Follow That Cell: Motility Analysis of L-plastin Mutant Zebrafish

追踪该细胞：L-plastin 突变斑马鱼的运动分析

• 批准号: 10046864
• 负责人: ELIZABETH E LECLAIR
• 金额: $ 15.49万
• 依托单位: DE PAUL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-09-01 至 2022-03-25
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10046864
• 关键词: Actins; Adhesions; Adult; Affect; Alleles; Animal Model; Animals; Antibodies; Award; B-Lymphocytes; Biochemical; Biological Assay; Biological Markers; Biology; Biomedical Research; Bundling; CRISPR/Cas technology; Caenorhabditis elegans; Cancer Biology; Candida albicans; Cell Adhesion; Cell Cycle; Cell division; Cell surface; Cells; Cellular Structures; Cellular biology; Chemicals; Collaborations; Complement; Cytoskeleton; Data; Defect; Development; Disease; Disease Marker; Doctor of Philosophy; Educational process of instructing; Embryo; Embryonic Development; Environment; F-Actin; Fibroblasts; Focal Adhesions; Funding; Future; Genes; Genetic; Genetic Engineering; Health; Hematopoietic; Human; Image; Immune; Immune response; Immune system; In Vitro; Institution; Integrins; Knock-in; Knock-out; L-Plastin; LCP1 gene; Label; Leukocytes; Link; Malignant Neoplasms; Mammals; Mentors; Movement; Mus; National Institute of General Medical Sciences; Nematoda; Normal Cell; Organ; Orthologous Gene; Performance; Pharmacology; Phenotype; Population; Production; Prognostic Marker; Progress Reports; Proteins; Publications; Publishing; Reagent; Regulation; Reporter; Research; Role; Signal Transduction; Stains; Structure; Students; Study models; T-Lymphocyte; Tail; Techniques; Testing; Tissues; Training; Transformed Cell Line; Transgenic Organisms; United States National Institutes of Health; Universities; Visualization; Work; Yeasts; Zebrafish; base; blastomere structure; cancer cell; cancer type; career; career networking; cell behavior; cell motility; cell type; cost; genetic manipulation; in vivo; insight; macrophage; migration; monocyte; mutant; neoplastic; neoplastic cell; neutrophil; novel; overexpression; pathogen; plastin; public health relevance; response; survivorship; tumor; undergraduate student; vertebrate embryos; wound

PROJECT ABSTRACT Motile cells are critical for the immune system, yet aberrant cell motility is the hallmark of tumors. How cells regulate movement is therefore an important issue. In this application, we explore the genetics of cell motility by experimentally manipulating in zebrafish a key component of the cytoskeleton– the actin-bundling protein L-plastin. L-plastin is normally expressed only in white blood cells, but is upregulated in many types of cancer cells and is frequently studied as an unfavorable prognostic marker for this disease. However the fundamental role of L-plastin in these motile cell populations remains unclear. For the study of this protein, the zebrafish provides an optimal combination of genetic and cellular features. Zebrafish have only one copy of the L-plastin gene, facilitating comparison with the corresponding locus in humans and mice. Genetic engineering in zebrafish is now highly efficient, allowing the rapid production of effective knock-out, knock-in and reporter lines. Finally, the transparent tissues of zebrafish allow live visualization of both normal and modified cell movements in almost any organ. Building on our previous work, this project will use zebrafish L-plastin knockout lines to examine the impact of this protein on cell division, cell adhesion, and immune cell migration. These studies will contribute to our understanding of L-plastin function in embryogenesis, immune response and tumor spread-- all contexts where proper cell migration is critical. In addition to evaluating the links between actin bundling, the cytoskeleton, and cell migration, the proposed work will train diverse undergraduates at our institution for future scientific careers.

项目摘要 运动细胞对于免疫系统至关重要，但是异常细胞运动是 肿瘤。因此，细胞如何调节运动是一个重要的问题。在此应用程序中，我们探索 通过在斑马鱼中操纵细胞运动的遗传学 细胞骨架 - 肌动蛋白捆绑蛋白L-倍蛋白。 l-普拉斯汀通常仅以白血表示 细胞，但在许多类型的癌细胞中进行了更新，并且经常研究为不利 这种疾病的预后标记。但是，l-铂在这些母细胞中的基本作用 人口仍然不清楚。为了研究该蛋白质，斑马鱼提供了最佳 遗传和细胞特征的组合。斑马鱼只有一份l-plastin的副本 基因，支持与人类和小鼠相应的基因座的比较。基因工程 斑马鱼现在高效，可以快速生产有效的敲除，敲入和 记者线。最后，斑马鱼的透明组织允许对正常和 几乎所有器官中的细胞运动修饰。在我们以前的工作的基础上，该项目将使用 斑马鱼L-普拉斯汀敲除线，以检查该蛋白质对细胞分裂的影响，细胞粘合剂， 和免疫池迁移。这些研究将有助于我们理解L-普拉斯汀功能 胚胎发生，免疫响应和肿瘤扩散 - 适当细胞迁移至关重要的所有情况。 除了评估肌动蛋白束，细胞骨架和细胞迁移之间的联系外， 拟议的工作将在我们机构的未来科学职业上培训潜水员的本科生。