Identification of novel osmosensing receptors in C. elegans

秀丽隐杆线虫中新型渗透感应受体的鉴定

• 批准号: 10360684
• 负责人: Xinxing Zhang
• 金额: $ 10万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-03-01 至 2023-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10360684
• 关键词: Address; Animals; Award; Behavioral; Biochemistry; Blood; Body Fluids; Brain; Caenorhabditis elegans; Calcium; Categories; Cell Culture Techniques; Cells; Chemicals; Cryoelectron Microscopy; Detection; Disease; Electrophysiology (science); Elements; Family; G-Protein-Coupled Receptors; Generations; Genetic; Genetic Screening; Health; Hearing; Homeostasis; Homologous Gene; Human; Image; In Vitro; Knowledge; Lead; Life; Liquid substance; Mammalian Cell; Mammals; Mediating; Mentors; Modality; Modeling; Molecular; Molecular Genetics; Monitor; Nature; Nematoda; Nervous System Trauma; Neurons; Organ; Organism; Osmolar Concentration; Osmoregulation; Phase; Photons; Physiological; Property; Proprioception; Regulation; Research; Research Personnel; Rodent; Sensory; Sensory Receptors; Signaling Molecule; Smell Perception; Stimulus; Structure; System; TRP channel; Taste Perception; Temperature; Testing; Thirst; Time; Touch sensation; Training; Vision; Work; avoidance behavior; base; career; design; drinking; falls; human disease; in vivo; insight; interdisciplinary approach; mechanical force; mutant; novel; post-doctoral training; receptor; relating to nervous system; screening; sensory stimulus; skills; tool

Project Summary The ability to sense osmolarity changes and maintain fluid osmolarity is required for normal physiological functions of every single cell and thus vital for human health. Fluid imbalance, caused by high osmolarity (hyperosmolarity) or low osmolarity (hypoosmolarity), can lead to irreversible damage to organs and cause lethal neurological trauma. In humans, the osmolarity of body fluids is continuously monitored in the brain and kept within a very narrow range (275-299 mOsm/kg). As little as a one percent increase in blood osmolarity is enough to trigger thirst. A key element for such robust osmoregulation is the molecular osmosensors that detect osmolarity changes. However, the molecular identities of osmosensors in the animal kingdom have remained elusive. The difficulty in identifying osmosensors in animals is partly due to the lack of an unbiased screening system. Previous efforts to identify osmosensors in animals were restricted to TRP channels. However, osmosensors do not necessarily fall into the TRP channel family and thus may have eluded detection. The nematode C. elegans is an ideal model to study osmosensing. Like mammals, C. elegans has osmosensing systems, and conserved signaling molecules have been identified. This, together with its short generation time (~3 days) and facile and rich genetic tools, makes C. elegans an ideal system for identifying novel osmosensors. To identify osmosensors in C. elegans, we designed and conducted a neural activity- based genetic screen. We have identified OSMS-1 and OSMS-2 as candidate osmosensors in C. elegans. Despite this exciting finding, many questions remain unanswered. In the current proposal, we propose to test the hypothesis that OSMS-1 and OSMS-2 are bona fide osmosensors and characterize the molecular mechanisms by which OSMS-1 and OSMS-2 sense osmolarity. We will take a multidisciplinary approach by integrating molecular genetics, behavioral analysis, calcium imaging, electrophysiology, and cryo-EM. To do so, I will receive extensive training on calcium imaging, cell culture, electrophysiological recording, and cryo- EM. The K99/R00 award will allow me to acquire these skills with guidance from my mentors Drs. Shawn Xu and Melanie Ohi, which will help me to launch an independent research career. The proposed work will lead to the identification of the first osmosensors in the animal kingdom, gain a molecular understanding of how osmosensors sense osmotic stimuli, and provide novel insights into osmosensation, osmoregulation and related human diseases.

项目摘要 感知渗透压变化和维持液体渗透压的能力是正常生理功能所必需的。 每一个细胞的功能，因此对人类健康至关重要。液体失衡，由高渗透压引起 渗透压（高渗透压）或低渗透压（低渗透压）可导致器官不可逆损伤， 致命的神经创伤在人类中，体液的渗透压在大脑中被持续监测， 保持在非常窄的范围内（275-299 mOsm/kg）。血液渗透压增加1%， 足以引起口渴。这种强大的生物调节的关键因素是分子生物传感器， 检测渗透压变化。然而，在动物王国中，生物传感器的分子特性 仍然难以捉摸识别动物渗透传感器的困难部分是由于缺乏公正的方法 筛选系统以前在动物中识别TRP传感器的努力仅限于TRP通道。 然而，MEMS传感器不一定属于TRP通道族，因此可能已经避开了TRP通道。 侦测线虫C. elegans是一种理想的生物传感模型。与哺乳动物一样，C. elegans有 已经鉴定了生物传感系统和保守的信号分子。这一点，连同其短 世代时间（~ 3d）和简便而丰富的遗传工具，使C.一个理想的系统， 新颖的MEMS传感器。识别C中的传感器。我们设计并进行了一种神经活动 基因筛查我们已经确定OSMS-1和OSMS-2为C.优雅的 尽管这一令人兴奋的发现，许多问题仍然没有答案。在目前的提案中，我们建议测试 假设OSMS-1和OSMS-2是真正的生物传感器，并表征了分子 OSMS-1和OSMS-2感知渗透压的机制。我们将采取多学科方法， 整合分子遗传学、行为分析、钙成像、电生理学和冷冻EM。做 因此，我将接受广泛的培训，包括钙成像、细胞培养、电生理记录和冷冻- EM. K99/R 00奖将让我在导师Shawn Xu博士的指导下获得这些技能 和梅勒妮·奥希，这将帮助我开始一个独立的研究生涯。拟议的工作将导致 在动物王国中识别出第一个生物传感器， 渗透压传感器感知渗透压刺激，并为渗透压感觉、渗透压调节和渗透压调节提供了新的见解。 相关的人类疾病。