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

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

The primary outputs from the CDT will be cohorts of highly qualified, interdisciplinary postgraduates who are experts in a wide range of sensing activities. They will benefit from a world leading training experience that recognises sensor research as an academic discipline in its own right. The students will be taught in all aspects of Sensor Technologies, ranging from the physical and chemical principles of sensing, to sensor design, data capture and processing, all the way to applications and opportunities for commercialisation, with a strong focus in entrepreneurship, technology translation and responsible leadership. Students will learn in extensive team and cohort engaging activities, and have access to cutting-edge expertise and infrastructure. 90 academics from 15 different departments participate in the programme and more than 40 industrial partners are actively involved in delivering research and business leadership training, offering perspectives for impact and translation and opportunities for internships and secondments. End users associated with the CDT will benefit from the availability of outstanding, highly qualified and motivated PhD students, access to shared infrastructure, and a huge range of academic and industrial contacts.

Immediate beneficiaries of our CDT will be our core industrial consortium partners (MedImmune, Alphasense, Fluidic Analytics, ioLight, NokiaBell, Cambridge Display Technologies, Teraview, Zimmer and Peacock, Panaxium, Silicon Microgravity, etc., see various LoS) who incorporate our cross-leverage funding model into their corporate research strategies. Small companies and start-ups particularly benefit from the flexibility of the partnerships we can offer. We will engage through weekly industry seminars and monthly Sensor Cafés, where SME employees can interact directly with the CDT students and PIs, provide training in topical areas, and, in turn, gain themselves access to CDT infrastructure and training. Ideas can be rapidly tested through industrially focused miniprojects and promising leads developed into funded PhD programmes, for which leveraged funding is available through the CDT.

Government departments and large research initiatives are formally connected to the CDT, including the Department for the Environment, Food and Rural Affairs (DEFRA); the Cambridge Centre for Smart Infrastructure and Construction (CSIC); the Centre for Global Equality (CGE); the National Physics Laboratory (NPL); the British Antarctic Survey (BAS), who all push our CDT to generate impacts that are in the public interest and relevant for a healthy and sustainable future society. With their input, we will tackle projects on assisted living technologies for the ageing population, diagnostics of environmental toxins in the developing world, and sensor technologies that help replace the use of animals in research. Developing countries will benefit through our emphasis on open technologies / open innovation and our exploration of responsible, ethical, and transparent business models. In the UK, our CDT will engage directly with the public sector and national policy makers and regulators (DEFRA, and the National Health Service - NHS) and, with their input, students are trained on impact and technology translation, ethics, and regulatory frameworks.