Osmosensory signal transduction in gill cells of euryhaline tilapia

该项目的总体目标是研究使包括罗非鱼在内的特定鱼类群体能够在包括海水和淡水在内的广泛盐度环境中生活的分子机制。这些鱼被称为泛盐目鱼类，与大多数鱼类不同，它们拥有适应机制，使它们能够忍受极端的盐度压力，并积极地抵消这种压力。以罗非鱼为模型，本项目旨在确定控制泛盐度鱼类盐度适应的分子信号通路。蛋白质组学方法(二维电泳、质谱学、生物信息学)、组织芯片/激光扫描细胞学分析、免疫共沉淀和重组DNA方法将针对三个特定目标：1.研究OSTF1和14 3-3信号蛋白控制盐度适应的机制；2.识别在盐胁迫过程中触发OSTF1和14-3-3介导的适应的信号(血浆皮质醇、血浆渗透压、中等盐度)；3.测定全长编码序列并对最近在PI实验室发现的15个新的盐度诱导罗非鱼基因进行深入分析。这项工作的总体意义在于其内在的潜力，揭示了控制泛盐目鱼类盐度适应和环境胁迫耐受的主要细胞信号转导机制。对这些机制的了解对于理解为什么一些鱼类很好地耐受环境压力，而其他物种即使是微小的环境变化也很容易受到影响，是至关重要的。通过研究细胞和生物体功能的环境调节来促进科学知识的进步，有利于为预测、监测和抵消环境应激对生物体的负面影响提供工具。因此，该项目的结果应该对应激和环境适应的一般细胞生物学具有广泛的适用性。罗非鱼是建议研究的理想物种，因为这种物种对美国和世界各地的水产养殖都很重要。此外，从生态学的角度来看，罗非鱼很重要，因为它们是美国的入侵物种，因为它们具有很高的环境压力耐受性和适应性。该项目的重要影响还涉及深入培训博士后研究人员和学生，包括代表人数不足的少数族裔学生。此外，该项目还将支持各种研究、教育和外展活动。

The overall objective of this project is to investigate molecular mechanisms that enable a select group of fishes, including tilapia, to live in a wide range of salinity, including seawater and fresh water. These fishes are called euryhaline and, unlike most fishes, they possess adaptive mechanisms that allow them to tolerate extreme salinity stress and to actively counteract such stress. Using tilapia as a model this project aims to determine the molecular signaling pathways that control salinity adaptation in euryhaline fishes. Proteomics methods (2D electrophoresis, mass spectrometry, bioinformatics), tissue microarray/ laser scanning cytometry analysis, co-immunoprecipitation, and recombinant DNA methodology will be used to address three specific aims: 1. Investigation of the mechanisms by which OSTF1 and 14 3-3 signaling proteins control salinity adaptation; 2. Identification of the signal (plasma cortisol, plasma osmolality, medium salinity) that triggers adaptations mediated by OSTF1 and 14-3-3 during salinity stress; 3. Determination of full-length coding sequence and in-depth characterization of 15 novel salinity-induced tilapia genes recently identified in the PI's laboratory. The overall significance of this work lies in its inherent potential for unraveling major mechanisms of cellular signal transduction that control salinity adaptation and environmental stress tolerance in euryhaline fishes. Knowledge about these mechanisms is critical for understanding why some fish species tolerate environmental stress very well whereas other species are vulnerable to even minor environmental change. The advancement of scientific knowledge by the study of environmental regulation of cell and organismal function has the benefit of providing the tools for predicting, monitoring, and counteracting negative effects of environmental stress on organisms. Thus, the results of this project should have broad applicability to the general cell biology of stress and environmental adaptation. Tilapia are ideal for the proposed studies because this species is important for aquaculture in the US and worldwide. Moreover, tilapia are important from an ecological perspective as they are an invasive species in the US because of their high environmental stress tolerance and adaptability. Important broader impacts of this project also pertain to in-depth training of a postdoctoral researcher and students, including underrepresented minority students. Furthermore, a variety of research, educational, and outreach activities will be supported by this project.