Multicolor Fluorescent Analysis in Zebrafish

描述（由申请人提供）：斑马鱼胚胎和幼体的透明性使其非常适合发育和生理过程的体内分析。本研究旨在开发斑马鱼细胞的荧光标记工具。先前的研究已经证明，随机DNA重组可以导致不同细胞中不同组合的荧光蛋白的表达。荧光蛋白的组合表达赋予细胞许多颜色，从而提供了一种区分相邻细胞和可视化细胞相互作用的方法。我们将开发工具，使这项技术广泛和容易地适用于斑马鱼发育和解剖学的研究（目标1），并生成下一代试剂和协议，使这项技术的复杂应用到斑马鱼生物学的各个方面（目标2）。 斑马鱼是一种脊椎动物模式生物，与人类共享许多生物过程。对斑马鱼的研究为了解出生缺陷和疾病的原因提供了重要的见解。建议的工作是通过开发工具来标记多种不同颜色的细胞并跟踪它们的发展，以提高该模型系统的实用性。

DESCRIPTION (provided by applicant): The translucency of zebrafish embryos and larvae makes them ideally suited for the in vivo analysis of developmental and physiological processes. This study aims to develop tools for the multicolor fluorescent labeling of zebrafish cells. Previous studies have demonstrated that stochastic DNA recombination can lead to the expression of different combinations of fluorescent proteins in different cells. The combinatorial expression of fluorescent proteins endows cells with many colors, thereby providing a way to distinguish adjacent cells and visualize cellular interactions. We will develop tools that make this technology widely and easily applicable to the study of zebrafish development and anatomy (Aim 1) and generate next-generation reagents and protocols that allow the sophisticated application of this technology to various aspects of zebrafish biology (Aim 2). PUBLIC HEALTH RELEVANCE: Zebrafish is a vertebrate model organism that shares many biological processes with humans. Research in zebrafish has provided important insights into the causes of birth defects and disease. The proposed work is to enhance the utility of this model system by developing tools to label cells in multiple different colors and follow their development.