Identificação dos mecanismos entomotoxicos de dois compostos inseticidas naturais em modelo experimental alternativo de insetos: extrato contendo anatoxina-a(s) e jaburetox
| Ano de defesa: | 2019 |
|---|---|
| Autor(a) principal: | |
| Orientador(a): | |
| Banca de defesa: | |
| Tipo de documento: | Tese |
| Tipo de acesso: | Acesso aberto |
| dARK ID: | ark:/26339/0013000008ncg |
| Idioma: | por |
| Instituição de defesa: |
Universidade Federal de Santa Maria
Brasil Bioquímica UFSM Programa de Pós-Graduação em Ciências Biológicas: Bioquímica Toxicológica Centro de Ciências Naturais e Exatas |
| Programa de Pós-Graduação: |
Não Informado pela instituição
|
| Departamento: |
Não Informado pela instituição
|
| País: |
Não Informado pela instituição
|
| Palavras-chave em Português: | |
| Link de acesso: | http://repositorio.ufsm.br/handle/1/17545 |
Resumo: | The risk assessment is a stage that aims at the evaluation and characterization of the toxic properties of xenobiotic compounds, conditions of exposure to humans and the environment. The aim of this work was to contribute to the evaluation of the risk potential of natural xenobiotic compounds as Anatoxin-a (s) and Jaburetox, by investigating the mechanisms of entomotoxicity in alternative insect models. In the cerebral homogenates of cockroaches, both extract containing anatoxin-a(s) CECA and trichlorfon (Tn) inhibited acetylcholinesterase activity (AChE). In addition, CECA caused a change in the locomotor pattern of cockroaches characterized by a decrease in distance travelled, increasing the frequency and duration of immobility episodes in a manner similar to that observed with trichlorfon (Tn) organophosphate. CECA also induced an increase in leg grooming activity, but not in antenna grooming activity, similar to the effect of Tn. The CECA and Tn induced negative chronotropism in the insect's heart. It was also found that CECA, Tn, and neostigmine induced a neuromuscular block in vivo preparations of the cockroaches of the species Nauphoeta cinerea. On the other hand, jaburetox (Jbtx), an insecticidal peptide derived from the Canavalia ensiformis urease, has no elucidated mechanism of action, despite interacting with the nervous system of insects. In this work, behavioral, electromyographic and electrophysiological protocols were used to identify the cellular and molecular targets involved in the entomotoxicity of Jbtx in cockroaches and locusts. Jbtx caused a change in the locomotor behavior of the insects characterized by a significant decrease in the distance travelled, followed by an increase in the immobility time in N. cinerea. Jbtx also increased the grooming activity of legs and antennae. Jbtx induced a neuromuscular block in vivo and cardiotoxicity in N. cinerea characterized by the decrease in heart rate of cockroaches. Records of muscle and peripheral nerve potentials of the locusts of the species Locusta migratoria demonstrated that Jbtx increases the amplitude of nerve action potentials without altering the muscle profile. Analysis of voltage clamp in Xenopus laevis heterologously expressed with voltage-dependent sodium channels of Blatella germanica of type BgNaV 1.1, demonstrated that Jbtx increases the amplitude of the sodium currents. The set of unpublished data obtained in this work should contribute in a decisive way improving the effectiveness of the analysis within the environmental risk assessment for the studied compounds, contributing with the optimization of the environmental detection and decontamination processes. Nevertheless, due to the inherent biotechnological context of this work, the toxins studied here may in the near future be used as prototypes for the development of new drugs or pesticides of natural origin. |
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Identificação dos mecanismos entomotoxicos de dois compostos inseticidas naturais em modelo experimental alternativo de insetos: extrato contendo anatoxina-a(s) e jaburetoxIdentification of the entomotoxic mechanisms of two natural insecticides in exterimental alternative insect model: extract containing anatoxin-a (s) and jaburetoxXenobióticosExtrato contendo anatoxina-a(s)JaburetoxNeurotoxicidadeOctopaminaDopaminaDoses subletaisXenobioticsExtract containing anatoxin-a (s)JaburetoxNeurotoxicityOctopamineDopamineSublethal dosesCNPQ::CIENCIAS BIOLOGICAS::BIOQUIMICAThe risk assessment is a stage that aims at the evaluation and characterization of the toxic properties of xenobiotic compounds, conditions of exposure to humans and the environment. The aim of this work was to contribute to the evaluation of the risk potential of natural xenobiotic compounds as Anatoxin-a (s) and Jaburetox, by investigating the mechanisms of entomotoxicity in alternative insect models. In the cerebral homogenates of cockroaches, both extract containing anatoxin-a(s) CECA and trichlorfon (Tn) inhibited acetylcholinesterase