Characterisation of Phase I genes from invertebrate detoxification mechanisms: The Mytilus sp. case-study
Autor(a) principal: | |
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Data de Publicação: | 2023 |
Tipo de documento: | Dissertação |
Idioma: | eng |
Título da fonte: | Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
Texto Completo: | http://hdl.handle.net/10362/163524 |
Resumo: | In vertebrates, Cytochrome P450 Family 1 (CYP1) of mixed-function oxidases plays a crucial function in the detoxification of numerous polycyclic aromatic hydrocarbons (PAHs). However, despite cytochromes being relatively well-conserved among eukaryotes, the role of CYPs and other enzymes involved in phase I of detoxification remains obscure and often a matter of controversy. In the case of invertebrates, there have been efforts to characterise the gene and the expression products in organisms pertinent to biomonitoring and toxicity testing proce-dures. The existence of functional homologues in aquatic invertebrates is not universally ac-cepted, particularly in bivalve molluscs, which serve as one of the most significant models for aquatic toxicologists. This thesis aims to contribute to the investigation of phase I detoxification mechanisms in the mussel Mytilus sp. as model, by isolating full coding sequences of CYP1A-Like and Epoxide Hydrolase 4; study the expression of CYP1A-Like and EH4 under toxicolog-ical challenge by B[α]P and associate gene expression with biomarkers and histopathology to determinate B[α]P effects. Following the isolation and sequencing of a CYP1A-like mRNA from Mytilus sp., phylogenetic analyses revealed a similarity to sequences belonging to both the CYP1 and 2 Families. Histology revealed that specimens had Increased histopathological in-juries with increased B[α]P concentrations. When subjected to lower B[α]P concentrations, biomarker activity levels were normal, but their activity decreased when exposed to higher concentrations. When exposed to lower B[α]P concentrations, CYP expression was normal, whereas expression had tendency to diminish when exposed to higher concentrations. This may indicate that either B[α]P in higher concentrations inhibits detoxification mechanisms or that other CYPs may be responsible for detoxification. |
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Characterisation of Phase I genes from invertebrate detoxification mechanisms: The Mytilus sp. case-studyMusselPhase IDetoxificationSequencingBiomarkersB[α]PDomínio/Área Científica::Engenharia e Tecnologia::Outras Engenharias e TecnologiasIn vertebrates, Cytochrome P450 Family 1 (CYP1) of mixed-function oxidases plays a crucial function in the detoxification of numerous polycyclic aromatic hydrocarbons (PAHs). However, despite cytochromes being relatively well-conserved among eukaryotes, the role of CYPs and other enzymes involved in phase I of detoxification remains obscure and often a matter of controversy. In the case of invertebrates, there have been efforts to characterise the gene and the expression products in organisms pertinent to biomonitoring and toxicity testing proce-dures. The existence of functional homologues in aquatic invertebrates is not universally ac-cepted, particularly in bivalve molluscs, which serve as one of the most significant models for aquatic toxicologists. This thesis aims to contribute to the investigation of phase I detoxification mechanisms in the mussel Mytilus sp. as model, by isolating full coding sequences of CYP1A-Like and Epoxide Hydrolase 4; study the expression of CYP1A-Like and EH4 under toxicolog-ical challenge by B[α]P and associate gene expression with biomarkers and histopathology to determinate B[α]P effects. Following the isolation and sequencing of a CYP1A-like mRNA from Mytilus sp., phylogenetic analyses revealed a similarity to sequences belonging to both the CYP1 and 2 Families. Histology revealed that specimens had Increased histopathological in-juries with increased B[α]P concentrations. When subjected to lower B[α]P concentrations, biomarker activity levels were normal, but their activity decreased when exposed to higher concentrations. When exposed to lower B[α]P concentrations, CYP expression was normal, whereas expression had tendency to diminish when exposed to higher concentrations. This may indicate that either B[α]P in higher concentrations inhibits detoxification mechanisms or that other CYPs may be responsible for detoxification.Nos vertebrados, o Citocromo P450, Família 1 (CYP1) desempenha uma função crucial na desintoxicação de numerosos hidrocarbonetos aromáticos policíclicos (HAPs). No entanto, no caso dos invertebrados, tem havido esforços para caracterizar o gene e os produtos de ex-pressão em organismos pertinentes aos procedimentos de bio monitoramento e testes de toxicidade. A existência de homólogos funcionais em invertebrados aquáticos não é univer-salmente aceite, particularmente em moluscos bivalves, que servem como um dos modelos mais significativos para toxicologistas aquáticos. Esta tese pretende contribuir para a investi-gação dos mecanismos de desintoxicação fase I tendo o mexilhão Mytilus sp. como modelo, isolando sequências de codificação completas de CYP1A-Like e Epóxido Hidrolase 4; estudar a expressão de CYP1A-Like e EH4 sob desafio toxicológico por B[α]P e associar a expressão genética com biomarcadores e histopatologia para determinar os efeitos do B[α]P. Após o isolamento e sequenciação do mRNA CYP1A-Like de Mytilus sp., as análises filogenéticas revelaram uma semelhança com sequências pertencentes às famílias CYP1 e 2. A análise