Compared activity of agonist molecules towards ecdysone receptor in insect cell-based screening system
Autor(a) principal: | |
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Data de Publicação: | 2019 |
Outros Autores: | , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Institucional da UNESP |
Texto Completo: | http://dx.doi.org/10.1590/1808-1657000312019 http://hdl.handle.net/11449/212039 |
Resumo: | The ecdysone receptor, naturally activated by steroidal hormones, is a key protein for molting and reproduction processes of insects. Artificial activation of such receptor by specific pesticides induces an anomalous process of ecdysis, causing death of insects by desiccation and starvation. In this paper, we established a protocol for screening agonistic molecules towards ecdysone receptor of insect cells line S2 (Diptera) and Sf9 (Lepidoptera), transfected with the reporter plasmid ere.b.act.luc. Therefore, we set dose-response curves with the ecdysteroid 20-hydroxyecdysone, the phytoecdysteroid ponasterone-A, and tebufenozide, a pesticide belonging to the class of diacylhydrazines. In both cell lines, the median effective concentration values on reporter gene induction (EC50) of ponasterone-A was the smallest, meaning the most active agonist molecule. In Sf9 cells, tebufenozide had as smaller EC50 than 20-hydroxyecdysone, indicating the high agonistic capability and lepidopteran specificity. The protocol established in this study can be useful for a quick screening and rational research of site-specific pesticides. |
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Compared activity of agonist molecules towards ecdysone receptor in insect cell-based screening systemComparação da atividade de moléculas agonistas em relação ao receptor de ecdisona em um sistema de triagem baseado em linhagens celulares de insetosplasmidtebufenozide20-hydroxyecdysoneponasterone-Aplasmídeotebufenozida20-hidroxiecdisonaponasterona-AThe ecdysone receptor, naturally activated by steroidal hormones, is a key protein for molting and reproduction processes of insects. Artificial activation of such receptor by specific pesticides induces an anomalous process of ecdysis, causing death of insects by desiccation and starvation. In this paper, we established a protocol for screening agonistic molecules towards ecdysone receptor of insect cells line S2 (Diptera) and Sf9 (Lepidoptera), transfected with the reporter plasmid ere.b.act.luc. Therefore, we set dose-response curves with the ecdysteroid 20-hydroxyecdysone, the phytoecdysteroid ponasterone-A, and tebufenozide, a pesticide belonging to the class of diacylhydrazines. In both cell lines, the median effective concentration values on reporter gene induction (EC50) of ponasterone-A was the smallest, meaning the most active agonist molecule. In Sf9 cells, tebufenozide had as smaller EC50 than 20-hydroxyecdysone, indicating the high agonistic capability and lepidopteran specificity. The protocol established in this study can be useful for a quick screening and rational research of site-specific pesticides.O receptor de ecdisona, naturalmente ativado por hormônios esteroidais, é uma proteína-chave nos processos de muda e reprodução de insetos. A ativação artificial desse receptor por meio de pesticidas específicos induz um processo de ecdise anômala, levando o inseto à morte por dessecação e inanição. Neste trabalho, foi estabelecido um protocolo para a triagem de moléculas agonistas em relação ao receptor de ecdisona nas linhagens celulares responsivas S2 (Diptera) e Sf9 (Lepidoptera), transfectadas com o plasmídeo repórter ere.b.act.luc. Para tanto, curvas de dose-resposta foram estabelecidas com o ecdisteroide 20-hidroxiecdisona, o fitoecdisteroide ponasterona-A e tebufenozida, um pesticida pertencente à classe das diacilhidrazinas. Em ambas linhagens celulares, os valores médios de concentração efetiva para indução gênica (EC50) ponasterona-A foram menores, significando que este é o agonista mais potente. Em células Sf9, a tebufenozida apresentou EC50 menor que a 20-hidroxiecdisona, indicando uma alta atividade agonista e especificidade deste inseticida a lepidópteros. O protocolo estabelecido neste trabalho pode ser utilizado para uma rápida triagem e busca racional de pesticidas de alvo bioquímico específico.Universidade Estadual Paulista “Júlio de Mesquita Filho”Universidade Federal de PelotasUniversidade Estadual Paulista “Júlio de Mesquita Filho”Instituto BiológicoUniversidade Estadual Paulista (Unesp)Universidade Federal de PelotasPinto, Ciro Pedro Guidotti [UNESP]Rickes, Letícia NeutzlingZotti, Moisés JoãoGrutzmacher, Anderson Dionei2021-07-14T10:33:33Z2021-07-14T10:33:33Z2019-11-25info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/article-application/pdfhttp://dx.doi.org/10.1590/1808-1657000312019Arquivos do Instituto Biológico. Instituto Biológico, v. 86, p. -, 2019.0020-36531808-1657http://hdl.handle.net/11449/21203910.1590/1808-1657000312019S1808-16572019000100409S1808-16572019000100409.pdfSciELOreponame:Repositório Institucional da UNESPinstname:Universidade Estadual Paulista (UNESP)instacron:UNESPengArquivos do Instituto Biológicoinfo:eu-repo/semantics/openAccess2023-10-31T06:07:44Zoai:repositorio.unesp.br:11449/212039Repositório InstitucionalPUBhttp://repositorio.unesp.br/oai/requestopendoar:29462023-10-31T06:07:44Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP)false |
