RUI: Hydra TRP Channels: Expression and Functional Analysis

RUI：Hydra TRP 通道：表达和功能分析

• 批准号: 1243855
• 负责人: Susan McLaughlin
• 金额: $ 18.26万
• 依托单位: CUNY Queensborough Community College
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-03-01 至 2018-02-28
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1243855&HistoricalAwards=false
• 关键词: RUI; Hydra; TRP; Channels; Expression; RUI; Hydra; TRP; Channels; Expression

INTELLECTUAL MERIT Transient receptor potential (TRP) channels are a superfamily of cation channels that regulate processes as diverse as olfaction, thermal perception, pain, synaptic transmission and development. They can be activated by both environmental stimuli (e.g. chemicals, light, temperature, pressure) and intracellular stimuli (e.g. calcium and G protein coupled receptors). More than 50 different TRP channels have been identified in organisms ranging from yeasts and protists to mammals. The experiments in this project will extend the study of TRP channels into one of the most primitive metazoans with a nervous system, the cnidarian Hydra. A characteristic of all the Cnidaria is the possession of stinging cells called cnidocytes, which in Hydra are responsible for prey capture, adhesion and locomotion. Although significant progress has been made in understanding development and regeneration in Hydra, the molecular mechanisms that control Hydra behaviors such as its feeding response and cnidocyte release are as of yet poorly understood. An examination of the Hydra genome revealed the presence of multiple candidate Hydra TRP channel genes. Whole mount in situ hybridization will be used to localize TRP channel expression to specific Hydra body regions. TRP channel function will be examined by testing known chemical TRP channel activators and inhibitors to determine if they affect Hydra feeding behavior or cnidocyte release, and if they affect the ability of Hydra to regenerate or to form buds during asexual reproduction. In addition, RNA interference will be used to silence the expression of specific Hydra TRP channels. The resulting TRP(-) hydra will then be tested in the functional assays to determine the effect of TRP channel gene silencing. The study of Hydra TRP channels could answer fundamental questions about the signal transduction systems involved in Hydra behaviors and development. A comparison of the primitive TRP channels in Hydra with the more recently evolved TRP channels of higher organisms will provide a useful mechanism for both structure-function analysis and for studying evolutionary relationships. BROADER IMPACTS Queensborough Community College (QCC) is an urban community college with a student population comprised of nearly equal numbers of Asians, Caucasians, Latinos and African Americans. Many of QCC's students come from low-income families, and are the first in their family to attend college. One of the goals of this project is to afford QCC undergraduates the opportunity to do research in a scientific laboratory. In the course of their research, students will learn Hydra culture methods, basic molecular biology techniques and DNA/protein sequence analysis. As the research in this project spans the spectrum from DNA sequence to development and behavior, students will learn how to make connections between events at the molecular level and those occurring at the level of the entire organism. In addition, since the TRP channel gene family is found in both very simple organisms such as yeast and in complex organisms like mammals, students will need to think in evolutionary terms when analyzing TRP channel function. Firsthand experience gained by working in a laboratory allows students to connect the theoretical concepts they learn in the classroom with real-world experiences. This will enhance their understanding of scientific concepts, and will give them the opportunity to apply their knowledge to their experiments in the laboratory. Student participation in research increases retention and graduation rates, a particularly important benefit at a community college like QCC, and undergraduates who are exposed to research early during their college careers are more likely to choose STEM majors and to remain in STEM careers.

智力功绩瞬态受体电位（TRP）通道是阳离子通道的超家族，它们调节了嗅觉，热感知，疼痛，突触传播和发育等多样化的过程。它们可以通过环境刺激（例如化学物质，光，温度，压力）和细胞内刺激（例如钙和G蛋白偶联受体）激活它们。从酵母和生物到哺乳动物的生物中已经确定了50多种不同的TRP通道。该项目中的实验将将TRP通道的研究扩展到具有神经系统Cnidarian Hydra的最原始的后代之一。所有cnidaria的一个特征是拥有称为cnidocytes的细胞，在Hydra中，该细胞负责捕食，粘附和运动。尽管在理解Hydra的发展和再生方面取得了重大进展，但控制Hydra行为（例如其进食反应和CNIDOCYTE释放）的分子机制尚未了解。对Hydra基因组的检查表明存在多个候选Hydra TRP通道基因。整个原位杂交将用于将TRP通道表达式定位到特定的Hydra身体区域。 TRP通道功能将通过测试已知的化学TRP通道激活剂和抑制剂来确定它们是否影响九头蛇的进食行为或cnidocyte释放，以及它们是否影响九头士在无性繁殖过程中的再生能力或形成芽的能力。另外，RNA干扰将用于沉默特定的Hydra TRP通道的表达。然后，将在功能测定中测试所得的TRP（ - ）HYDRA，以确定TRP通道基因沉默的效果。 Hydra TRP通道的研究可以回答有关与九头蛇行为和发展有关的信号转导系统的基本问题。将Hydra的原始TRP通道与较最近进化的高生物体进化的TRP通道进行比较将为结构功能分析和研究进化关系提供有用的机制。昆斯伯勒社区学院（QCC）的更广泛影响是一所城市社区学院，其学生人数几乎由亚洲人，高加索人，拉丁裔和非裔美国人组成。 QCC的许多学生来自低收入家庭，是他们家庭中第一个上大学的人。该项目的目标之一是为QCC提供了在科学实验室进行研究的机会。在研究过程中，学生将学习Hydra培养方法，基本分子生物学技术和DNA/蛋白质序列分析。随着该项目的研究跨越了从DNA序列到发展和行为的频谱，学生将学习如何在分子级别与发生在整个有机体水平上发生的事件之间建立联系。此外，由于在酵母等非常简单的生物和哺乳动物等复杂生物中都发现了TRP通道基因家族，因此在分析TRP通道功能时，学生将需要以进化的方式进行思考。通过在实验室工作获得的第一手经验使学生可以将他们在课堂上学习的理论概念与现实世界的经验联系起来。这将增强他们对科学概念的理解，并将使他们有机会将知识应用于实验室的实验。学生参与研究会提高保留率和毕业率，这是QCC等社区大学的特别重要好处，以及在大学职业生涯中早期接受研究的本科生更有可能选择STEM专业并留在STEM职业中。