Análise da sobrevivência, locomoção e ultraestrutura muscular e mitocondrial de Drosophila melanogaster com parkinsonismo induzido por paraquat
Autor(a) principal: | |
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Data de Publicação: | 2020 |
Tipo de documento: | Dissertação |
Idioma: | por |
Título da fonte: | Biblioteca Digital de Teses e Dissertações da PUC_RS |
Texto Completo: | http://tede2.pucrs.br/tede2/handle/tede/9172 |
Resumo: | In humans, Parkinson's disease (PD) is a neurodegenerative process that can occur sporadically, familial or associated with environmental factors, as pesticides exposure like Paraquat (PQ). Fruit fly, Drosophila melanogaster, has been widely used as a biological model for study of familial PD, involving mutants for genes overexpression or null genes, as well as studies of parkinsonism produced by inducing agents, as PQ. Although there are many studies on neuronal effects of PQ in D. melanogaster, there is scarce research on the effects of PQ on the muscular and mitochondrial ultrastructure of D. melanogaster, neither its effects on survival and locomotion parameters, at different ages, in this same biological model. Thus, the goal of present study was to evaluate survival, locomotion and ultrastructural parameters of mitochondrial and muscular fibers, using a 48 hour survival curve; open field test, evaluated by Anymaze software; and transmission electron microscopy (MET) associated to planar morphometry and stereology techniques, in 2 and 15-days-old D. melanogaster, treated with different PQ doses (0 - control, 10, 50, 100, 150 and 200mM), diluted in medium. We observed that PQ has a higher lethality in 15-days-old flies, with 20% mortality in PQ10mM, 60% in PQ50mM and 100% in doses above PQ100mM. In 2-days-old flies, a 20% mortality can be observed, only at PQ150mM dose, and PQ200mM presents as a lethal dose, in both groups. Regarding locomotion, in 15-days-old animals, PQ10mM and PQ50mM doses induce a significant reduction in the following locomotor parameters: distance travelled, average speed and average speed in mobile time (p <0.05), indicating parkinsonian signs of bradykinesia (slow movements). The 2-days-old animals showed no differences in any of the locomotor parameters analyzed, in any PQ doses, when compared to controls. Reductions in survival and locomotion probably induced by acute treatment with PQ are not associated with ultrastructural changes in mitochondrial and muscular, since in all parameters evaluated in MET images (density, percentage of area covered, area and shape coefficient “Shape Z ”), no differences were found regarding to controls, in both doses and ages evaluated (Control and PQ50mM; 2 and 15-days-old). Thus, this study demonstrates that treatment with PQ is more harmful, at least concerning survival and locomotion, in mature individuals (15-days-old) when compared to young individuals (2-days-old). Furthermore, PQ acute treatment did not cause ultrastructural differences in muscle fibers and mitochondria, as previously described in the literature in D. melanogaster mutants with familial PD, as PINK1 and parkin. |
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Xavier, Léder Lealhttp://lattes.cnpq.br/6959317812068630Neves, Paula Fernanda Ribas2020-07-23T17:05:54Z2020-03-11http://tede2.pucrs.br/tede2/handle/tede/9172In humans, Parkinson's disease (PD) is a neurodegenerative process that can occur sporadically, familial or associated with environmental factors, as pesticides exposure like Paraquat (PQ). Fruit fly, Drosophila melanogaster, has been widely used as a biological model for study of familial PD, involving mutants for genes overexpression or null genes, as well as studies of parkinsonism produced by inducing agents, as PQ. Although there are many studies on neuronal effects of PQ in D. melanogaster, there is scarce research on the effects of PQ on the muscular and mitochondrial ultrastructure of D. melanogaster, neither its effects on survival and locomotion parameters, at different ages, in this same biological model. Thus, the goal of present study was to evaluate survival, locomotion and ultrastructural parameters of mitochondrial