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

项目摘要 运动细胞对免疫系统至关重要，但异常的细胞运动是免疫系统的标志。 肿瘤的因此，细胞如何调节运动是一个重要的问题。在这个应用程序中，我们探索 在斑马鱼中通过实验操作细胞运动的遗传学， 细胞骨架-肌动蛋白捆绑蛋白L-plastin。L-纤维蛋白原通常只在白色血液中表达 细胞，但在许多类型的癌细胞中上调，并经常被研究为不利的肿瘤细胞。 这种疾病的预后标志物。然而，L-plastin在这些运动细胞中的基本作用 人口仍不清楚。对于这种蛋白质的研究，斑马鱼提供了最佳的选择 基因和细胞特征的结合。斑马鱼只有一个L-plastin拷贝 基因，便于与人类和小鼠中相应的基因座进行比较。基因工程在 斑马鱼现在是高效的，允许快速产生有效的敲除，敲入和 记者专线最后，斑马鱼的透明组织允许正常和 改变了几乎所有器官的细胞运动在我们以前工作的基础上，该项目将使用 斑马鱼L-质体蛋白敲除系，以检查该蛋白对细胞分裂，细胞粘附， 和免疫细胞迁移。这些研究将有助于我们了解L-plastin在 胚胎发生、免疫反应和肿瘤扩散--所有这些情况下，适当的细胞迁移都是至关重要的。 除了评估肌动蛋白捆绑、细胞骨架和细胞迁移之间的联系外， 拟议的工作将在我们的机构为未来的科学事业培养多样化的本科生。

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.