Transcriptional responses of alpha‐ and rho‐class glutathione S‐transferase genes in the liver of three freshwater fishes intraperitoneally injected with microcystin‐LR: Relationship of inducible expression and tolerance

Abstract

Rho-class glutathione S-transferase (GST) is found only in teleost fish with no homologues in mammals. Silver carp (Hypophthalmichthys molitrix), grass carp (Ctenopharyngodon idellus), and Nile tilapia (Oreochromis nilotica) are three warm freshwater fishes with differential tolerance to microcystin-LR (MC-LR): Nile tilapia has a little higher tolerance than silver carp, but both have much higher tolerance than grass carp. Full-length cDNAs encoding the rho-class GST were cloned and sequenced from the livers of the three fishes. The silver carp, grass carp, and Nile tilapia rho-class GST cDNAs were 1078, 1104, and 904 bp in length, respectively, and all contained an open-reading frame (ORF) of 681 bp (encoding a polypeptide of 226 amino acids). Using beta-actin as an external control, semiquantitative RT-PCR was conducted to determine the constitutive and inducible expression level of alpha- and rho-class GST genes among the three fishes (6-12 g) intraperitoneally injected with MC-LR (50 mug kg(-1) body weight). Liver mRNA expression levels of alpha-class GST gene were found to be higher than those of rho-class GST gene in both exposed and control fish of silver carp and grass carp, whereas liver mRNA expression levels of rho-class GST gene were higher than those of alpha -class GST gene in both exposed and control fish of Nile tilapia. Increased liver transcription of GST isoforms was detected at 8-h postinjection of MC-LR in silver carp (alpha- and rho-class GST) and Nile tilapia (rho-class GST), and at 24-h postinjection in silver carp (alpha-class GST) and Nile tilapia (alpha-class GST), but an increase in liver transcription neither of alpha-class GST nor of rho-class GST was detected in grass carp at either 8-h or 24-h postinjection. The inducible expression of the liver GST gene showed a close relationship with their tolerance to MC-LR: high-resistant fish (phytoplanktivorous silver carp and Nile tilapia) had inducible liver expression of either alpha- or rho-class GST, and the high-sensitive fish (herbivorous grass carp) had no inducible liver expression of either one. We suggest that inducible expression (instead of constitutive expression) of the liver GST gene should play an important role in the tolerance to microcystin exposure, and that in addition to alpha-class GST, high-liver expression of rho-class GST gene might have facilitated the evolutionary radiation of teleost fish.