Abstract
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.
Highlights
A critical need for instrumentation to study the effect of dissolved gases
Ischemia-reperfusion is a stress to tissues that occurs under a range of pathophysiologic conditions, and it is recognized that damage from hypoxia can occur both from the period of decreased energy production and at the time when O2 is replenished
After transformation of the inflow O2 using eq 4 and the transfer function generated with data obtained in the presence of potassium cyanide (KCN) to account for the delay and dispersion of the perifusion chamber, O2 consumption rate (OCR) was calculated from eq 5
Summary
A critical need for instrumentation to study the effect of dissolved gases. Oxygen (O2) is a fundamental determinant of cell survival and function in mammalian tissues. In most cells the majority of ATP is generated by oxidative phosphorylation, driven by a series of redox reactions in which O2 is the ultimate electron acceptor. Hypoxia is linked to many diseases including stroke, cancer and diabetic complications. In addition to O2, trace gases produced by cells (H2S, NO and CO) act as signals to regulate cellular and mitochondrial function. Ischemia-reperfusion injury is a condition common to many disease states and it is thought that during reoxygenation a burst of reactive O2 species (ROS)
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