Original scientific paper
https://doi.org/10.2478/10004-1254-61-2010-200
Presence of Ecotoxicologically Relevant Pgp and MRP Transcripts and Proteins in Cyprinid Fish
Roberta Sauerborn Klobučar
; Division for Marine and Environmental Research, Ruđer Bošković Institute, Zagreb, Croatia
Roko Žaja
; Division for Marine and Environmental Research, Ruđer Bošković Institute, Zagreb, Croatia
Damjan Franjević
; Division of Biology, Faculty of Science, University of Zagreb, Zagreb, Croatia
Anamarija Brozović
; Division for Molecular Biology, Ruđer Bošković Institute, Zagreb, Croatia
Tvrtko Smital
; Division for Marine and Environmental Research, Ruđer Bošković Institute, Zagreb, Croatia
Abstract
One of the most intriguing defence strategies which aquatic organisms developed through evolution is multixenobiotic resistance (MXR). The key mediators of MXR activity are ATP-binding cassette (ABC) transport proteins. They provide resistance of aquatic organisms by binding xenobiotics and extruding them from cells in an energy-dependent manner. Since Cyprinid fi sh species are common target in freshwater biomonitoring programs, we have studied the presence of two main MDR/MXR effl ux transporters Pglycoprotein
(Pgp, Abcb1) and MRP-like protein(s) (Abcc) in the liver of fi ve Cyprinid species: common carp, European chub, sneep, barbel, and silver prussian carp. Their presence was evaluated on the mRNA
and protein level. Various pairs of primers were designed to clone homologous fragments of MXR-related genes. At the protein level, we used Western blotting with specifi c monoclonal antibodies against human Pgp (Abcb1, Ab C219), MRP1 (Abcc1; Ab MRPm6) or MRP2 (Abcc2; Ab M2I-4). Transcripts of both key types of MXR transporters were identifi ed in all species examined and here we provide the phylogenetic analysis of new partial sequences. Immunochemical determinations with mammalian antibodies failed to identify the presence of MRP(s), but Pgp expression was found in all fi ve Cyprinid species. These results support that MXR is a defence system mediated by both Pgp and MRP types of ABC transport proteins.
Keywords
ABC proteins; aquatic organisms; multixenobiotic resistance; MXR; phylogenetic analysis; Western blot analysis
Hrčak ID:
53961
URI
Publication date:
16.6.2010.
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