and muscular fibers, using a 48 hour survival curve; open field test, evaluated by Anymaze software; and transmission electron microscopy (MET) associated to planar morphometry and stereology techniques, in 2 and 15-days-old D. melanogaster, treated with different PQ doses (0 - control, 10, 50, 100, 150 and 200mM), diluted in medium. We observed that PQ has a higher lethality in 15-days-old flies, with 20% mortality in PQ10mM, 60% in PQ50mM and 100% in doses above PQ100mM. In 2-days-old flies, a 20% mortality can be observed, only at PQ150mM dose, and PQ200mM presents as a lethal dose, in both groups. Regarding locomotion, in 15-days-old animals, PQ10mM and PQ50mM doses induce a significant reduction in the following locomotor parameters: distance travelled, average speed and average speed in mobile time (p <0.05), indicating parkinsonian signs of bradykinesia (slow movements). The 2-days-old animals showed no differences in any of the locomotor parameters analyzed, in any PQ doses, when compared to controls. Reductions in survival and locomotion probably induced by acute treatment with PQ are not associated with ultrastructural changes in mitochondrial and muscular, since in all parameters evaluated in MET images (density, percentage of area covered, area and shape coefficient “Shape Z ”), no differences were found regarding to controls, in both doses and ages evaluated (Control and PQ50mM; 2 and 15-days-old). Thus, this study demonstrates that treatment with PQ is more harmful, at least concerning survival and locomotion, in mature individuals (15-days-old) when compared to young individuals (2-days-old). Furthermore, PQ acute treatment did not cause ultrastructural differences in muscle fibers and mitochondria, as previously described in the literature in D. melanogaster mutants with familial PD, as PINK1 and parkin.Em seres humanos, a doença de Parkinson (DP) é um processo neurodegenerativo que pode ocorrer de forma esporádica, familial ou associada à fatores ambientais, como a exposição à agrotóxicos como o Paraquat (PQ). A mosca da fruta, Drosophila melanogaster, tem sido amplamente utilizada como modelo biológico para o estudo da DP familial, envolvendo mutantes com superexpressão de genes ou genes nulos, além de estudos de parkinsonismo produzido por agentes indutores, como o PQ. Embora existam muitos estudos sobre os efeitos neuronais do PQ em D. melanogaster, são escassas as pesquisas sobre efeitos do PQ na ultraestrutura muscular e mitocondrial de D. melanogaster, tampouco seus efeitos em parâmetros de sobrevivência e locomoção, em diferentes idades, neste mesmo modelo biológico. Desse modo, o objetivo do presente trabalho foi avaliar a sobrevivência, locomoção e parâmetros ultraestruturais mitocondriais e musculares, utilizando-se curva de sobrevivência em 48 horas; teste de campo aberto, avaliado pelo software Anymaze; e microscopia eletrônica de transmissão (MET) associada à técnicas de morfometria planar e estereologia, em D. melanogaster de 2 e 15 dias de idade, tratadas com diferentes doses de PQ (0 – controle, 10, 50, 100, 150 e 200mM), diluídas no meio alimentar. Observamos que o PQ apresenta maior letalidade em moscas de 15 dias de idade, apresentando 20% de mortalidade em PQ10mM, 60% em PQ50mM e 100% em doses acima de PQ100mM. Em moscas de 2 dias de idade, uma mortalidade de 20% pode ser observada, apenas na dose de PQ150mM, sendo que PQ200mM apresenta-se como dose letal, em ambos os grupos. Em relação a locomoção, em animais de 15 dias de idade, doses de PQ10mM e PQ50mM induzem significativa redução nos seguintes parâmetros locomotores: distância percorrida, velocidade média e velocidade média no tempo móvel (p <0,05), indicando sinais parkinsonianos de bradicinesia (movimentos lentos). Os animais de 2 dias não apresentaram diferenças em nenhum dos parâmetros locomotores analisados, em nenhuma das doses de PQ, quando comparados aos controles. As reduções de sobrevivência e locomoção provavelmente induzidas pelo tratamento agudo com PQ não estão associados à alterações ultraestruturais mitocondriais e musculares, uma vez que em todos parâmetros avaliados nas imagens de MET (densidade, porcentagem