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Toxins 2015, 7(6), 2096-2120; doi:10.3390/toxins7062096

Glutathione Transferases Responses Induced by Microcystin-LR in the Gills and Hepatopancreas of the Clam Venerupis philippinarum

1
CIIMAR/CIMAR—Interdisciplinary Centre of Marine and Environmental Research, University of Porto, Rua dos Bragas 289, Porto 4050-123, Portugal
2
IPATIMUP—Institute of Molecular Pathology and Immunology of the University of Porto, Porto 4200-465, Portugal
3
Instituto de Investigação e Inovação em Saúde, University of Porto, Porto 4200-135, Portugal
4
Faculty of Medicine, University of Porto, Porto 4200-319, Portugal
5
Department of Biology, Faculty of Sciences, Porto University, Rua do Campo Alegre, Porto 4069-007, Portugal
*
Author to whom correspondence should be addressed.
Academic Editor: Elizabeth D. Hilborn
Received: 6 May 2015 / Accepted: 30 May 2015 / Published: 9 June 2015
(This article belongs to the Special Issue Bioactivity and Toxicity in Marine Cyanobacteria)
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Abstract

A multi-method approach was employed to compare the responses of Glutatione Transferases (GSTs) in the gills and hepatopancreas of Venerupis philippinarum to microcystins (MCs) toxicity. In this way, using the cytosolic fraction, the enzymatic activity of GSTs, superoxide dismutase (SOD), serine/threonine protein phosphatases (PPP2) along with the gene expression levels of four GST isoforms (pi, mu, sigma1, sigma2) were investigated in both organs of the clams exposed for 24 h to 10, 50 and 100 μg L1 of MC-LR. Cytosolic GSTs (cGSTs) from both organs of the high dose exposed clams were purified by glutathione-agarose affinity chromatography, characterized kinetically and the changes in the expression of cGSTs of the gills identified using a proteomic approach. MC-LR caused an increase in GST enzyme activity, involved in conjugation reactions, in both gills and hepatopancreas (100 μg L1 exposure). SOD activity, an indicator of oxidative stress, showed significantly elevated levels in the hepatopancreas only (50 and 100 μg L1 exposure). No significant changes were found in PPP2 activity, the main target of MCs, for both organs. Transcription responses revealed an up-regulation of sigma2 in the hepatopancreas at the high dose, but no significant changes were detected in the gills. Kinetic analysis evidenced differences between gills of exposed and non-exposed extracts. Using proteomics, qualitative and quantitative differences were found between the basal and inducible cGSTs. Overall, results suggest a distinct role of GST system in counteracting MCs toxicity between the gills and the hepatopancreas of V. philippinarum, revealing different roles between GST isoforms within and among both organs. View Full-Text
Keywords: microcystins; glutathione transferases; enzyme activity; proteomics; real-time PCR; V. philippinarum; detoxification; biomarker microcystins; glutathione transferases; enzyme activity; proteomics; real-time PCR; V. philippinarum; detoxification; biomarker
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).

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MDPI and ACS Style

Carneiro, M.; Reis, B.; Azevedo, J.; Campos, A.; Osório, H.; Vasconcelos, V.; Martins, J.C. Glutathione Transferases Responses Induced by Microcystin-LR in the Gills and Hepatopancreas of the Clam Venerupis philippinarum. Toxins 2015, 7, 2096-2120.

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