Kurly's Role in Left-Right Patterning and Congenital Heart Disease

描述（由申请人提供）：脊椎动物具有沿左右轴的内部不对称，包括器官的不对称放置。器官（如心脏）的正确放置对胚胎的正常发育和存活至关重要。由于基因表达不当，每5000个孩子中就有1个天生有左右模式缺陷，通常表现为先天性心脏病。了解在胚胎中建立正确的左右模式所涉及的遗传途径对于理解这些缺陷是如何产生的至关重要。一种新的斑马鱼突变体，Kurly，影响左右模式和随后的心脏发育。有趣的是，Kurly的新人类同源物映射到位于唐氏综合症关键区域的人类21号染色体上的一个基因。位于该区域的基因可能是唐氏综合症患者先天性心脏缺陷高发的原因。一般人群中有0.8%患有先天性心脏缺陷，而患有唐氏综合症的人中有40-50%患有先天性心脏缺陷。通过了解Kurly如何在斑马鱼中发挥作用，将获得有价值的见解，了解左右模式缺陷和随后的先天性心脏缺陷是如何产生的。斑马鱼是一种优秀的模式生物，因为它的外部发育，透明度和基因操作可以进行。此外，由于许多参与左右模式的基因在人类中具有保守的作用，因此在斑马鱼身上进行的实验和从这些实验中获得的知识显然适用于更大的社会利益。为了研究Kurly对左右脑模式和心脏发育的影响，我将采取多种方法。我将首先使用免疫组织化学和荧光蛋白标记来检查Kurly的功能位置和部位。然后，我将通过注射测定和免疫荧光来探讨在这些早期图案事件中，Kurly如何影响初级纤毛和细胞极性。在这些发育阶段，我将通过串联亲和纯化最终确定与Kurly相互作用的其他蛋白质，以便深入了解可能需要Kurly功能的重要途径。相关性：这项研究通过对一种基因的深入分析，极大地造福了人类，这种基因在调控不当时，会导致左右内脏器官模式缺陷和先天性心脏缺陷。

DESCRIPTION (provided by applicant): Vertebrates have internal asymmetries along the left-right axis, including asymmetric placement of the organs. Correct placement of organs, such as the heart, is critical for proper embryonic development and survival. Due to improper gene expression as many as one in 5,000 children are born with left-right patterning defects which often manifest as congenital heart disease. Understanding the genetic pathways involved in establishing proper left-right patterning in the embryo is vital to understanding how these defects may arise. A novel zebrafish mutant, Kurly, affects left-right patterning and subsequent heart development. Interestingly, the novel human ortholog of Kurly maps to a gene located on human chromosome 21 within the Down Syndrome Critical Region. Genes located in this area are perhaps responsible for the high incidence of congenital heart defects in Down Syndrome patients. While 0.8% of the general population is affected by congenital heart defects, 40-50% of those affected by Down Syndrome have congenital heart defects. By understanding how Kurly functions in zebrafish, valuable insight will be gained into how left-right patterning defects and subsequent congenital heart defects may arise. The zebrafish is an excellent model organism due to its external development, transparency, and genetic manipulations that can be made. Also, as many genes involved in left-right patterning have conserved roles in humans, experiments performed in zebrafish and knowledge gained from these experiments are clearly applicable to the greater good of society. To investigate Kurly's effects on left- right patterning and heart development, I will take multiple approaches. I will first use immunohistochemistry and fluorescent protein tagging to examine the location and sites of function for Kurly. I will then explore how Kurly affects primary cilia and cell polarity during these early patterning events by injection assays and immunofluorescence. I will finally identify other proteins that interact with Kurly, through tandem affinity purification, during these stages of development in order to gain insight into significant pathways which may require Kurly function. Relevance: This research greatly benefits mankind through an in-depth analysis of a gene that, when misregulated, leads to left-right internal organ patterning defects as well as congenital heart defects.