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Characterization of an MLP Homologue from Haemaphysalis longicornis (Acari: Ixodidae) Ticks

Members of the cysteine-rich protein (CRP) family are known to participate in muscle development in vertebrates. Muscle LIM protein (MLP) belongs to the CRP family and has an important function in the differentiation and proliferation of muscle cells. In this study, the full-length cDNA encoding MLP...

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Autores principales: Luo, Jin, Shen, Hui, Ren, Qiaoyun, Guan, Guiquan, Zhao, Bo, Yin, Hong, Chen, Ronggui, Zhao, Hongying, Luo, Jianxun, Li, Xiangrui, Liu, Guangyuan
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7238268/
https://www.ncbi.nlm.nih.gov/pubmed/32295244
http://dx.doi.org/10.3390/pathogens9040284
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author Luo, Jin
Shen, Hui
Ren, Qiaoyun
Guan, Guiquan
Zhao, Bo
Yin, Hong
Chen, Ronggui
Zhao, Hongying
Luo, Jianxun
Li, Xiangrui
Liu, Guangyuan
author_facet Luo, Jin
Shen, Hui
Ren, Qiaoyun
Guan, Guiquan
Zhao, Bo
Yin, Hong
Chen, Ronggui
Zhao, Hongying
Luo, Jianxun
Li, Xiangrui
Liu, Guangyuan
author_sort Luo, Jin
collection PubMed
description Members of the cysteine-rich protein (CRP) family are known to participate in muscle development in vertebrates. Muscle LIM protein (MLP) belongs to the CRP family and has an important function in the differentiation and proliferation of muscle cells. In this study, the full-length cDNA encoding MLP from Haemaphysalis longicornis (H. longicornis; HLMLP) ticks was obtained by 5′ rapid amplification of cDNA ends (RACE). To verify the transcriptional status of MLP in ticks, HLMLP gene expression was assessed during various developmental stages by real-time PCR (RT-PCR). Interestingly, HLMLP expression in the integument was significantly (P < 0.01) higher than that observed in other tested tissues of engorged adult ticks. In addition, HLMLP mRNA levels were significantly downregulated in response to thermal stress at 4 °C for 48 h. Furthermore, recombinant HLMLP was expressed in Escherichia coli, and Western blot analysis showed that rabbit antiserum against H. longicornis adults recognized HLMLP and MLPs from different ticks. Ten 3-month-old rabbits that had never been exposed to ticks were used for the immunization and challenge experiments. The rabbits were divided into two groups of five rabbits each, where rabbits in the first group were immunized with HLMLP, while those in the second group were immunized with phosphate-buffered saline (PBS) diluent as controls. The vaccination of rabbits with the recombinant HLMLP conferred partial protective immunity against ticks, resulting in 20.00% mortality and a 17.44% reduction in the engorgement weight of adult ticks. These results suggest that HLMLP is not ideal as a candidate for use in anti-tick vaccines. However, the results of this study generated novel information on the MLP gene in H. longicornis and provide a basis for further investigation of the function of this gene that could potentially lead to a better understanding of the mechanism of myofiber determination and transformation
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spelling pubmed-72382682020-06-02 Characterization of an MLP Homologue from Haemaphysalis longicornis (Acari: Ixodidae) Ticks Luo, Jin Shen, Hui Ren, Qiaoyun Guan, Guiquan Zhao, Bo Yin, Hong Chen, Ronggui Zhao, Hongying Luo, Jianxun Li, Xiangrui Liu, Guangyuan Pathogens Article Members of the cysteine-rich protein (CRP) family are known to participate in muscle development in vertebrates. Muscle LIM protein (MLP) belongs to the CRP family and has an important function in the differentiation and proliferation of muscle cells. In this study, the full-length cDNA encoding MLP from Haemaphysalis longicornis (H. longicornis; HLMLP) ticks was obtained by 5′ rapid amplification of cDNA ends (RACE). To verify the transcriptional status of MLP in ticks, HLMLP gene expression was assessed during various developmental stages by real-time PCR (RT-PCR). Interestingly, HLMLP expression in the integument was significantly (P < 0.01) higher than that observed in other tested tissues of engorged adult ticks. In addition, HLMLP mRNA levels were significantly downregulated in response to thermal stress at 4 °C for 48 h. Furthermore, recombinant HLMLP was expressed in Escherichia coli, and Western blot analysis showed that rabbit antiserum against H. longicornis adults recognized HLMLP and MLPs from different ticks. Ten 3-month-old rabbits that had never been exposed to ticks were used for the immunization and challenge experiments. The rabbits were divided into two groups of five rabbits each, where rabbits in the first group were immunized with HLMLP, while those in the second group were immunized with phosphate-buffered saline (PBS) diluent as controls. The vaccination of rabbits with the recombinant HLMLP conferred partial protective immunity against ticks, resulting in 20.00% mortality and a 17.44% reduction in the engorgement weight of adult ticks. These results suggest that HLMLP is not ideal as a candidate for use in anti-tick vaccines. However, the results of this study generated novel information on the MLP gene in H. longicornis and provide a basis for further investigation of the function of this gene that could potentially lead to a better understanding of the mechanism of myofiber determination and transformation MDPI 2020-04-14 /pmc/articles/PMC7238268/ /pubmed/32295244 http://dx.doi.org/10.3390/pathogens9040284 Text en © 2020 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Luo, Jin
Shen, Hui
Ren, Qiaoyun
Guan, Guiquan
Zhao, Bo
Yin, Hong
Chen, Ronggui
Zhao, Hongying
Luo, Jianxun
Li, Xiangrui
Liu, Guangyuan
Characterization of an MLP Homologue from Haemaphysalis longicornis (Acari: Ixodidae) Ticks
title Characterization of an MLP Homologue from Haemaphysalis longicornis (Acari: Ixodidae) Ticks
title_full Characterization of an MLP Homologue from Haemaphysalis longicornis (Acari: Ixodidae) Ticks
title_fullStr Characterization of an MLP Homologue from Haemaphysalis longicornis (Acari: Ixodidae) Ticks
title_full_unstemmed Characterization of an MLP Homologue from Haemaphysalis longicornis (Acari: Ixodidae) Ticks
title_short Characterization of an MLP Homologue from Haemaphysalis longicornis (Acari: Ixodidae) Ticks
title_sort characterization of an mlp homologue from haemaphysalis longicornis (acari: ixodidae) ticks
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7238268/
https://www.ncbi.nlm.nih.gov/pubmed/32295244
http://dx.doi.org/10.3390/pathogens9040284
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