Expression and molecular characterization of stress-responsive genes (hsp70 and Mn-sod) and evaluation of antioxidant enzymes (CAT and GPx) in heavy metal exposed freshwater ciliate, Tetmemena sp.
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Response of heavy metals namely cadmium (Cd) and copper (Cu) on the expression of stress responsive genes in the fresh water ciliate, Tetmemena sp. (single cell eukaryote) was studied. Stress responsive genes include heat shock protein genes and genes involved in antioxidant defence system. Quantitative real time PCR (qRT-PCR) was employed to evaluate the effects of Cd and Cu on the expression of cytosolic hsp70 and Mn-sod genes. Increase in the expression of these genes was observed after exposure with the heavy metals. The macronuclear cytosolic hsp70 and Mn-sod (SOD2) genes were also sequenced and characterized using various bioinformatics tools. In antioxidant defence system, the superoxide dismutase (SOD) family is a first line antioxidant enzyme group involved in catalysing reactive oxygen species (ROS) to hydrogen peroxide and molecular oxygen. Influence of Cd and Cu on the activity of SOD has already been reported by our group. Therefore, the enzymatic activities of antioxidant enzymes, catalase (CAT) and glutathione peroxidase (GPx) were studied in the presence of Cd and Cu and there was significant increase in activity of these enzymes in concentration dependent manner. This study suggests that cytosolic hsp70, Mn-sod and the antioxidant enzymes such as CAT and GPx can be used as effective molecular biomarkers for heavy metal toxicity and Tetmemena sp. can be used as potential model for understanding the molecular response to heavy metal contamination in aquatic ecosystems.
KeywordsAntioxidant enzymes Biomarkers Freshwater ciliate Heavy metal stress Stress genes Tetmemena sp.
The authors appreciate the facilities provided by the Principal, Acharya Narendra Dev College, University of Delhi for carrying out the present study. The support extended by the Principal, Maitreyi College, University of Delhi is thankfully acknowledged. We also thank Department of Zoology, University of Delhi, National Institute of Immunology and South Campus, University of Delhi for providing instrumentational facilities to perform the experiment. The work was also supported by the Senior Research Fellowships to SS from UGC (University Grants Commission) and Junior Research Fellowships to JSA and SM from CSIR (Council of Scientific and Industrial Research).
Compliance with ethical standards
Conflict of interest
The authors declare that there are no conflict of interest.
This article does not contain any studies conducted on humans and or vertebrate animals.
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