Fluidics system for resolving concentration-dependent effects of dissolved gases on tissue metabolism
Varun Kamat,
Brian M Robbings,
Seung-Ryoung Jung,
John Kelly,
James B Hurley,
Kenneth P Bube,
Ian R Sweet
Affiliations
Varun Kamat
University of Washington Medicine Diabetes Institute, University of Washington, Seattle, United States
Brian M Robbings
University of Washington Medicine Diabetes Institute, University of Washington, Seattle, United States; Department of Biochemistry, University of Washington, Seattle, United States
Seung-Ryoung Jung
University of Washington Medicine Diabetes Institute, University of Washington, Seattle, United States
Oxygen (O2) and other dissolved gases such as the gasotransmitters H2S, CO, and NO affect cell metabolism and function. To evaluate effects of dissolved gases on processes in tissue, we developed a fluidics system that controls dissolved gases while simultaneously measuring parameters of electron transport, metabolism, and secretory function. We use pancreatic islets, retina, and liver from rodents to highlight its ability to assess effects of O2 and H2S. Protocols aimed at emulating hypoxia–reperfusion conditions resolved a previously unrecognized transient spike in O2 consumption rate (OCR) following replenishment of O2, and tissue-specific recovery of OCR following hypoxia. The system revealed both inhibitory and stimulatory effects of H2S on insulin secretion rate from isolated islets. The unique ability of this new system to quantify metabolic state and cell function in response to precise changes in dissolved gases provides a powerful platform for cell physiologists to study a wide range of disease states.
