Investigating the processes of life in the cold: high resolution imaging of cellular proteostasis in Antarctic fish species

研究寒冷中的生命过程：南极鱼类细胞蛋白质稳态的高分辨率成像

• 批准号: 2408885
• 金额: --
• 依托单位: University of Cambridge
• 依托单位国家: 英国
• 项目类别: Studentship
• 财政年份: 2020
• 资助国家: 英国
• 起止时间: 2020 至 无数据
• 项目状态: 未结题
• 来源: https://gtr.ukri.org/projects?ref=studentship-2408885
• 关键词: Investigating; processes; life; cold; resolution

Antarctic ecosystems have evolved in a singularly isolated environment for millions of years. Due to a combination of geological and climatic factors, the Antarctic is the most isolated ecosystem on earth. This shielded environment has been stable for over 10 million years, giving rise to a remarkable diversity in adaptation to this continent. Antarctica has the highest proportion of endemic species of all ecosystems with around 17,000 marine invertebrate species. [1] Antarctic climate is however now facing an unprecedented threat. The Intergovernmental Panel on Climate Change's Five reasons for Concern [2] report the poles as being the fastest warming climates. The combined effects of an ecosystem that has been stable for millions of years and its projected rapidly changing climate make Antarctic cold-adapted species especially vulnerable to ocean climate change. The vulnerability of these species lies in the stability of their cellular machinery, and in particular the making, folding and maintaining of proteins. Very little is currently known about the mechanisms of protein stability around 0 C, and fewer still about this stability in complex organisms such as Antarctic fish. In order to understand the fundamental mechanisms of life in the cold, and to apply this understanding to incoming changes in Antarctic ecosystems, the British Antarctic Survey has partnered with several groups of the University of Cambridge and others in the UK in an ambitious and highly interdisciplinary project. As part of this collaboration, this PhD project is specifically aiming at the development of microscopy and fluorescence tools to study the protein lifecycle of Antarctic species. This PhD project will be focused on the development of a microscopy platform (year 1) along with fluorescence reporter methods (year 2). Indeed, microscopy at cold temperatures comes with technical challenges. These include the appropriate dosage of laser light to maintain physiological conditions in the cells, and a tight control of the microscope stage temperature so as to observe the samples across a range of conditions. We also expect challenges linked with the physics of the microscopy materials themselves (contraction of different parts, change in refractive indices... ). Once the microscope itself is adapted to polar conditions, we will also have to develop fluorescence tools operational around 0 C. These tools will be used to study factors affecting protein folding and stability, such as viscosity and cell chemistry. Two methods to measure intracellular viscosities will be tried, namely Single Particle Tracking (looking at the movement of one element over time) and Molecular Rotors (which rotate more or less quickly depending on cell conditions). We will use the combination of the microscopy and fluorescence techniques in the cold to link cellular phenotypes with the change of temperature experienced by the cell (year 3). This project is highly interdisciplinary and exists at the intersection of Biology, Physics and Engineering, bringing together a wide variety of scientific techniques. In terms of the EPSRC's strategies and research areas, this project touches on the following: Analytical sciences through the development of a new and unique microscopy platform, Biophysics and soft matter physics through the study of fundamental protein folding dynamics, or Sensors and Instrumentation through the development of protein tracking and temperature and viscosity measurements in

几百万年来，南极生态系统一直在一个异常孤立的环境中进化。由于地质和气候因素的综合作用，南极是地球上最孤立的生态系统。这种被屏蔽的环境已经稳定了1000多万年，导致了适应这块大陆的显著多样性。在所有生态系统中，南极洲特有物种的比例最高，约有17,000种海洋无脊椎动物。[1]然而，南极气候现在正面临前所未有的威胁。政府间气候变化专门委员会的五个令人担忧的原因[2]报告称，两极是变暖最快的气候。数百万年来一直稳定的生态系统及其预计迅速变化的气候的综合影响，使南极适应寒冷的物种特别容易受到海洋气候变化的影响。这些物种的脆弱性在于其细胞结构的稳定性，特别是蛋白质的制造、折叠和维持。目前，人们对蛋白质在0℃附近的稳定性机制知之甚少，对南极鱼类等复杂生物的这种稳定性更是知之甚少。为了了解寒冷中生命的基本机制，并将这一理解应用于南极生态系统即将发生的变化，英国南极调查局与剑桥大学和英国其他几个小组合作开展了一个雄心勃勃的、高度跨学科的项目。作为这项合作的一部分，这个博士项目专门针对研究南极物种蛋白质生命周期的显微镜和荧光工具的开发。这个博士项目将专注于一个显微镜平台的开发(第一年)以及荧光报告方法(第二年)。的确，低温下的显微技术带来了技术挑战。这些措施包括适当剂量的激光以维持细胞内的生理条件，以及严格控制显微镜工作台的温度，以便在一系列条件下观察样品。我们也期待着与显微镜材料本身的物理相关的挑战(不同部分的收缩，折射率的变化……)。一旦显微镜本身适应了极地条件，我们还必须开发在0℃左右工作的荧光工具。这些工具将用于研究影响蛋白质折叠和稳定性的因素，如粘度和细胞化学。将尝试两种测量细胞内粘度的方法，即单粒子跟踪(观察一种元素随时间的移动)和分子转子(根据细胞条件或多或少地旋转速度)。我们将结合显微镜和荧光技术在寒冷中将细胞表型与细胞所经历的温度变化联系起来(第3年)。这个项目是高度跨学科的，存在于生物学、物理学和工程学的交叉点，汇集了各种各样的科学技术。就EPSRC的战略和研究领域而言，该项目涉及以下内容：通过开发新的和独特的显微镜平台进行分析科学，通过研究基本蛋白质折叠动力学来研究生物物理学和软物质物理，或通过开发蛋白质跟踪和温度和粘度测量来开发传感器和仪器