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 正常生理需要感觉到渗透压变化和保持流体渗透性的能力 每个细胞的功能，因此对人类健康至关重要。流体失衡，由高渗透压引起 （高温性）或低渗透压（低渗透压），可能导致对器官的不可逆转损害 致命的神经创伤。在人类中，体液的渗透性在大脑中不断监测 保持在非常狭窄的范围内（275-299 MOSM/kg）。血液渗透压的增加一百分之一是 足以引发口渴。这种强大的渗透调节的关键要素是分子渗透传感器 检测渗透压变化。但是，动物界的渗透传感器的分子身份已经 仍然难以捉摸。在动物中识别渗透传感器的困难部分是由于缺乏公正 筛选系统。以前的识别动物渗透传感器的努力仅限于TRP通道。 但是，OSMOSENSOR不一定属于TRP渠道家族，因此可能已经避免了 检测。线虫C.秀丽隐杆线虫是研究渗透压的理想模型。像哺乳动物一样 已经鉴定出渗透传感系统和保守的信号分子。这是短暂的 生成时间（〜3天）以及易于且丰富的遗传工具，使秀丽隐杆线虫成为识别的理想系统 新颖的Osmosensors。为了识别秀丽隐杆线虫中的Osmosensor，我们设计并进行了神经活动 - 基于遗传筛查。我们已经将OSMS-1和OSMS-2确定为秀丽隐杆线虫中的候选OSMOSENSOR。 尽管有这一激动人心的发现，但许多问题仍未得到解决。在当前的建议中，我们建议测试 OSMS-1和OSMS-2是真正的OSMOSENSOR，并表征分子的假设 OSMS-1和OSMS-2感知渗透压的机制。我们将采用多学科的方法 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，我将接受有关钙成像，细胞培养，电生理记录和冷冻的广泛培训 Em。 K99/R00奖将使我能够在导师Drs的指导下获得这些技能。肖恩Xu 和梅兰妮（Melanie Ohi），这将帮助我启动独立的研究职业。拟议的工作将导致 鉴定动物王国中第一个渗透传感器，对 渗透压传感器感知渗透刺激，并提供有关渗透压，渗透调节和的新见解 相关的人类疾病。