Aquatic organisms are frequently exposed to multiple stressors including low dissolved oxygen (O) and metals such as cadmium (Cd). Reduced O concentration and Cd exposure alter cellular function in part by impairing energy metabolism and dysregulating reactive oxygen species (ROS) homeostasis. However, little is known about the role of mitochondrial glycerol 3-phosphate dehydrogenase (mGPDH) in ROS homeostasis in fish and its response to environmental stress. In this study, mGPDH activity and the effects of anoxia-reoxygenation (A-RO) and Cd on ROS (as hydrogen peroxide, HO) emission in rainbow trout liver mitochondria during oxidation of glycerol 3-phosphate (G3P) were probed. Trout liver mitochondria exhibited low mGPDH activity that supported a low respiratory rate but substantial HO emission rate. Cd evoked a low concentration stimulatory-high concentration inhibitory HO emission pattern that was blunted by A-RO. At specific redox centers, Cd suppressed HO emission from site I, but stimulated emission from sites III and G. In contrast, A-RO stimulated HO emission from site I following 15 min exposure and augmented Cd-stimulated emission from site II after 30 min exposure but did not alter the rate of HO emission from sites III and G. Additionally, Cd neither altered the activities of catalase, glutathione peroxidase, or thioredoxin reductase nor the concentrations of total glutathione, reduced glutathione, or oxidized glutathione. Overall, this study indicates that oxidation of G3P drives ROS production from mGPDH and complexes I, II and III, whereas Cd directly modulates redox sites but not antioxidant defense systems to alter mitochondrial HO emission.
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