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Research article - Peer-reviewed, 2016

Differential expression of beta-tubulin isotypes in different life stages of Parascaris spp after exposure to thiabendazole

Tyden, Eva; Skarin, Moa; Andersson Franko, Mikael; Sjöblom, Matilda; Höglund, Johan

Abstract

Anthelmintic resistance (AR) to macrocyclic lactones (ML) has been described in Parascaris of horses world-wide. In contrast, benzimidazoles (BZ) are still effective, although reduced efficacy to this drug class was recently reported. The mode of action of BZ is binding to beta-tubulin, which prevents polymerisation of microtubules. In this study, beta-tubulin gene expression of isotypes 1 and 2 was investigated at seven time points (0, 6, 24, 72, 96 and 120h) during embryogenesis and in adult worms. In addition, an in ovo larval developmental test was developed to study beta-tubulin gene expression of both isotypes in parasacaris eggs after exposure to different concentrations of thiabendazole (TBZ) for five days at 25 degrees C. A strong pattern of differential expression of beta-tubulin and isotype 1 was observed in all stages, while isotype 2 expression was mainly found at an early phase of the embryogenesis. For isotype 1, a 5-fold increase was observed during the first 48 h, but gene expression gradually decreased after 72, 96 and 120 h. Isotype 2 was only expressed during the first 24 h, followed by a 130-fold decrease at (time points) 72, 96 and 120 h. The in ovo larval developmental test, in which we exposed initially unembryonated eggs to increased concentrations of TBZ, did affect isotype I gene expression but not isotype 2. This assumes that each isotype has specific functions in different life stages. This is in agreement with the 'multi-tubulin' hypothesis, which states that different tubulin isotypes are required for specialised microtubule functions. Isotype I is the most likely drug target for BZs, as isotype 2 was only expressed at very low levels later in development. Increasing concentrations of TBZ altered beta-tubulin isotype 1 gene expression after exposure of the eggs for five days, but this was not seen for isotype 2. (C) 2016 The Authors. Published by Elsevier B.V. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

Keywords

Parascaris spp; beta-tubulin; Gene expression; Life stages; Embryogenesis; Thiabendazole

Published in

Molecular and Biochemical Parasitology
2016, volume: 205, number: 1-2, pages: 22-28

Authors' information

Swedish University of Agricultural Sciences, Department of Biomedical Science and Veterinary Public Health
Swedish University of Agricultural Sciences, Department of Biomedical Science and Veterinary Public Health
Andersson Franko, Mikael
Swedish University of Agricultural Sciences, Department of Economics
Sjöblom, Matilda
Swedish University of Agricultural Sciences, Department of Biomedical Science and Veterinary Public Health

Associated SLU-program

Future Animal Health and Welfare (until Jan 2017)

UKÄ Subject classification

Other Basic Medicine

Publication Identifiers

DOI: https://doi.org/10.1016/j.molbiopara.2016.02.004

URI (permanent link to this page)

https://res.slu.se/id/publ/73781