Identification and Characterization of New Genes Required for Cilia Motility

描述（由申请人提供）：这项拟议研究的长期目标是进一步阐明原发性纤毛运动障碍（PCD）的机制。据估计，多达1：15000的个体患有PCD，其通常表现为慢性呼吸道感染、男性不育症、由于纤毛运动丧失或受损而导致的通常一定程度的原位逆转，以及在罕见情况下的囊性肾病。迄今为止，多种动力蛋白与PCD相关，然而，其他几个连锁的遗传基因座也与该疾病有关。在这些遗传间隔中与PCD发展相关的基因尚未被确定。目前，本实验室和其他实验室正在研究多个斑马鱼突变体，这些突变体在影响纤毛结构和功能的基因内具有病变。虽然大量的斑马鱼突变体表现出纤毛长度的缺陷，随后，他们的正常跳动的能力，只有少数拥有正常长度的纤毛，不能跳动。其中一个突变体是schmalhans，我们将用它来鉴定纤毛运动的重要基因。我们假设，斑马鱼瘫痪纤毛突变schmalhans窝藏在一个新的基因突变需要外动力蛋白臂组装。这些胚胎的特征是弯曲的体轴，鼻子，肾脏，椎管中的不动纤毛，并在受精后3天内发育肾囊肿。通过确定schmalhans的遗传病变，我们还希望发现和研究一个新的基因参与适当的肾功能的功能。我们的假设将通过以下具体目标进行验证：目的1.通过定位克隆和遗传间隔的计算机组装，吗啉代功能缺失研究和合成RNA的拯救来鉴定schmalhans基因。目标二。通过表达分析、与哺乳动物同源物的比较以及与PCD研究者的合作来表征schmalhans基因。目标3。通过鉴定相互作用蛋白和这些相互作用所需的Schmalhans结构域，确定Schmalhans影响纤毛运动的途径。最终，我们希望这项研究能够帮助其他人更好地识别和治疗PCD患者，使他们对潜在的疾病机制有更全面的了解。此外，这些研究可能有助于治疗与纤毛功能障碍有关的其他健康问题，如多囊肾病。

DESCRIPTION (provided by applicant): The long-term goal of this proposed research is to further elucidate the mechanisms underlying primary ciliary dyskinesia (PCD). As many as 1:15000 individuals are estimated to have PCD, which typically manifests as chronic respiratory tract infection, male infertility, often some degree of situs reversal due to lost or impaired cilia motility, and in rare cases, cystic kidney disease. To date, multiple dyneins have been associated with PCD, however, several other linked genetic loci have also been implicated in the disease. The genes relevant to development of PCD in these genetic intervals have yet to be identified. Multiple zebrafish mutants harboring lesions within genes that affect ciliary structure and function are currently being studied by this and other laboratories. While a large number of the zebrafish mutants exhibit defects in cilia length and subsequently, their ability to beat properly, only a few possess cilia of normal length that fail to beat. One such mutant is schmalhans, and we will use it to identify genes important for cilia motility. We hypothesize that the zebrafish paralyzed cilia mutant schmalhans harbors a mutation in a novel gene required outer dynein arm assembly. These embryos are characterized by a curved body axis, immotile cilia in the nose, kidneys, spinal canal, and develop kidney cysts within 3 days post-fertilization. By identifying the genetic lesion in schmalhans, we also hope to uncover and study the function of a new gene involved in proper kidney function. Our hypothesis will be tested by the following specific aims: Aim1.To identify the schmalhans gene through positional cloning and in silico assembly of the genetic interval, morpholino loss of function studies, and rescue using synthetic RNA. Aim 2. To characterize the schmalhans gene through expression analysis, comparison to mammalian homologs, and collaboration with PCD investigators. Aim 3. To identify the pathway(s) through which schmalhans acts to affect cilia motility by identifying interacting proteins and the domains of Schmalhans required for those interactions. Ultimately, it is our hope that this research will aid others to better identify and treat PCD patients by equipping them with a more complete understanding of the underlying disease mechanism. In addition, treatment of other health problems linked to cilia dysfunction, such as polycystic kidney disease, may be assisted by these studies.