Patterning of the Primitive Heart Tube in Zebrafish

描述(由申请人提供)：胚胎心脏的形成和/或功能异常通常会导致胚胎死亡或在以后的生活中出现严重的健康问题。在细胞和分子水平上了解心脏早期发育的基本机制不仅具有科学挑战性，而且具有临床意义。我们选择斑马鱼作为我们研究早期心脏模式的模式生物，因为它服从于遗传、胚胎和分子操作。此外，所有脊椎动物的胚胎心脏都经历了相似的形态发生过程，并受到保守的遗传回路的调节。从一种模式生物获得的信息可能也适用于其他脊椎动物，包括人类。在这个项目中，我们采用三种独立的方法来研究原始心脏管形态发生的细胞和分子机制。我们最近对斑马鱼心脏和心智突变的研究发现，Na，K-ATPase a1B1在原始心管的延长中发挥了意想不到的作用。我们将探讨Na，K-ATPase A1、B1功能丧失对心肌细胞构筑的影响。我们还将在细胞和遗传水平上研究Na，K-ATPase A1，B1和其他对斑马鱼原始心管形态发生至关重要的基因之间的相互作用(目标1)。此外，我们将研究Na，K-ATPase A1 B1调节斑马鱼和培养细胞中原始心管延长的分子机制，作为了解参与原始心管形成的遗传网络的第一步(特定目标2)。最后，从正在进行的斑马鱼基因筛查中，我们发现了一个影响原始心管图案的新突变。我们将表征该突变的表型，并确定导致该突变表型的分子损伤(目标3)。本项目提出的细胞、分子和遗传学相结合的研究将为原始心管形态发生的机制提供新的和深入的见解。

DESCRIPTION (provided by applicant): Abnormalities in the formation and/or function of the embryonic heart often lead to embryonic lethality or severe health problems later in life. Understanding the fundamental mechanisms underlying early heart development at the cellular and molecular level is not only scientifically challenging but is also clinically relevant. We choose the zebrafish as our model organism to study early cardiac patterning because it is amenable to genetic, embryological and molecular manipulation. Furthermore, the embryonic hearts of all vertebrate species undergo similar morphogenic processes and are regulated by conserved genetic circuits. Information obtained from one model organism may apply to other vertebrates, including humans. In this project, we take three independent approaches to study the cellular and molecular mechanisms governing primitive heart tube morphogenesis. Our recent study on the zebrafish heart and mind mutation uncovered an unexpected role of Na,K-ATPase a1B1 in the elongation of the primitive heart tube. We will investigate the impact of loss of function of Na,K-ATPase a1 B1 on the cellular architecture of cardiomyocytes. We will also investigate the interaction of Na,K-ATPase a1 B1 other genes critical for the morphogenesis of the primitive heart tube in zebrafish at the cellular and genetic level (Aim 1). Furthermore we will investigate the molecular mechanism by which the Na,K-ATPase a1 B1 regulates the elongation of the primitive heart tube in the zebrafish and in cultured cells, as the first step toward understanding genetic networks involved in the primitive heart tube formation (Specific Aim 2). Finally, from an ongoing zebrafish genetic screen, we identified a new mutation affecting the patterning of the primitive heart tube. We will characterize the phenotypes of this mutant and identify the molecular lesion causing this mutant phenotype (Aim 3). The combination of cellular, molecular and genetic studies proposed in this project will provide new and in-depth insight into mechanisms of primitive heart tube morphogenesis.