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Gene expression analysis of ABC transporters in a resistant Cooperia oncophora isolate following in vivo and in vitro exposure to macrocyclic lactones

Published online by Cambridge University Press:  02 January 2013

J. DE GRAEF ,
J. DEMELER ,
P. SKUCE ,
M. MITREVA ,
G. VON SAMSON-HIMMELSTJERNA ,
J. VERCRUYSSE ,
E. CLAEREBOUT  and
P. GELDHOF
J. DE GRAEF
Affiliation:
Laboratory for Parasitology, Faculty of Veterinary Medicine, Ghent University, Salisburylaan 133, 9820 Merelbeke, Belgium
J. DEMELER
Affiliation:
Institute for Parasitology and Tropical Veterinary Medicine, Freie Universität Berlin, Königsweg 67, 14163 Berlin, Germany
P. SKUCE
Affiliation:
Moredun Research Institute, Pentlands Science Park, Bush Loan, Penicuik, Midlothian, Scotland, UK
M. MITREVA
Affiliation:
The Genome Institute, Washington University School of Medicine, 4444 Forest Park Boulevard, St Louis, MO 63108, USA Department of Genetics, Washington University School of Medicine, 4444 Forest Park Boulevard, St Louis, MO 63108, USA
G. VON SAMSON-HIMMELSTJERNA
Affiliation:
Institute for Parasitology and Tropical Veterinary Medicine, Freie Universität Berlin, Königsweg 67, 14163 Berlin, Germany
J. VERCRUYSSE
Affiliation:
Laboratory for Parasitology, Faculty of Veterinary Medicine, Ghent University, Salisburylaan 133, 9820 Merelbeke, Belgium
E. CLAEREBOUT
Affiliation:
Laboratory for Parasitology, Faculty of Veterinary Medicine, Ghent University, Salisburylaan 133, 9820 Merelbeke, Belgium
P. GELDHOF*
Affiliation:
Laboratory for Parasitology, Faculty of Veterinary Medicine, Ghent University, Salisburylaan 133, 9820 Merelbeke, Belgium
*
*Corresponding author: Laboratory for Parasitology, Faculty of Veterinary Medicine, Ghent University, Salisburylaan 133, 9820 Merelbeke, Belgium. Tel: +32 (09) 2647517. Fax: +32 (09) 2647496. E-mail: peter.geldhof@ugent.be
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Summary

Members of the ATP-binding cassette (ABC) transporter family (P-glycoproteins, Half-transporters and Multidrug Resistant Proteins) potentially play a role in the development of anthelmintic resistance. The aim of this study was to investigate the possible involvement of ABC transporters in anthelmintic resistance in the bovine parasite, Cooperia oncophora. Partial sequences of 15 members of the ABC transporter protein family were identified, by mining a transcriptome dataset combined with a degenerate PCR approach. Reverse transcriptase PCR showed that most of the ABC transporters identified were constitutively transcribed throughout the life cycle of C. oncophora. Constitutive differences in gene transcript levels between a susceptible and resistant isolate were only observed for Con-haf-9 and Con-mrp-1 in eggs of the resistant isolate, while no differences were observed in L3 or the adult life stage. Analysis of resistant adult worms, collected from calves 14 days after treatment with either ivermectin or moxidectin, showed a significant 3- to 5-fold increase in the transcript levels of Con-pgp-11 compared to non-exposed worms. Interestingly, a 4-fold transcriptional up-regulation of Con-pgp-11 was also observed in L3 of the resistant isolate, after in vitro exposure to different concentrations of ivermectin, whereas this effect was not observed in exposed L3 of the susceptible isolate. The results suggest that the worms of this particular resistant isolate have acquired the ability to up-regulate Con-pgp-11 upon exposure to macrocyclic lactones. Further work is needed to understand the genetic basis underpinning this process and the functional role of PGP-11.

Keywords

Cooperia oncophoraivermectin resistanceABC transportersPGPHAFMRP
Type
Research Article
Information
Parasitology , Volume 140 , Issue 4 , April 2013 , pp. 499 - 508
Copyright
Copyright © Cambridge University Press 2013

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