DSU Sub-Project 2

DSU 子项目 2

• 批准号: 8731950
• 负责人: Cynthia M. van Golen
• 金额: $ 16.63万
• 依托单位: DELAWARE STATE UNIVERSITY
• 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/8731950
• 关键词: AMD3100; Actin-Binding Protein; Actins; Axon; Binding; CXCL12 gene; CXCR4 Receptors; CXCR4 gene; Cell Line; Cells; Cellular Morphology; Centers of Research Excellence; Chemicals; Chemotaxis; Confocal Microscopy; Cues; Data; Defect; Delaware; Development; Extracellular Matrix; Human; Immigration; Immunoprecipitation; In Vitro; Insulin-Like Growth Factor I; Insulin-Like-Growth Factor I Receptor; Laboratories; Length; Ligands; Mass Spectrum Analysis; Mediating; Mediator of activation protein; Morphology; Nervous system structure; Neural Crest; Neural Crest Cell; Neurites; Neuroblastoma; Neurons; Neurosciences Research; Play; Population; Post-Translational Protein Processing; Process; Proteins; Regulation; Role; Signal Pathway; Synapses; Testing; Work; axonal guidance; base; cell motility; chemokine receptor; immunocytochemistry; in vivo; inhibitor/antagonist; migration; neuroblastoma cell; neutralizing antibody; polymerization; receptor; receptor function; research study

The external cues that guide the migration of developing neurons and outgrowth of neurons upon differentiation have been intensely studied for decades. CXCR4, a chemokine receptor, has been implicated in the regulation of chemotaxis, neuronal migration, and axonal guidance. Neuroblastoma cells, which are of neural crest origin, are capable of differentiating into more mature sympathetic neurons in culture, and insulin-like growth factor I (IGF-1), has been shown to promote both migration and neurite outgrowth in these cells. In addition, neuroblastoma cells often express high levels of CXCR4, are responsive to CXCLI 2 and, depending upon the level of differentiation, are capable of producing fully extended axons. The mediator of the cytoskeletal changes seen in during process extension is actin polymerization. We have shown that in the neuroblastoma cell line, SHSY-5Y, both CXCR4 and IGF I receptors are involved in neuronal outgrowth, however, treatment with ligands for these receptors results in different cellular morphologies. CXCR4 stimulation stimulated the cells to take on a more differentiated neuronal form and directly involved actin, while IGF-IR stimulation resulted in a very immature neuronal morphology with shorter, broader processes. Based on these results, our overall hypothesis is that CXCR4 promotes neuronal migration and neurite extension through direct regulation of actin dynamics. Our preliminary data suggest that activation of CXCR4 by CXCL12 in cultured neuroblastoma cells promotes the elongation of neurites, and we have found CXCR4 along these projections. In this work we will be testing three specific hypotheses: 1) Cellular context, including the extracellular matrix present and the concentration of CXCL12 ligand to which the cells are exposed play a large role in determining the signaling pathways that are activated, and thereby the ability of the cells to migrate; 2) CXCR4 activation by CXCL12 promotes an increase in neurite length in neuroblastoma cells; 3) CXCR4 regulates elongation of neuronal processes through interaction with actin using the actinbinding protein Dbn; We will use immunocytochemistry/confocal microscopy, neurite analysis, and immunoprecipitation with cultured neuroblastoma and sympathetic neurons to test these hypotheses. Posttranslational modification of CXCR4 will be analyzed using Mass Spectroscopy.

引导发育中的神经元迁移和分化后神经元生长的外部线索已经被深入研究了几十年。CXCR4是一种趋化因子受体，参与趋化性、神经元迁移和轴突引导的调节。神经母细胞瘤细胞起源于神经嵴，能够在培养中分化为更成熟的交感神经元，而胰岛素样生长因子I （IGF-1）已被证明可以促进这些细胞的迁移和神经突的生长。此外，神经母细胞瘤细胞通常表达高水平的CXCR4，对cxcr2有反应，并且根据分化水平，能够产生完全延伸的轴突。在过程延伸过程中看到的细胞骨架变化的介质是肌动蛋白聚合。