Modulation of glutathione S-transferase and superoxide dismutase in response to heavy metals in brackish water flea Diaphanosoma celebensis

Abstract

Superoxide dismutase (SOD) and glutathione S-transferase (GST) serve as biomarkers of oxidative stress in aquatic organisms. However, the modulatory effects of heavy metals on the expression of small crustaceans are yet unknown. The acute toxicity of inorganic arsenic (As), cadmium (Cd), and lead (Pb) to the brackish water flea Diaphanosoma celebensis (4 days old) was evaluated to determine their sublethal concentrations. Modulations in the activities and transcription of SODs (Cu/Zn-SOD and Mn-SOD) and GSTs (mu, kappa, theta, and sigma) in response to sublethal concentrations of heavy metals were investigated for 48 h. The 48-h median lethal concentration (LC50) was determined to be 2.62 mg/L, 2.60 mg/L, and 0.37 mg/L for Cd, As, and Pb, respectively, and the order of toxicity in this species was Pb > Cd = As. SOD and GST activities were induced after exposure to all metals; in particular, GST activity was sensitively enhanced upon As exposure. Real-time reverse-transcriptase polymerase chain reaction showed that GST mu and GST sigma were specific for As, and GST kappa and GST theta for Cd. Mn-SOD mRNA expression was more sensitive to As. These findings suggest that D. celebensis SOD and GST may be involved in the antioxidant defense mechanisms to heavy metals. These genes may serve as useful biomarkers for risk assessment of heavy metal pollution in aquatic ecosystems. This study may be helpful for the better understanding of the molecular mode of action of heavy metals in small crustaceans and provide preliminary data for future studies evaluating the combined effects of heavy metal mixture.