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精选鱼类能够在广泛的盐度范围（包括海水和淡水）中生活的分子机制。这些鱼被称为泛盐鱼，与大多数鱼类不同，它们具有适应机制，使它们能够忍受极端的盐度压力，并积极地抵消这种压力。本项目以罗非鱼为模型，旨在确定控制全盐鱼类盐度适应的分子信号通路。蛋白质组学方法（2D电泳、质谱、生物信息学）、组织微阵列/激光扫描细胞术分析、共免疫沉淀和重组DNA方法学将用于解决三个具体目标：OSTF1和14 - 3-3信号蛋白调控盐度适应的机制研究2. 盐度胁迫下由OSTF1和14-3-3介导的适应性触发信号（血浆皮质醇、血浆渗透压、介质盐度）的鉴定；3. 最近在PI实验室鉴定的15个新的盐度诱导罗非鱼基因的全长编码序列的确定和深入表征。这项工作的总体意义在于其固有的潜力，揭示了控制盐盐鱼类盐度适应和环境胁迫耐受的细胞信号转导的主要机制。了解这些机制对于理解为什么有些鱼类能很好地忍受环境压力，而另一些鱼类却对微小的环境变化很脆弱是至关重要的。通过研究环境对细胞和生物体功能的调节，科学知识的进步为预测、监测和抵消环境应激对生物体的负面影响提供了工具。因此，本项目的结果应该对一般细胞生物学的应激和环境适应具有广泛的适用性。罗非鱼是拟议研究的理想对象，因为该物种对美国和全世界的水产养殖都很重要。此外，罗非鱼具有很高的环境耐受性和适应性，是美国的入侵物种，从生态学的角度来看，罗非鱼是很重要的。该项目的重要影响还涉及博士后研究员和学生的深入培训，包括代表性不足的少数民族学生。此外，该项目还将支持各种研究、教育和推广活动。

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.