MCA: Cellular Responses to Thermal Stress in Antarctic Fishes: Dynamic Re-structuring of the Proteome in Extreme Stenotherms

MCA：南极鱼类对热应激的细胞反应：极端钝温鱼蛋白质组的动态重组

• 批准号: 2322117
• 负责人: Bradley Buckley
• 金额: $ 30.01万
• 依托单位: Portland State University
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2024
• 资助国家: 美国
• 起止时间: 2024-03-01 至 2027-02-28
• 项目状态: 未结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2322117&HistoricalAwards=false
• 关键词: MCA; Cellular; Responses; Thermal; Stress

Part 1: This project focuses on a group of ecologically important species of fishes which inhabit the frigid waters of Antarctica. They represent a key link in the polar food web as they are prey for penguins, seals and toothed whales. These fish have evolved in the constant, extreme cold for millions of years and therefore, are very sensitive to the increasing water temperatures associated with global warming. These studies will investigate the impacts of incremental heat exposure on the biology of these fishes by examining their ability to respond, or inability to respond, to elevated temperatures. The project will employ cutting-edge technology to examine responses at the cellular level that may help these environmentally sensitive fishes adapt to the challenges of global warming. The primary goal is to increase our collective understanding of how polar ecosystems are likely to be impacted in the coming decades.Part 2: The proposed research is designed to use an existing bank of frozen tissues from a species of cold-adapted Antarctic fish to investigate protein-level responses to heat stress. These samples were collected earlier in the PI's career during fieldwork at McMurdo Station, Antarctica. Four tissues (control as well as heat- stressed) will be analyzed via mass spectrometry to characterize their proteome, defined as the entire complement of proteins in a sample. This includes both identification and quantification of these proteins. The goal is to determine what mechanisms of response to elevated temperature are available to the extremely cold-adapted, stenothermic fishes of Antarctica. Follow-up analyses will use immunoblotting (Western blotting) with antibodies specific to a sub-set of proteins revealed to be heat-responsive in the proteomic analyses. As this is a Mid-Career Advancement Award, training and mentorship in proteomic analyses for the PI will be supported, with time spent at the partner institution, the University of California, Davis. Intellectual MeritWhile there has been an increase in the use of genomic technologies to probe gene expression profiles in Antarctic species, few studies exist looking at protein level changes during exposure to heat stress in these organisms. Therefore, the proposed studies would represent a large leap forward in our understanding of how these environmentally sensitive species can, or cannot, respond at the cellular level as the Earth continues to warm and water temperatures rise. As proteins do the "work" in the cell, it's vital to understand which proteins are present and in what quantity and how dynamic this "proteome" is during stress. The proposed studies would provide this information for thousands of proteins, using already existing samples. The findings would be entirely novel and would allow us a much better picture of how animals that evolved in the cold for millions of years are likely to respond to climate change. Broader ImpactsThe PI has established relationships with several regional K-12 institutions and will continue to provide outreach in the form of classroom visits and the creation of classroom curricula. The PI has an on-going collaboration with the Oregon Coast Aquarium (Newport, OR) to create novel teaching materials for grades 6-8. The Aquarium has partners in surrounding school districts and will help disseminate videos about marine biology and climate change. Modules concerning polar species will be created under this proposal. An interactive website will be created demonstrating the Antarctic food web. All of the proteomic analyses and libraries generated under this award will be made publicly available for use by any interested researcher.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

第1部分：该项目着重于一群居住在南极洲寒冷的水域的生态重要的鱼类。它们代表了极地食品网中的关键环节，因为它们是企鹅，海豹和齿鲸的猎物。这些鱼类在数百万年内的恒定寒冷中进化，因此对与全球变暖相关的水温升高非常敏感。这些研究将通过检查它们对温度升高的反应或无法做出反应的能力来研究增量热暴露对这些鱼类生物学的影响。该项目将采用尖端技术来检查细胞水平的反应，这可能有助于这些对环境敏感的鱼类适应全球变暖的挑战。主要目标是提高我们对未来几十年中极性生态系统可能受到影响的集体理解。第2部分：拟议的研究旨在利用现有的冷冻组织库，这些组织来自一种冷适应的南极鱼类，以研究蛋白质水平的热应力反应。这些样品是在南极麦克默多车站的野外工作期间在PI的职业生涯中早些时候收集的。将通过质谱法分析四个组织（对照和热应力），以表征其蛋白质组，定义为样品中蛋白质的整个补体。这包括对这些蛋白质的识别和定量。目的是确定对温度升高的反应机制，可用于极度冷适应的南极洲的炎热鱼类。后续分析将使用针对蛋白质子群特异性的抗体的免疫印迹（蛋白质印迹），该蛋白在蛋白质组学分析中揭示具有热反应性。由于这是一项职业发展奖，因此将支持PI蛋白质组学分析的培训和指导，并在加利福尼亚大学戴维斯分校的合作伙伴机构花费的时间。智力优点是，基因组技术在南极物种中使用基因组技术的使用有所增加，但在暴露于这些生物体热应激期间蛋白质水平的变化几乎没有研究。因此，拟议的研究将代表我们对这些对环境敏感的物种如何或无法在地球持续温暖和水温升高时在细胞水平上做出反应的巨大飞跃。正如蛋白质在细胞中进行“工作”，了解哪些蛋白质存在以及在压力期间这种“蛋白质组”的动态多么动态至关重要。拟议的研究将使用已经存在的样品为数千种蛋白质提供此信息。这些发现将完全是新颖的，可以更好地了解数百万年的寒冷动物如何对气候变化做出反应。 PI更广泛的影响与多个区域K-12机构建立了关系，并将继续以课堂访问的形式提供外展和教室课程的创建。 PI与俄勒冈海岸水族馆（OR）进行了持续的合作，为6 - 8年级创建了新颖的教材。该水族馆有周围学区的合作伙伴，将有助于传播有关海洋生物学和气候变化的视频。有关极性物种的模块将根据该提案创建。将创建一个互动网站，展示南极食品网。任何感兴趣的研究人员都将公开使用根据该奖项生成的所有蛋白质组学分析和图书馆。该奖项反映了NSF的法定任务，并认为使用基金会的知识分子优点和更广泛的影响审查标准，认为值得通过评估值得支持。