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

基本信息

批准号：
10188127
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
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-03-01 至 2023-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10188127
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

项目摘要

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.

项目概要正常生理需要感知渗透压变化和维持液体渗透压的能力每个细胞的功能，因此对人类健康至关重要。高渗透压引起的体液失衡（高渗透压）或低渗透压（低渗透压），可导致器官不可逆转的损害并导致致命的神经创伤。在人类中，体液的渗透压在大脑中被持续监测，并且保持在非常窄的范围内（275-299 mOsm/kg）。血液渗透压仅增加百分之一足以引起口渴。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。然而，动物界渗透传感器的分子特性已经仍然难以捉摸。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。先前识别动物渗透传感器的努力仅限于 TRP 通道。然而，渗透传感器不一定属于 TRP 通道家族，因此可能已被忽视检测。线虫秀丽隐杆线虫是研究渗透传感的理想模型。与哺乳动物一样，秀丽隐杆线虫渗透传感系统和保守的信号分子已被鉴定。这，连同它的短世代时间（约 3 天）和简便且丰富的遗传工具，使秀丽隐杆线虫成为鉴定的理想系统新型渗透传感器。为了识别秀丽隐杆线虫中的渗透传感器，我们设计并进行了神经活动 - 基于基因筛选。我们已确定 OSMS-1 和 OSMS-2 为线虫的候选渗透传感器。尽管有这一令人兴奋的发现，但许多问题仍未得到解答。在当前的提案中，我们建议测试 OSMS-1 和 OSMS-2 是真正的渗透传感器并表征分子的假设 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、电生理学和冷冻电镜。要做的事因此，我将接受钙成像、细胞培养、电生理记录和冷冻方面的广泛培训 EM。 K99/R00 奖将使我能够在导师 Drs. 的指导下获得这些技能。徐肖恩和 Melanie Ohi，这将帮助我开展独立的研究生涯。拟议的工作将导致鉴定动物王国中第一个渗透传感器，从分子角度了解如何渗透传感器感知渗透刺激，并为渗透感觉、渗透调节和渗透提供新的见解。相关人类疾病。