Mitochondrial membrane potential is a suitable candidate for assessing pollution toxicity in fish

Abstract

Fish inhabiting polluted estuaries are highly exposed to severe stress characterized by an oxidant–antioxidant imbalance. The aim of the study was to explore the use of stress parameters such as adenosine triphosphate/adenosine diphosphate (ATP/ADP) ratio, mitochondrial membrane potential (∆ψm) and total protein expression patterns as biomarkers against oxidant exposures in hepatocytes of Mugil cephalus living in either a contaminated (Test; Ennore) or uncontaminated (Control; Kovalam) estuary. Earlier, the pollutant stress impact was determined through light and electron microscopy studies. The ATP/ADP ratio was measured using high performance liquid chromatography; ∆ψm by fluorescent probe 5, 5′, 6, 6′-tetrachloro-1, 1′, 3, 3′-tetraethyl benzimidazolcarbocyanine iodide (JC-1) dye and total protein expression patterns by protein profiling. The preponderance of stress impact was confirmed through microscopy studies that featured cytological alterations, disturbances in the surface morphology and in the cell organelles at the ultrastructural levels. Hepatocytes of test fish demonstrated a decrease in ATP and an increase in ADP and thereby alteration in ATP/ADP ratio ( p < 0.05; 20.75%). A significant disturbance ( p < 0.05; 26.57%) in ∆ψm with a ratio of J-aggregates/JC-1 monomer of 1 was observed for test fish hepatocytes compared to control group with a J-aggregates/JC-1 monomer ratio of 1.5. Quantitative assessment of protein expression levels also revealed enhanced induction of both low and high molecular weight proteins in test fish hepatocytes. The findings highlight the use of these parameters as the highly sensitive biomarkers in response to contaminant exposure compared to the routinely used antioxidant and oxidant stress parameters in biomonitoring programs. Among the measured parameters, the determination of ∆ψm may be suggested as a novel candidate as a biomarker because of its greater specificity and rapid quantitative risk assessment of pollutant exposures.