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Effects of diclazuril on the expression of enolase in second-generation merozoites of Eimeria tenella

Eimeria tenella is an obligate intracellular parasite of the chicken cecum; it brings huge economic loss to the chicken industry. Enolase is a multifunctional glycolytic enzyme involved in many processes of parasites, such as infection and migration. In this study, the effect of diclazuril on the ex...

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Autores principales: Zhou, Bian-hua, Ding, Hai-yan, Yang, Jing-yun, Chai, Jun, Guo, Hong-wei, Wang, Hong-wei
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7705050/
https://www.ncbi.nlm.nih.gov/pubmed/33248555
http://dx.doi.org/10.1016/j.psj.2020.09.068
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author Zhou, Bian-hua
Ding, Hai-yan
Yang, Jing-yun
Chai, Jun
Guo, Hong-wei
Wang, Hong-wei
author_facet Zhou, Bian-hua
Ding, Hai-yan
Yang, Jing-yun
Chai, Jun
Guo, Hong-wei
Wang, Hong-wei
author_sort Zhou, Bian-hua
collection PubMed
description Eimeria tenella is an obligate intracellular parasite of the chicken cecum; it brings huge economic loss to the chicken industry. Enolase is a multifunctional glycolytic enzyme involved in many processes of parasites, such as infection and migration. In this study, the effect of diclazuril on the expression of enolase in second-generation merozoites of E. tenella (EtENO) was reported. The prokaryotic expression plasmid pET-28a-EtENO was constructed and transformed into Escherichia coli BL21 (DE3). Then, it was subjected to expression under the induction of isopropyl-β-D-1-thiogalactopyranoside. The expressed products were identified and purified. The purified EtENO protein was used for antibody preparation. The EtENO mRNA and protein expression levels were analyzed via real-time PCR and Western blotting. Localization of EtENO on the merozoites was examined by immunofluorescence technique. The mRNA and protein expression levels of EtENO were decreased by 36.3 and 40.36%, respectively, by diclazuril treatment. EtENO distributed in the surface, cytoplasm, and nucleus of the infected/control group. With diclazuril treatment, it was significantly reduced in the surface and cytoplasm and even disappeared in the nucleus of the infected/diclazuril group. These observations suggested that EtENO may play an important role in mechanism of diclazuril anticoccidial action and be a potential drug target for the intervention with E. tenella infection.
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spelling pubmed-77050502020-12-08 Effects of diclazuril on the expression of enolase in second-generation merozoites of Eimeria tenella Zhou, Bian-hua Ding, Hai-yan Yang, Jing-yun Chai, Jun Guo, Hong-wei Wang, Hong-wei Poult Sci Genetics and Molecular Biology Eimeria tenella is an obligate intracellular parasite of the chicken cecum; it brings huge economic loss to the chicken industry. Enolase is a multifunctional glycolytic enzyme involved in many processes of parasites, such as infection and migration. In this study, the effect of diclazuril on the expression of enolase in second-generation merozoites of E. tenella (EtENO) was reported. The prokaryotic expression plasmid pET-28a-EtENO was constructed and transformed into Escherichia coli BL21 (DE3). Then, it was subjected to expression under the induction of isopropyl-β-D-1-thiogalactopyranoside. The expressed products were identified and purified. The purified EtENO protein was used for antibody preparation. The EtENO mRNA and protein expression levels were analyzed via real-time PCR and Western blotting. Localization of EtENO on the merozoites was examined by immunofluorescence technique. The mRNA and protein expression levels of EtENO were decreased by 36.3 and 40.36%, respectively, by diclazuril treatment. EtENO distributed in the surface, cytoplasm, and nucleus of the infected/control group. With diclazuril treatment, it was significantly reduced in the surface and cytoplasm and even disappeared in the nucleus of the infected/diclazuril group. These observations suggested that EtENO may play an important role in mechanism of diclazuril anticoccidial action and be a potential drug target for the intervention with E. tenella infection. Elsevier 2020-10-07 /pmc/articles/PMC7705050/ /pubmed/33248555 http://dx.doi.org/10.1016/j.psj.2020.09.068 Text en © 2020 Published by Elsevier Inc. on behalf of Poultry Science Association Inc. http://creativecommons.org/licenses/by-nc-nd/4.0/ This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Genetics and Molecular Biology
Zhou, Bian-hua
Ding, Hai-yan
Yang, Jing-yun
Chai, Jun
Guo, Hong-wei
Wang, Hong-wei
Effects of diclazuril on the expression of enolase in second-generation merozoites of Eimeria tenella
title Effects of diclazuril on the expression of enolase in second-generation merozoites of Eimeria tenella
title_full Effects of diclazuril on the expression of enolase in second-generation merozoites of Eimeria tenella
title_fullStr Effects of diclazuril on the expression of enolase in second-generation merozoites of Eimeria tenella
title_full_unstemmed Effects of diclazuril on the expression of enolase in second-generation merozoites of Eimeria tenella
title_short Effects of diclazuril on the expression of enolase in second-generation merozoites of Eimeria tenella
title_sort effects of diclazuril on the expression of enolase in second-generation merozoites of eimeria tenella
topic Genetics and Molecular Biology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7705050/
https://www.ncbi.nlm.nih.gov/pubmed/33248555
http://dx.doi.org/10.1016/j.psj.2020.09.068
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