Initiator and executioner caspases in salivary gland apoptosis of Rhipicephalus haemaphysaloides
Autor(a) principal: | |
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Data de Publicação: | 2020 |
Outros Autores: | , , , , , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Institucional da UFRGS |
Texto Completo: | http://hdl.handle.net/10183/211561 |
Resumo: | Background: Apoptosis is fundamental in maintaining cell balance in multicellular organisms, and caspases play a crucial role in apoptosis pathways. It is reported that apoptosis plays an important role in tick salivary gland degeneration. Several different caspases have been found in ticks, but the interactions between them are currently unknown. Here, we report three new caspases, isolated from the salivary glands of the tick Rhipicephalus haemaphysaloides. Methods: The full-length cDNA of the RhCaspases 7, 8 and 9 genes were obtained by transcriptome, and RhCaspases 7, 8 and 9 were expressed in E. coli; after protein purification and immunization in mice, specific polyclonal antibodies (PcAb) were created in response to the recombinant protein. Reverse-transcription quantitative PCR (RT-qPCR) and western blot were used to detect the existence of RhCaspases 7, 8 and 9 in ticks. TUNEL assays were used to determine the apoptosis level in salivary glands at different feeding times after gene silencing. The interaction between RhCaspases 7, 8 and 9 were identified by co-transfection assays. Results: The transcription of apoptosis-related genes in R. haemaphysaloides salivary glands increased significantly after tick engorgement. Three caspase-like molecules containing conserved caspase domains were identified and named RhCaspases 7, 8 and 9. RhCaspase8 and RhCaspase9 contain a long pro-domain at their N-terminals. An RT-qPCR assay demonstrated that the transcription of these three caspase genes increased significantly during the engorged periods of the tick developmental stages (engorged larval, nymph, and adult female ticks). Transcriptional levels of RhCaspases 7, 8 and 9 in salivary glands increased more significantly than other tissues post-engorgement. RhCaspase9-RNAi treatment significantly inhibited tick feeding. In contrast, knockdown of RhCaspase7 and RhCaspase8 had no influence on tick feeding. Compared to the control group, apoptosis levels were significantly reduced after interfering with RhCaspase 7, 8 and 9 expressions. Co-transfection assays showed RhCaspase7 was cleaved by RhCaspases 8 and 9, demonstrating that RhCaspases 8 and 9 are initiator caspases and RhCaspase7 is an executioner caspase. Conclusions: To the best of our knowledge, this is the first study to identify initiator and executioner caspases in ticks, confirm the interaction among them, and associate caspase activation with tick salivary gland degeneration. |
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Wang, YananHu, ShanmingTuerdi, MayinuerYu, XinmaoZhang, HoushuangZhou, YongzhiCao, JieVaz Junior, Itabajara da SilvaZhou, Jinlin2020-07-09T03:41:12Z20201756-3305http://hdl.handle.net/10183/211561001115235Background: Apoptosis is fundamental in maintaining cell balance in multicellular organisms, and caspases play a crucial role in apoptosis pathways. It is reported that apoptosis plays an important role in tick salivary gland degeneration. Several different caspases have been found in ticks, but the interactions between them are currently unknown. Here, we report three new caspases, isolated from the salivary glands of the tick Rhipicephalus haemaphysaloides. Methods: The full-length cDNA of the RhCaspases 7, 8 and 9 genes were obtained by transcriptome, and RhCaspases 7, 8 and 9 were expressed in E. coli; after protein purification and immunization in mice, specific polyclonal antibodies (PcAb) were created in response to the recombinant protein. Reverse-transcription quantitative PCR (RT-qPCR) and western blot were used to detect the existence of RhCaspases 7, 8 and 9 in ticks. TUNEL