activity (AChE). In addition, CECA caused a change in the locomotor pattern of cockroaches characterized by a decrease in distance travelled, increasing the frequency and duration of immobility episodes in a manner similar to that observed with trichlorfon (Tn) organophosphate. CECA also induced an increase in leg grooming activity, but not in antenna grooming activity, similar to the effect of Tn. The CECA and Tn induced negative chronotropism in the insect's heart. It was also found that CECA, Tn, and neostigmine induced a neuromuscular block in vivo preparations of the cockroaches of the species Nauphoeta cinerea. On the other hand, jaburetox (Jbtx), an insecticidal peptide derived from the Canavalia ensiformis urease, has no elucidated mechanism of action, despite interacting with the nervous system of insects. In this work, behavioral, electromyographic and electrophysiological protocols were used to identify the cellular and molecular targets involved in the entomotoxicity of Jbtx in cockroaches and locusts. Jbtx caused a change in the locomotor behavior of the insects characterized by a significant decrease in the distance travelled, followed by an increase in the immobility time in N. cinerea. Jbtx also increased the grooming activity of legs and antennae. Jbtx induced a neuromuscular block in vivo and cardiotoxicity in N. cinerea characterized by the decrease in heart rate of cockroaches. Records of muscle and peripheral nerve potentials of the locusts of the species Locusta migratoria demonstrated that Jbtx increases the amplitude of nerve action potentials without altering the muscle profile. Analysis of voltage clamp in Xenopus laevis heterologously expressed with voltage-dependent sodium channels of Blatella germanica of type BgNaV 1.1, demonstrated that Jbtx increases the amplitude of the sodium currents. The set of unpublished data obtained in this work should contribute in a decisive way improving the effectiveness of the analysis within the environmental risk assessment for the studied compounds, contributing with the optimization of the environmental detection and decontamination processes. Nevertheless, due to the inherent biotechnological context of this work, the toxins studied here may in the near future be used as prototypes for the development of new drugs or pesticides of natural origin.Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPESA avaliação de risco é uma etapa que tem por alvo a avaliação e caracterização das propriedades tóxicas de compostos xenobióticos, condições de exposição a humanos e ao meio ambiente. O objetivo desse trabalho foi contribuir para a avaliação do potencial de risco dos compostos xenobióticos de origem natural, Anatoxina-a(s) e Jaburetox, por meio da investigação dos mecanismos de entomotoxicidade em modelos alternativos de insetos. Nos homogeneizados cerebrais das baratas, tanto o extrato contendo anatoxina-a(s) (CECA), quanto o triclorfon (Tn) inibiu a atividade da acetilcolinesterase (AChE). Além disso, o CECA provocou mudança no padrão locomotor de baratas caracterizado pela diminuição na distância percorrida, aumentando a frequência e a duração dos episódios de imobilidade de maneira similar ao observado com o organofosforado triclorfon (Tn). O CECA também induziu o aumento na atividade de grooming de pernas, mas não na atividade de grooming de antenas, similarmente ao efeito do Tn. O CECA e Tn induziram cronotropismo negativo no coração do inseto. Também foi constatado que o CECA, Tn e neostigmina induziram bloqueio neuromuscular em preparações in vivo da barata da espécie Nauphoeta cinerea. Por outro lado, o jaburetox (Jbtx), um peptídeo inseticida derivado da urease de Canavalia ensiformis, não possui mecanismo de ação elucidado, apesar de interagir com o sistema nervoso de insetos. Nesse trabalho foram empregados protocolos comportamentais, eletromiográficos e eletrofisiológicos para identificar os alvos celulares e moleculares envolvidos na entomotoxicidade do Jbtx em baratas e gafanhotos. O Jbtx causou alteração no comportamento locomotor dos insetos caracterizado por uma diminuição significativa na distância percorrida, seguida por um aumento no tempo de imobilidade em N. cinerea. O Jbtx aumentou a atividade de grooming de pernas e antenas. Jbtx também induziu bloqueio neuromuscular in vivo e cardiotoxicidade em N. cinerea caracterizado pela diminuição da frequência cardíaca das baratas. Registros dos potenciais de ação do músculo e do nervo periférico de gafanhotos da espécie Locusta migratoria demonstraram que o Jbtx aumenta a amplitude dos potenciais de ação do nervo, sem alterar o perfil muscular. Análises de voltage clamp em Xenopus laevis heterologamente expressados com canais de sódio voltagem-dependentes de Blatella germanica do tipo BgNaV 1.1, demonstraram que o Jbtx aumenta a amplitude das correntes de sódio. O conjunto de dados inéditos obtidos nesse trabalho