USAP-DC

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 Merit While 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 Impacts The 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.

Person	Role
Buckley, Bradley	Investigator and contact
Kueltz, Dietmar	Co-Investigator

Antarctic Organisms and Ecosystems

Award # 2322117

USAP-2322117_1

Buckley_Data_Management_Plan.pdf