Cargando…
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...
Autores principales: | , , , , , , , , , , |
---|---|
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 |
_version_ | 1783536506994950144 |
---|---|
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 |
format | Online Article Text |
id | pubmed-7238268 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
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 |
work_keys_str_mv | AT luojin characterizationofanmlphomologuefromhaemaphysalislongicornisacariixodidaeticks AT shenhui characterizationofanmlphomologuefromhaemaphysalislongicornisacariixodidaeticks AT renqiaoyun characterizationofanmlphomologuefromhaemaphysalislongicornisacariixodidaeticks AT guanguiquan characterizationofanmlphomologuefromhaemaphysalislongicornisacariixodidaeticks AT zhaobo characterizationofanmlphomologuefromhaemaphysalislongicornisacariixodidaeticks AT yinhong characterizationofanmlphomologuefromhaemaphysalislongicornisacariixodidaeticks AT chenronggui characterizationofanmlphomologuefromhaemaphysalislongicornisacariixodidaeticks AT zhaohongying characterizationofanmlphomologuefromhaemaphysalislongicornisacariixodidaeticks AT luojianxun characterizationofanmlphomologuefromhaemaphysalislongicornisacariixodidaeticks AT lixiangrui characterizationofanmlphomologuefromhaemaphysalislongicornisacariixodidaeticks AT liuguangyuan characterizationofanmlphomologuefromhaemaphysalislongicornisacariixodidaeticks |