assays were used to determine the apoptosis level in salivary glands at different feeding times after gene silencing. The interaction between RhCaspases 7, 8 and 9 were identified by co-transfection assays. Results: The transcription of apoptosis-related genes in R. haemaphysaloides salivary glands increased significantly after tick engorgement. Three caspase-like molecules containing conserved caspase domains were identified and named RhCaspases 7, 8 and 9. RhCaspase8 and RhCaspase9 contain a long pro-domain at their N-terminals. An RT-qPCR assay demonstrated that the transcription of these three caspase genes increased significantly during the engorged periods of the tick developmental stages (engorged larval, nymph, and adult female ticks). Transcriptional levels of RhCaspases 7, 8 and 9 in salivary glands increased more significantly than other tissues post-engorgement. RhCaspase9-RNAi treatment significantly inhibited tick feeding. In contrast, knockdown of RhCaspase7 and RhCaspase8 had no influence on tick feeding. Compared to the control group, apoptosis levels were significantly reduced after interfering with RhCaspase 7, 8 and 9 expressions. Co-transfection assays showed RhCaspase7 was cleaved by RhCaspases 8 and 9, demonstrating that RhCaspases 8 and 9 are initiator caspases and RhCaspase7 is an executioner caspase. Conclusions: To the best of our knowledge, this is the first study to identify initiator and executioner caspases in ticks, confirm the interaction among them, and associate caspase activation with tick salivary gland degeneration.application/pdfengParasites & Vectors. London. Vol. 13 (2020), 288, 15 p.CaspasesApoptoseGlândulas salivaresRhipicephalus haemaphysaloidesTranscriptomaSalivary gland degenerationTranscriptomeApoptosisCaspaseInitiator and executioner caspases in salivary gland apoptosis of Rhipicephalus haemaphysaloidesEstrangeiroinfo:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da UFRGSinstname:Universidade Federal do Rio Grande do Sul (UFRGS)instacron:UFRGSTEXT001115235.pdf.txt001115235.pdf.txtExtracted Texttext/plain57108http://www.lume.ufrgs.br/bitstream/10183/211561/2/001115235.pdf.txt87082db3d903cab803aef99640eff4ceMD52ORIGINAL001115235.pdfTexto completo (inglês)application/pdf2964045http://www.lume.ufrgs.br/bitstream/10183/211561/1/001115235.pdff94b514e6dc8a4250543e97105450801MD5110183/2115612020-07-10 03:41:56.679526oai:www.lume.ufrgs.br:10183/211561Repositório de PublicaçõesPUBhttps://lume.ufrgs.br/oai/requestopendoar:2020-07-10T06:41:56Repositório Institucional da UFRGS - Universidade Federal do Rio Grande do Sul (UFRGS)false |
dc.title.pt_BR.fl_str_mv |
Initiator and executioner caspases in salivary gland apoptosis of Rhipicephalus haemaphysaloides |
title |
Initiator and executioner caspases in salivary gland apoptosis of Rhipicephalus haemaphysaloides |
spellingShingle |
Initiator and executioner caspases in salivary gland apoptosis of Rhipicephalus haemaphysaloides Wang, Yanan Caspases Apoptose Glândulas salivares Rhipicephalus haemaphysaloides Transcriptoma Salivary gland degeneration Transcriptome Apoptosis Caspase |
title_short |
Initiator and executioner caspases in salivary gland apoptosis of Rhipicephalus haemaphysaloides |
title_full |
Initiator and executioner caspases in salivary gland apoptosis of Rhipicephalus haemaphysaloides |
title_fullStr |
Initiator and executioner caspases in salivary gland apoptosis of Rhipicephalus haemaphysaloides |
title_full_unstemmed |
Initiator and executioner caspases in salivary gland apoptosis of Rhipicephalus haemaphysaloides |
title_sort |
Initiator and executioner caspases in salivary gland apoptosis of Rhipicephalus haemaphysaloides |
author |
Wang, Yanan |
author_facet |
Wang, Yanan Hu, Shanming Tuerdi, Mayinuer Yu, Xinmao Zhang, Houshuang Zhou, Yongzhi Cao, Jie Vaz Junior, Itabajara da Silva Zhou, Jinlin |
author_role |
author |
author2 |
Hu, Shanming Tuerdi, Mayinuer Yu, Xinmao Zhang, Houshuang Zhou, Yongzhi Cao, Jie Vaz Junior, Itabajara da Silva Zhou, Jinlin |