deverá contribuir de maneira decisiva melhorando a efetividade da análise dentro da avaliação de risco ambiental para os compostos estudados, contribuindo com a otimização dos processos de detecção e descontaminação ambiental. Não obstante, devido ao inerente contexto biotecnológico desse trabalho, as toxinas aqui estudadas poderão em um futuro próximo serem empregadas como protótipos para o desenvolvimento de novas drogas ou pesticidas de origem natural.Universidade Federal de Santa MariaBrasilBioquímicaUFSMPrograma de Pós-Graduação em Ciências Biológicas: Bioquímica ToxicológicaCentro de Ciências Naturais e ExatasDal Belo, Cháriston Andréhttp://lattes.cnpq.br/3500651347964867Ávila, Daiana Silva dehttp://lattes.cnpq.br/4355211015887363Silva, Sidnei Moura ehttp://lattes.cnpq.br/3505301292906591Santos, Douglas Silva dos2019-07-24T13:21:16Z2019-07-24T13:21:16Z2019-03-29info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/doctoralThesisapplication/pdfhttp://repositorio.ufsm.br/handle/1/17545ark:/26339/0013000008ncgporAttribution-NonCommercial-NoDerivatives 4.0 Internationalinfo:eu-repo/semantics/openAccessreponame:Manancial - Repositório Digital da UFSMinstname:Universidade Federal de Santa Maria (UFSM)instacron:UFSM2019-07-25T06:02:10Zoai:repositorio.ufsm.br:1/17545Biblioteca Digital de Teses e Dissertaçõeshttps://repositorio.ufsm.br/PUBhttps://repositorio.ufsm.br/oai/requestatendimento.sib@ufsm.br||tedebc@gmail.com||manancial@ufsm.bropendoar:2019-07-25T06:02:10Manancial - Repositório Digital da UFSM - Universidade Federal de Santa Maria (UFSM)false |
| dc.title.none.fl_str_mv |
Identificação dos mecanismos entomotoxicos de dois compostos inseticidas naturais em modelo experimental alternativo de insetos: extrato contendo anatoxina-a(s) e jaburetox Identification of the entomotoxic mechanisms of two natural insecticides in exterimental alternative insect model: extract containing anatoxin-a (s) and jaburetox |
| title |
Identificação dos mecanismos entomotoxicos de dois compostos inseticidas naturais em modelo experimental alternativo de insetos: extrato contendo anatoxina-a(s) e jaburetox |
| spellingShingle |
Identificação dos mecanismos entomotoxicos de dois compostos inseticidas naturais em modelo experimental alternativo de insetos: extrato contendo anatoxina-a(s) e jaburetox Santos, Douglas Silva dos Xenobióticos Extrato contendo anatoxina-a(s) Jaburetox Neurotoxicidade Octopamina Dopamina Doses subletais Xenobiotics Extract containing anatoxin-a (s) Jaburetox Neurotoxicity Octopamine Dopamine Sublethal doses CNPQ::CIENCIAS BIOLOGICAS::BIOQUIMICA |
| title_short |
Identificação dos mecanismos entomotoxicos de dois compostos inseticidas naturais em modelo experimental alternativo de insetos: extrato contendo anatoxina-a(s) e jaburetox |
| title_full |
Identificação dos mecanismos entomotoxicos de dois compostos inseticidas naturais em modelo experimental alternativo de insetos: extrato contendo anatoxina-a(s) e jaburetox |
| title_fullStr |
Identificação dos mecanismos entomotoxicos de dois compostos inseticidas naturais em modelo experimental alternativo de insetos: extrato contendo anatoxina-a(s) e jaburetox |
| title_full_unstemmed |
Identificação dos mecanismos entomotoxicos de dois compostos inseticidas naturais em modelo experimental alternativo de insetos: extrato contendo anatoxina-a(s) e jaburetox |
| title_sort |
Identificação dos mecanismos entomotoxicos de dois compostos inseticidas naturais em modelo experimental alternativo de insetos: extrato contendo anatoxina-a(s) e jaburetox |
| author |
Santos, Douglas Silva dos |
| author_facet |
Santos, Douglas Silva dos |
| author_role |
author |
| dc.contributor.none.fl_str_mv |
Dal Belo, Cháriston André http://lattes.cnpq.br/3500651347964867 Ávila, Daiana Silva de http://lattes.cnpq.br/4355211015887363 Silva, Sidnei Moura e http://lattes.cnpq.br/3505301292906591 |
| dc.contributor.author.fl_str_mv |
Santos, Douglas Silva dos |
| dc.subject.por.fl_str_mv |
Xenobióticos Extrato contendo anatoxina-a(s) Jaburetox Neurotoxicidade Octopamina Dopamina Doses subletais Xenobiotics Extract containing anatoxin-a (s) Jaburetox Neurotoxicity Octopamine Dopamine Sublethal doses CNPQ::CIENCIAS BIOLOGICAS::BIOQUIMICA |
| topic |
Xenobióticos Extrato contendo anatoxina-a(s) Jaburetox Neurotoxicidade Octopamina Dopamina Doses subletais Xenobiotics Extract containing anatoxin-a (s) Jaburetox Neurotoxicity Octopamine Dopamine Sublethal doses CNPQ::CIENCIAS BIOLOGICAS::BIOQUIMICA |
| description |