histológica revelou que os espécimes apresentavam um aumento de alterações associado com o aumento das concentrações de B[α]P. Quando submetidos a concentrações mais bai-xas de B[α]P, os níveis de atividade dos biomarcadores foram normais, mas sua atividade diminuiu quando expostos a concentrações mais altas. Quando exposto a concentrações mais baixas de B[α]P, a expressão de CYP foi normal, enquanto a expressão teve tendência a diminuir quando exposta a concentrações mais elevadas. Isto pode indicar que o B[α]P em concentrações mais elevadas inibe os mecanismos de desintoxicação ou que outros CYPs podem ser responsáveis pela desintoxicação.Costa, PedroDiniz, MárioRUNVitorino, João Daniel Brás2024-02-14T16:30:37Z2023-102023-10-01T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisapplication/pdfhttp://hdl.handle.net/10362/163524enginfo:eu-repo/semantics/openAccessreponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãoinstacron:RCAAP2024-03-11T05:47:08Zoai:run.unl.pt:10362/163524Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-20T03:59:26.951932Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãofalse |
dc.title.none.fl_str_mv |
Characterisation of Phase I genes from invertebrate detoxification mechanisms: The Mytilus sp. case-study |
title |
Characterisation of Phase I genes from invertebrate detoxification mechanisms: The Mytilus sp. case-study |
spellingShingle |
Characterisation of Phase I genes from invertebrate detoxification mechanisms: The Mytilus sp. case-study Vitorino, João Daniel Brás Mussel Phase I Detoxification Sequencing Biomarkers B[α]P Domínio/Área Científica::Engenharia e Tecnologia::Outras Engenharias e Tecnologias |
title_short |
Characterisation of Phase I genes from invertebrate detoxification mechanisms: The Mytilus sp. case-study |
title_full |
Characterisation of Phase I genes from invertebrate detoxification mechanisms: The Mytilus sp. case-study |
title_fullStr |
Characterisation of Phase I genes from invertebrate detoxification mechanisms: The Mytilus sp. case-study |
title_full_unstemmed |
Characterisation of Phase I genes from invertebrate detoxification mechanisms: The Mytilus sp. case-study |
title_sort |
Characterisation of Phase I genes from invertebrate detoxification mechanisms: The Mytilus sp. case-study |
author |
Vitorino, João Daniel Brás |
author_facet |
Vitorino, João Daniel Brás |
author_role |
author |
dc.contributor.none.fl_str_mv |
Costa, Pedro Diniz, Mário RUN |
dc.contributor.author.fl_str_mv |
Vitorino, João Daniel Brás |
dc.subject.por.fl_str_mv |
Mussel Phase I Detoxification Sequencing Biomarkers B[α]P Domínio/Área Científica::Engenharia e Tecnologia::Outras Engenharias e Tecnologias |
topic |
Mussel Phase I Detoxification Sequencing Biomarkers B[α]P Domínio/Área Científica::Engenharia e Tecnologia::Outras Engenharias e Tecnologias |
description |
In vertebrates, Cytochrome P450 Family 1 (CYP1) of mixed-function oxidases plays a crucial function in the detoxification of numerous polycyclic aromatic hydrocarbons (PAHs). However, despite cytochromes being relatively well-conserved among eukaryotes, the role of CYPs and other enzymes involved in phase I of detoxification remains obscure and often a matter of controversy. In the case of invertebrates, there have been efforts to characterise the gene and the expression products in organisms pertinent to biomonitoring and toxicity testing proce-dures. The existence of functional homologues in aquatic invertebrates is not universally ac-cepted, particularly in bivalve molluscs, which serve as one of the most significant models for aquatic toxicologists. This thesis aims to contribute to the investigation of phase I detoxification mechanisms in the mussel Mytilus sp. as model, by isolating full coding sequences of CYP1A-Like and Epoxide Hydrolase 4; study the expression of CYP1A-Like and EH4 under toxicolog-ical challenge by B[α]P and associate gene expression with biomarkers and histopathology to determinate B[α]P effects. Following the isolation and sequencing of a CYP1A-like mRNA from Mytilus sp., phylogenetic analyses revealed a similarity to sequences belonging to both the CYP1 and 2 Families. Histology revealed that specimens had Increased histopathological in-juries with increased B[α]P concentrations. When subjected to lower B[α]P concentrations, biomarker activity levels were normal, but their activity decreased when exposed to higher concentrations. When exposed to lower B[α]P concentrations, CYP expression was normal, whereas expression had tendency to diminish when exposed to higher concentrations. This may indicate that either B[α]P in higher concentrations inhibits detoxification mechanisms or that other CYPs may be responsible for detoxification. |
publishDate |
2023 |
dc.date.none.fl_str_mv |
2023-10 2023-10-01T00:00:00Z 2024-02-14T16:30:37Z |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/masterThesis |
format |
masterThesis |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://hdl.handle.net/10362/163524 |
url |
http://hdl.handle.net/10362/163524 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
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info:eu-repo/semantics/openAccess |
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openAccess |
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application/pdf |
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Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
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Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação |
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