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

项目摘要 运动性细胞对免疫系统至关重要，但异常的细胞运动性是 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，我们将探索 通过实验在斑马鱼中操纵细胞运动性的遗传学 细胞骨架-肌动蛋白捆绑蛋白L-纤溶酶原蛋白。L-纤溶酶通常只在白血中表达 细胞，但在许多类型的癌细胞中上调，经常被研究为一种不利的 这种疾病的预后标志物。然而，L-纤溶酶在这些运动细胞中的基础作用 人口仍不清楚。对于这种蛋白质的研究，斑马鱼提供了一个最佳的 遗传和细胞特征的结合。斑马鱼只有一个L的复制品--纤溶酶 基因，便于与人类和小鼠的相应基因进行比较。中国的基因工程 斑马鱼现在效率很高，可以快速生产有效的敲除、敲入和 记者专线。最后，斑马鱼的透明组织可以实时显示正常和 改变了几乎所有器官中的细胞运动。在我们以前工作的基础上，这个项目将使用 斑马鱼L-血浆蛋白基因敲除株系检测该蛋白对细胞分裂、细胞黏附、 和免疫细胞的迁移。这些研究将有助于我们理解L-纤溶酶在心脏疾病中的作用。 胚胎发生、免疫反应和肿瘤扩散--所有这些领域，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， 拟议的工作将为我们机构未来的科学职业培养不同的本科生。

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.