author2_role |
author author author author author author author author |
dc.contributor.author.fl_str_mv |
Wang, Yanan Hu, Shanming Tuerdi, Mayinuer Yu, Xinmao Zhang, Houshuang Zhou, Yongzhi Cao, Jie Vaz Junior, Itabajara da Silva Zhou, Jinlin |
dc.subject.por.fl_str_mv |
Caspases Apoptose Glândulas salivares Rhipicephalus haemaphysaloides Transcriptoma |
topic |
Caspases Apoptose Glândulas salivares Rhipicephalus haemaphysaloides Transcriptoma Salivary gland degeneration Transcriptome Apoptosis Caspase |
dc.subject.eng.fl_str_mv |
Salivary gland degeneration Transcriptome Apoptosis Caspase |
description |
Background: Apoptosis is fundamental in maintaining cell balance in multicellular organisms, and caspases play a crucial role in apoptosis pathways. It is reported that apoptosis plays an important role in tick salivary gland degeneration. Several different caspases have been found in ticks, but the interactions between them are currently unknown. Here, we report three new caspases, isolated from the salivary glands of the tick Rhipicephalus haemaphysaloides. Methods: The full-length cDNA of the RhCaspases 7, 8 and 9 genes were obtained by transcriptome, and RhCaspases 7, 8 and 9 were expressed in E. coli; after protein purification and immunization in mice, specific polyclonal antibodies (PcAb) were created in response to the recombinant protein. Reverse-transcription quantitative PCR (RT-qPCR) and western blot were used to detect the existence of RhCaspases 7, 8 and 9 in ticks. TUNEL assays were used to determine the apoptosis level in salivary glands at different feeding times after gene silencing. The interaction between RhCaspases 7, 8 and 9 were identified by co-transfection assays. Results: The transcription of apoptosis-related genes in R. haemaphysaloides salivary glands increased significantly after tick engorgement. Three caspase-like molecules containing conserved caspase domains were identified and named RhCaspases 7, 8 and 9. RhCaspase8 and RhCaspase9 contain a long pro-domain at their N-terminals. An RT-qPCR assay demonstrated that the transcription of these three caspase genes increased significantly during the engorged periods of the tick developmental stages (engorged larval, nymph, and adult female ticks). Transcriptional levels of RhCaspases 7, 8 and 9 in salivary glands increased more significantly than other tissues post-engorgement. RhCaspase9-RNAi treatment significantly inhibited tick feeding. In contrast, knockdown of RhCaspase7 and RhCaspase8 had no influence on tick feeding. Compared to the control group, apoptosis levels were significantly reduced after interfering with RhCaspase 7, 8 and 9 expressions. Co-transfection assays showed RhCaspase7 was cleaved by RhCaspases 8 and 9, demonstrating that RhCaspases 8 and 9 are initiator caspases and RhCaspase7 is an executioner caspase. Conclusions: To the best of our knowledge, this is the first study to identify initiator and executioner caspases in ticks, confirm the interaction among them, and associate caspase activation with tick salivary gland degeneration. |
publishDate |
2020 |
dc.date.accessioned.fl_str_mv |
2020-07-09T03:41:12Z |
dc.date.issued.fl_str_mv |
2020 |
dc.type.driver.fl_str_mv |
Estrangeiro info:eu-repo/semantics/article |
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info:eu-repo/semantics/publishedVersion |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://hdl.handle.net/10183/211561 |
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1756-3305 |
dc.identifier.nrb.pt_BR.fl_str_mv |
001115235 |
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http://hdl.handle.net/10183/211561 |
dc.language.iso.fl_str_mv |
eng |
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eng |
dc.relation.ispartof.pt_BR.fl_str_mv |
Parasites & Vectors. London. Vol. 13 (2020), 288, 15 p. |
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openAccess |
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