dc.title.none.fl_str_mv |
Compared activity of agonist molecules towards ecdysone receptor in insect cell-based screening system Comparação da atividade de moléculas agonistas em relação ao receptor de ecdisona em um sistema de triagem baseado em linhagens celulares de insetos |
title |
Compared activity of agonist molecules towards ecdysone receptor in insect cell-based screening system |
spellingShingle |
Compared activity of agonist molecules towards ecdysone receptor in insect cell-based screening system Pinto, Ciro Pedro Guidotti [UNESP] plasmid tebufenozide 20-hydroxyecdysone ponasterone-A plasmídeo tebufenozida 20-hidroxiecdisona ponasterona-A |
title_short |
Compared activity of agonist molecules towards ecdysone receptor in insect cell-based screening system |
title_full |
Compared activity of agonist molecules towards ecdysone receptor in insect cell-based screening system |
title_fullStr |
Compared activity of agonist molecules towards ecdysone receptor in insect cell-based screening system |
title_full_unstemmed |
Compared activity of agonist molecules towards ecdysone receptor in insect cell-based screening system |
title_sort |
Compared activity of agonist molecules towards ecdysone receptor in insect cell-based screening system |
author |
Pinto, Ciro Pedro Guidotti [UNESP] |
author_facet |
Pinto, Ciro Pedro Guidotti [UNESP] Rickes, Letícia Neutzling Zotti, Moisés João Grutzmacher, Anderson Dionei |
author_role |
author |
author2 |
Rickes, Letícia Neutzling Zotti, Moisés João Grutzmacher, Anderson Dionei |
author2_role |
author author author |
dc.contributor.none.fl_str_mv |
Universidade Estadual Paulista (Unesp) Universidade Federal de Pelotas |
dc.contributor.author.fl_str_mv |
Pinto, Ciro Pedro Guidotti [UNESP] Rickes, Letícia Neutzling Zotti, Moisés João Grutzmacher, Anderson Dionei |
dc.subject.por.fl_str_mv |
plasmid tebufenozide 20-hydroxyecdysone ponasterone-A plasmídeo tebufenozida 20-hidroxiecdisona ponasterona-A |
topic |
plasmid tebufenozide 20-hydroxyecdysone ponasterone-A plasmídeo tebufenozida 20-hidroxiecdisona ponasterona-A |
description |
The ecdysone receptor, naturally activated by steroidal hormones, is a key protein for molting and reproduction processes of insects. Artificial activation of such receptor by specific pesticides induces an anomalous process of ecdysis, causing death of insects by desiccation and starvation. In this paper, we established a protocol for screening agonistic molecules towards ecdysone receptor of insect cells line S2 (Diptera) and Sf9 (Lepidoptera), transfected with the reporter plasmid ere.b.act.luc. Therefore, we set dose-response curves with the ecdysteroid 20-hydroxyecdysone, the phytoecdysteroid ponasterone-A, and tebufenozide, a pesticide belonging to the class of diacylhydrazines. In both cell lines, the median effective concentration values on reporter gene induction (EC50) of ponasterone-A was the smallest, meaning the most active agonist molecule. In Sf9 cells, tebufenozide had as smaller EC50 than 20-hydroxyecdysone, indicating the high agonistic capability and lepidopteran specificity. The protocol established in this study can be useful for a quick screening and rational research of site-specific pesticides. |
publishDate |
2019 |
dc.date.none.fl_str_mv |
2019-11-25 2021-07-14T10:33:33Z 2021-07-14T10:33:33Z |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://dx.doi.org/10.1590/1808-1657000312019 Arquivos do Instituto Biológico. Instituto Biológico, v. 86, p. -, 2019. 0020-3653 1808-1657 http://hdl.handle.net/11449/212039 10.1590/1808-1657000312019 S1808-16572019000100409 S1808-16572019000100409.pdf |
url |
http://dx.doi.org/10.1590/1808-1657000312019 http://hdl.handle.net/11449/212039 |
identifier_str_mv |
Arquivos do Instituto Biológico. Instituto Biológico, v. 86, p. -, 2019. 0020-3653 1808-1657 10.1590/1808-1657000312019 S1808-16572019000100409 S1808-16572019000100409.pdf |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
Arquivos do Instituto Biológico |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
- application/pdf |
dc.publisher.none.fl_str_mv |
Instituto Biológico |
publisher.none.fl_str_mv |
Instituto Biológico |
dc.source.none.fl_str_mv |
SciELO reponame:Repositório Institucional da UNESP instname:Universidade Estadual Paulista (UNESP) instacron:UNESP |
instname_str |
Universidade Estadual Paulista (UNESP) |
instacron_str |
UNESP |
institution |
UNESP |
reponame_str |
Repositório Institucional da UNESP |
collection |
Repositório Institucional da UNESP |
repository.name.fl_str_mv |
Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP) |
repository.mail.fl_str_mv |
|
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1803046247839301632 |