Abstract

The glutathione-S-transferase (GST) superfamily includes seven classes, and different classes have different functions. GST superfamily members function in various processes including detoxification of xenobiotics, protection against oxidative damage, and intracellular transport of hormones, endogenous metabolites, and exogenous chemicals. Herein, to elucidate the tissue-specific expression pattern of GSTs in response to hypoxia stress, which induces cell death, we investigated the expression of GSTs in response to hypoxia and reoxygenation in oriental river prawn, Macrobrachium nipponense. Full-length cDNAs of two δ class GSTs were cloned from the hepatopancreas, and named MnGST-1 and MnGST-2 based on the established GST nomenclature system. Expression profiles of both GSTs in various tissues were different under acute and chronic experimental hypoxia stress conditions, suggesting that both respond strongly to hypoxia-induced oxidative stress. However, the intensity of responses to hypoxia and reoxygenation were different in different tissues. During acute hypoxia stress, MnGST-1 responds earlier than MnGST-2 in the hepatopancreas and gill, but more slowly in muscle. By contrast, during chronic hypoxia stress, MnGST-2 plays a more important role in the hepatopancreas and gill than MnGST-1.

Highlights

  • The level of dissolved oxygen (DO) is a major indicator of water quality, in the prawn farming industry

  • Under hypoxia conditions, accumulated electrons contribute to the formation of damaging reactive oxygen species (ROS) [5], and reoxygenation of hypoxic tissues can lead to ROS generation [6]

  • To protect themselves against damage by ROS, aerobic organisms have evolved a set of antioxidant defense systems, including antioxidant enzymes, such as superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), and glutathione S-transferase (GST) [8,9,10,11]

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Summary

Introduction

The level of dissolved oxygen (DO) is a major indicator of water quality, in the prawn farming industry. Under hypoxia conditions, accumulated electrons contribute to the formation of damaging reactive oxygen species (ROS) [5], and reoxygenation of hypoxic tissues can lead to ROS generation [6]. ROS damage important biomolecules including DNA, proteins and lipids. M. nipponense is an important commercial prawn species that is widely distributed in freshwater and low-salinity estuarine regions in China and other Asian countries. Understanding the molecular mechanisms underpinning the stress responses to hypoxia and reoxygenation in M. nipponense is essential for the sustained development of prawn aquaculture. We cloned and characterized full-length MnGST-1 and MnGST-2 cDNAs from M. nipponense, and quantified their tissue-specific mRNA expression levels prawns in response to hypoxia and reoxygenation. We assayed the enzyme activity of the MnGST-1 and MnGST-2 proteins in prawns under hypoxia and reoxygenation conditions

Prawn MnGST-1 and MnGST-2 Coding Sequences
Tissue Distribution of MnGST-1 and MnGST-2
Experimental Animals and Hypoxic Treatment
Cloning of MnGST-1 and MnGST-2 cDNAs
Nucleotide Sequence and Bioinformatics Analyses
Semiquantitative Analysis of MnGST-1 and MnGST-2 Expression
Enzyme Activity Assay
Statistical Analysis
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