We are requesting funds to purchase a Seahorse XF96 Extracellular Flux Analyzer, an instrument that profiles the metabolic activity and bioenergetic state of cells non-invasively, in minutes, and in a medium/high- throughput manner. The Seahorse XF96 Extracellular Flux Analyzer uses a simple microplate format to simultaneously measure oxygen consumption (cellular respiration) and extracellular acidification rates, in real time. Combined with the use of different substrates for energy and various inhibitors of specific enzymes and metabolic pathways, which can be delivered automatically via four integrated injection ports, the Seahorse Extracellular Flux Analyzer provides an unprecedented, thorough and fast assessment of oxidative phosphorylation, glycolysis, nutrient utilization, fatty acid oxidation, coupled vs. uncoupled respiration, and spare respiratory capacity of cells. There is currently no other single instrument that can perform the same range of metabolic assays in a 96-well format. Mitochondria function and proper substrate utilization are central to most physiologic processes important for human health, including growth, energy homeostasis, capacity for physical activity, and aging. Mitochondrial dysfunction has been implicated in the initiation, development and/or pathogenesis of a wide variety of diseases, including metabolic diseases (e.g. insulin resistance and development of type 2 diabetes), cancer, neurodegenerative diseases, and cardiovascular disease. The requested Seahorse XF96 Extracellular Flux Analyzer will enable a minimum of 12 investigators, which currently have no access to a similar instrument or technology, to gain insights into the cellular metabolism of a wide range of cells and organisms used to model physiological and pathological processes (including primary hepatocytes, adipocytes, neurons, cardiomyocytes, cancer cells, myotubes, and worms). The key elements of the Seahorse XF96 Extracellular Flux Analyzer that make it ideally suited for the specific needs of the users are the ability of the instrument to: 1) monitor attached cells (a crucial consideration for neurons, adipocytes, hepatocytes, and myotubes, used by 7 of 12 investigators);2) obtain reliable measurements from small numbers of cells (crucial for 6 users that derive primary cells from mouse genetic models;and, 3) offer a medium/high throughput format, thereby enabling drug screening (5 projects) and evaluation of responses to a wide range of physiologic signals, environmental insults, and genetic alterations (important for all projects). The Seahorse XF96 Extracellular Flux Analyzer is unsurpassed in the combination of features that make it ideally suited to the studies of the current users. The instrument will be placed in a multi-user appropriate environment, readily accessible to all users, and available to new users. In summary, by providing a comprehensive assessment of cellular metabolism and mitochondrial function, the Seahorse XF96 Extracellular Flux Analyzer will facilitate and enhance our research on metabolism and metabolic diseases, cancer, neurodegenerative diseases, aging, calorie restriction, and diseases of protein folding.
|Gantner, Marin L; Hazen, Bethany C; Conkright, Juliana et al. (2014) GADD45? regulates the thermogenic capacity of brown adipose tissue. Proc Natl Acad Sci U S A 111:11870-5|