Basal body-mediated regulation of ciliary Ca2+ during cilia length control

 描述(由申请人提供)：本提案概述了为研究人员提供科学培训和职业发展技能的详细计划，以确保博士后培训期的成功。他的培训将由科罗拉多大学医学院的查德·皮尔森博士(赞助商)和李·尼斯万德博士(共同赞助商)指导。科学培训的重点将是第二信使离子钙(Ca~(2+))的水平如何在称为纤毛的亚细胞信号结构中调节，以及纤毛Ca~(2+)水平如何影响CIIA结构。纤毛是普遍存在的、进化上保守的附属物，几乎存在于人体的每一个细胞中。因为纤毛的缺陷造成了过多的破坏性 对于目前难治性发育障碍，这项研究有望提高我们对纤毛生物学的基本理解，并为未来的纤毛疾病治疗提供参考。为了研究纤毛与Ca~(2+)的关系，研究者提出假设，纤毛Ca~(2+)水平直接调节纤毛长度，而位于纤毛底部的基底体(BB)调节Ca~(2+)流入和流出CIIA。为了验证这一假设，研究人员提出了两个目标。在AIM1中，研究人员将使用钙离子成像、睫状肌钙离子的光学药理操作以及睫状肌钙离子水平的遗传消融来直接测试睫状肌钙离子水平如何影响纤毛长度。这一目标的结果将解决纤毛生物学中的一个基本问题，该问题与全局操纵钙离子如何能够特定地改变纤毛长度有关。在AIM2中，研究人员将使用类似的工具来定义结构缺陷的BBS如何改变钙流动 在静息和嗅觉感觉神经元的信号传递过程中，纤毛的进出。结果是 这些实验将阐明BB是一种新型的纤毛钙信号调节因子，这将为研究者开辟一个新的研究领域。调查员还与一支强大的指导团队一起确立了专业发展目标，例如编写手稿、参加国家会议和显微镜、数字图像分析和赠款撰写等领域的正式课程，以确保将科学培训转化为未来作为独立调查员的职业。

 DESCRIPTION (provided by applicant): This proposal outlines a detailed plan for providing the investigator with the scientific training and career developmen skills to ensure a successful postdoctoral training period. His training will be mentored by Drs. Chad Pearson (sponsor) and Lee Niswander (co-sponsor) at the University of Colorado-School of Medicine. The scientific training will focus on how levels of the second messenger ion calcium (Ca2+) are regulated within subcellular signaling structures called cilia, and, in turn, how cilia Ca2+ levels impact ciia structure. Cilia are ubiquitous and evolutionarily conserved appendages that are present on nearly every cell in the human body. Since defects in cilia contribute to a plethora of devastating and currently intractable developmental disorders, this research is expected to improve our basic understanding of cilia biology and inform future ciliopathy therapies. To investigate the relationship between cilia and Ca2+, the investigator has established the hypothesis that cilia Ca2+ levels directly regulate cilia length and that the basal body (BB), which is a small microtubule structure that sits at the base of cilia, regulates the flow of Ca2+ into and out of ciia. To test this this hypothesis, the investigator proposes to accomplish two aims. In Aim1, the investigator will use Ca2+ imaging, optopharmacological manipulation of ciliary Ca2+, and genetic ablation of ciliary Ca2+ levels to directly test how ciliary Ca2+ levels impact cilia lengt. The results from this aim will resolve a fundamental question in cilia biology that is related to how the global manipulation of Ca2+ is able to specifically alter cilia length. In Aim2, the investigator will use similar tools to define how structurally defective BBs alter the flow of Ca2+ into and out of cilia at rest and during signaling events in olfactory sensory neurons. The results from these experiments will illuminate the BB as a novel regulator of ciliary Ca2+ signaling, which will open up a new area of inquiry for the investigator to pursue. In conjunction with a strong mentoring team, the investigator has also established professional developmental goals, such as manuscript preparation, attendance at national conferences and formal coursework in the areas of microscopy, digital image analysis and grant writing to ensure that the scientific training is translated into a future career as an independent investigator.