Determining the roles of Ca2+-binding proteins in modulation of hair cell mechanotransduction

摘要 首席研究员在休斯顿德克萨斯大学健康科学中心获得博士学位 他所做的工作是为了了解脊髓损伤引起的慢性疼痛的潜在机制 埃德加·T·沃尔特斯 (Edgar T. Walters) 博士的监督。然后，他搬到拉霍亚的斯克里普斯研究所 (TSRI)，在那里 他加入了 Ulrich Mueller 博士的实验室，接受听觉感知训练，特别是机械传导方面的训练。 耳蜗毛细胞。最近，他与穆勒博士一起搬到了约翰·霍普金斯大学 (JHU)，那里有许多顶尖大学 感觉生物学领域的科学家可以立即联系到他，并在那里继续他的博士后研究金 获得分子生物学和小鼠建模方面的培训。此外，协助候选人进行具体的 技术，帮助他确定工作机会，为工作面试做好准备，并帮助他建立一个实验室，一个实验室 顾问小组成立。该团队包括博士。乌尔里希·穆勒 (Ulrich Mueller)、伊丽莎白·格洛瓦茨基 (Elisabeth Glowatzki)、吴斌 (Bin Wu) 和赵祝 Qiu，JHU 的研究人员，拥有有关职业发展和实验室的最新知识 管理。最后，拟议的课程、科学会议和研讨会将补充他的培训 计划来实现他成为一名成功的独立调查员的目标。 拟议工作的长期研究目标是寻求该领域具有挑战性的问题的答案 感觉生物学 1) 毛细胞中的机械传导通道的组成部分是什么，以及 2) 负责调节毛细胞机械转导的机制是什么。为了实现这些目标， 他将特别关注两类蛋白质：(i) 在酵母双杂交筛选中鉴定出的蛋白质 与 TMHS/LHFPL5 相互作用，TMHS/LHFPL5 是毛细胞机械转导复合体的一个组成部分； (ii) Ca2 结合 作为调节转导的候选蛋白质。研究计划提出检验中心假设 LHFPL5 和与其结合的蛋白质是毛细胞中更大的机械转导通道复合体的一部分。 一些蛋白质可能对通道孔有贡献，而另一些蛋白质可能发挥调节作用。钙离子 预计结合蛋白对于转导机制的有据可查的调节至关重要 至 Ca2 。拟议的研究将分析这些蛋白质在耳蜗机械转导中的作用 毛细胞和前庭毛细胞。将使用以下实验策略来测试蛋白质功能： 电生理学、生物化学和小鼠建模。研究结果将为支持该项目提供坚实的基础 首席研究员的研究目标有望提供 1) 更好地理解力转导 毛细胞中的通道，2）深入了解通道调节机制（例如适应）。

ABSTRACT The principal investigator received his PhD at the University of Texas Health Science Center at Houston for the work he performed to understand the mechanisms underlying spinal cord injury-induced chronic pain under the supervision of Dr. Edgar T. Walters. He then moved to The Scripps Research Institute (TSRI), La Jolla, where he joined Dr. Ulrich Mueller's laboratory to train in auditory perception, specifically in mechanotransduction of cochlear hair cells. Recently, he moved with Dr. Mueller to Johns Hopkins University (JHU), where many top scientists in sensory biology are immediately available and where he continues his postdoctoral fellowship to obtain training in molecular biology and mouse modeling. Furthermore, to assist the candidate with specific techniques, help identify job opportunities, prepare him for job interviews, and help him build a laboratory, an advisory team was formed. This team includes Drs. Ulrich Mueller, Elisabeth Glowatzki, Bin Wu and Zhaozhu Qiu, who are investigators at JHU with the most current knowledge about career development and lab management. Finally, proposed courses, scientific meetings and seminars will complement his training program to accomplish his goal of becoming a successful independent investigator. The long-term research goal of the proposed work is to seek answers to challenging questions in the area of sensory biology as to 1) what are the components of the mechanotransduction channel in hair cells, and 2) what is the mechanism responsible for modulation of hair cell mechanotransduction. To achieve these goals, he will specifically focus on two classes of proteins: (i) proteins identified in a yeast two-hybrid screen that interact with TMHS/LHFPL5, a component of the mechanotransduction complex of hair cells; (ii) Ca2+ binding proteins that are candidates to regulate transduction. The research plan proposes to test the central hypothesis that LHFPL5 and proteins that bind to it are part of a larger mechanotransduction channel complex in hair cells. Some of the proteins might contribute to the channel pore while others might play modulatory roles. Ca2+ bindings proteins are predicted to be critical for the well-documented regulation of the transduction machinery to Ca2+. The proposed studies will analyze the role of these proteins in mechanotransduction in both cochlear hair cells and vestibular hair cells. Protein function will be tested using an experimental strategy that combines electrophysiology, biochemistry, and mouse modeling. The results will provide a solid foundation to support the principal investigator's research goals that promise to offer 1) a better understanding of mechanotransduction channels in hair cells, and 2) insights into the mechanisms of channel regulation such as adaptation.