The risk assessment is a stage that aims at the evaluation and characterization of the toxic properties of xenobiotic compounds, conditions of exposure to humans and the environment. The aim of this work was to contribute to the evaluation of the risk potential of natural xenobiotic compounds as Anatoxin-a (s) and Jaburetox, by investigating the mechanisms of entomotoxicity in alternative insect models. In the cerebral homogenates of cockroaches, both extract containing anatoxin-a(s) CECA and trichlorfon (Tn) inhibited acetylcholinesterase activity (AChE). In addition, CECA caused a change in the locomotor pattern of cockroaches characterized by a decrease in distance travelled, increasing the frequency and duration of immobility episodes in a manner similar to that observed with trichlorfon (Tn) organophosphate. CECA also induced an increase in leg grooming activity, but not in antenna grooming activity, similar to the effect of Tn. The CECA and Tn induced negative chronotropism in the insect's heart. It was also found that CECA, Tn, and neostigmine induced a neuromuscular block in vivo preparations of the cockroaches of the species Nauphoeta cinerea. On the other hand, jaburetox (Jbtx), an insecticidal peptide derived from the Canavalia ensiformis urease, has no elucidated mechanism of action, despite interacting with the nervous system of insects. In this work, behavioral, electromyographic and electrophysiological protocols were used to identify the cellular and molecular targets involved in the entomotoxicity of Jbtx in cockroaches and locusts. Jbtx caused a change in the locomotor behavior of the insects characterized by a significant decrease in the distance travelled, followed by an increase in the immobility time in N. cinerea. Jbtx also increased the grooming activity of legs and antennae. Jbtx induced a neuromuscular block in vivo and cardiotoxicity in N. cinerea characterized by the decrease in heart rate of cockroaches. Records of muscle and peripheral nerve potentials of the locusts of the species Locusta migratoria demonstrated that Jbtx increases the amplitude of nerve action potentials without altering the muscle profile. Analysis of voltage clamp in Xenopus laevis heterologously expressed with voltage-dependent sodium channels of Blatella germanica of type BgNaV 1.1, demonstrated that Jbtx increases the amplitude of the sodium currents. The set of unpublished data obtained in this work should contribute in a decisive way improving the effectiveness of the analysis within the environmental risk assessment for the studied compounds, contributing with the optimization of the environmental detection and decontamination processes. Nevertheless, due to the inherent biotechnological context of this work, the toxins studied here may in the near future be used as prototypes for the development of new drugs or pesticides of natural origin. |
| publishDate |
2019 |
| dc.date.none.fl_str_mv |
2019-07-24T13:21:16Z 2019-07-24T13:21:16Z 2019-03-29 |
| dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
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info:eu-repo/semantics/doctoralThesis |
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doctoralThesis |
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publishedVersion |
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http://repositorio.ufsm.br/handle/1/17545 |
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ark:/26339/0013000008ncg |
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http://repositorio.ufsm.br/handle/1/17545 |
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ark:/26339/0013000008ncg |
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por |
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Attribution-NonCommercial-NoDerivatives 4.0 International info:eu-repo/semantics/openAccess |
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Attribution-NonCommercial-NoDerivatives 4.0 International |
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openAccess |
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application/pdf |
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Universidade Federal de Santa Maria Brasil Bioquímica UFSM Programa de Pós-Graduação em Ciências Biológicas: Bioquímica Toxicológica Centro de Ciências Naturais e Exatas |
| publisher.none.fl_str_mv |
Universidade Federal de Santa Maria Brasil Bioquímica UFSM Programa de Pós-Graduação em Ciências Biológicas: Bioquímica Toxicológica Centro de Ciências Naturais e Exatas |
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reponame:Manancial - Repositório Digital da UFSM instname:Universidade Federal de Santa Maria (UFSM) instacron:UFSM |
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Universidade Federal de Santa Maria (UFSM) |
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UFSM |
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UFSM |
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Manancial - Repositório Digital da UFSM |
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Manancial - Repositório Digital da UFSM - Universidade Federal de Santa Maria (UFSM) |
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atendimento.sib@ufsm.br||tedebc@gmail.com||manancial@ufsm.br |
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1847153368691113984 |