular consequences of DCC association with and signaling through PKA, and (3) investigating the functional consequences of DCC/PKA signaling on neuronal morphology, dynamics, and guidance.

该子项目是利用该技术的众多研究子项目之一 资源由 NIH/NCRR 资助的中心拨款提供。子项目及 研究者 (PI) 可能已从 NIH 的另一个来源获得主要资金， 因此可以在其他 CRISP 条目中表示。列出的机构是 中心，不一定是研究者的机构。 在发育过程中，细胞外引导信号调节神经元生长锥中的肌动蛋白细胞骨架动力学，以控制轴突的生长和朝向适当突触目标的轨迹。这些线索包括网络蛋白，这是一类能够进行化学吸引、化学排斥和分支行为调节的分泌分子。对 netrin 信号传导的反应主要通过 DCC（结直肠癌删除）受体家族介导。 DCC 下游信号的描述以及它们与细胞骨架动力学耦合的机制对于我们理解 netrin 介导的发育事件至关重要。我们之前已经证明，cAMP 依赖性蛋白激酶 (PKA) 在趋化细胞运动过程中受到空间调节，并且 PKA 的亚细胞锚定是有效趋化作用所必需的。我们的初步数据表明 DCC 的表达以配体依赖性方式促进 PKA 的激活。这种活性与细胞骨架调节因子 Mena/VASP 家族的磷酸化相关，破坏 PKA 锚定会消除这种作用，表明含有特定 PKA 的信号复合物参与其中。我们还观察到，PKA 锚定的抑制会破坏 PKA 和 DCC 的免疫共沉淀。这些数据支持这样一种假设，即 DCC 与 PKA 的关联和激活参与了轴突引导过程中 netrin/DCC 信号传导与细胞骨架调节的耦合。我们将通过（1）阐明 DCC 和 PKA 之间相互作用的性质，（2）研究 DCC 与 PKA 关联并通过 PKA 信号传导的分子后果，以及（3）研究 DCC/PKA 信号传导对神经元的功能后果来测试这一点。形态、动力学和引导。