de área coberta, área e coeficiente de forma “Shape Z”), não foram encontradas diferenças em relação aos controles, em ambas as doses e idades avaliadas (Controle e PQ50mM; 2 e 15 dias de idade). Sendo assim, este estudo demonstra que o tratamento com PQ é mais nocivo, ao menos em relação à sobrevivência e a locomoção, em indivíduos maduros (15 dias de idade) quando comparados a indivíduos jovens (2 dias de idade). Além disso, o tratamento agudo com PQ não produz diferenças ultraestruturais em fibras musculares e mitocôndrias, como é descrito na literatura em mutantes de D. melanogaster com DP familial, como PINK1 e parkin.Submitted by PPG Biologia Celular e Molecular (bcm@pucrs.br) on 2020-05-22T20:20:30Z No. of bitstreams: 1 PAULA_FERNANDA_RIBAS_NEVES_DIS.pdf: 2412144 bytes, checksum: 02173963b14255d18bbedc8988261a9d (MD5)Approved for entry into archive by Clarissa Selbach (clarissa.selbach@pucrs.br) on 2020-07-23T17:00:49Z (GMT) No. of bitstreams: 1 PAULA_FERNANDA_RIBAS_NEVES_DIS.pdf: 2412144 bytes, checksum: 02173963b14255d18bbedc8988261a9d (MD5)Made available in DSpace on 2020-07-23T17:05:54Z (GMT). No. of bitstreams: 1 PAULA_FERNANDA_RIBAS_NEVES_DIS.pdf: 2412144 bytes, checksum: 02173963b14255d18bbedc8988261a9d (MD5) Previous issue date: 2020-03-11Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPESapplication/pdfhttp://tede2.pucrs.br:80/tede2/retrieve/178370/DIS_PAULA_FERNANDA_RIBAS_NEVES_CONFIDENCIAL.pdf.jpghttps://tede2.pucrs.br/tede2/retrieve/190335/DIS_PAULA_FERNANDA_RIBAS_NEVES_COMPLETO.pdf.jpgporPontifícia Universidade Católica do Rio Grande do SulPrograma de Pós-Graduação em Biologia Celular e MolecularPUCRSBrasilEscola de CiênciasParkinsonParaquatMitocôndriaMETDrosophila melanogasterParkinsonParaquatMitochondriaTEMDrosophila melanogasterCIENCIAS BIOLOGICAS::BIOLOGIA GERALAnálise da sobrevivência, locomoção e ultraestrutura muscular e mitocondrial de Drosophila melanogaster com parkinsonismo induzido por paraquatinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisTrabalho será publicado como artigo ou livro60 meses23/07/20253463594373552466096500500600-16345593859312446973590462550136975366info:eu-repo/semantics/openAccessreponame:Biblioteca Digital de Teses e Dissertações da PUC_RSinstname:Pontifícia Universidade Católica do Rio Grande do Sul (PUCRS)instacron:PUC_RSORIGINALDIS_PAULA_FERNANDA_RIBAS_NEVES_COMPLETO.pdfDIS_PAULA_FERNANDA_RIBAS_NEVES_COMPLETO.pdfapplication/pdf2412144https://tede2.pucrs.br/tede2/bitstream/tede/9172/5/DIS_PAULA_FERNANDA_RIBAS_NEVES_COMPLETO.pdf02173963b14255d18bbedc8988261a9dMD55THUMBNAILDIS_PAULA_FERNANDA_RIBAS_NEVES_CONFIDENCIAL.pdf.jpgDIS_PAULA_FERNANDA_RIBAS_NEVES_CONFIDENCIAL.pdf.jpgimage/jpeg4096https://tede2.pucrs.br/tede2/bitstream/tede/9172/4/DIS_PAULA_FERNANDA_RIBAS_NEVES_CONFIDENCIAL.pdf.jpgb4fb4da069a388e525c61c3227030d4fMD54DIS_PAULA_FERNANDA_RIBAS_NEVES_COMPLETO.pdf.jpgDIS_PAULA_FERNANDA_RIBAS_NEVES_COMPLETO.pdf.jpgimage/jpeg6236https://tede2.pucrs.br/tede2/bitstream/tede/9172/7/DIS_PAULA_FERNANDA_RIBAS_NEVES_COMPLETO.pdf.jpg56d08888966fe2716a60a60c9b827edaMD57TEXTDIS_PAULA_FERNANDA_RIBAS_NEVES_CONFIDENCIAL.pdf.txtDIS_PAULA_FERNANDA_RIBAS_NEVES_CONFIDENCIAL.pdf.txttext/plain2037https://tede2.pucrs.br/tede2/bitstream/tede/9172/3/DIS_PAULA_FERNANDA_RIBAS_NEVES_CONFIDENCIAL.pdf.txtf9120a493ecd14ef86698956388f8045MD53DIS_PAULA_FERNANDA_RIBAS_NEVES_COMPLETO.pdf.txtDIS_PAULA_FERNANDA_RIBAS_NEVES_COMPLETO.pdf.txttext/plain124880https://tede2.pucrs.br/tede2/bitstream/tede/9172/6/DIS_PAULA_FERNANDA_RIBAS_NEVES_COMPLETO.pdf.txtbfae05bc565481c46177cbab3cf995ffMD56LICENSElicense.txtlicense.txttext/plain; charset=utf-8590https://tede2.pucrs.br/tede2/bitstream/tede/9172/1/license.txt220e11f2d3ba5354f917c7035aadef24MD51tede/91722024-04-26 20:00:15.246oai:tede2.pucrs.br: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Biblioteca Digital de Teses e Dissertaçõeshttp://tede2.pucrs.br/tede2/PRIhttps://tede2.pucrs.br/oai/requestbiblioteca.central@pucrs.br||opendoar:2024-04-26T23:00:15Biblioteca Digital de Teses e Dissertações da PUC_RS - Pontifícia Universidade Católica do Rio Grande do Sul (PUCRS)false |
dc.title.por.fl_str_mv |
Análise da sobrevivência, locomoção e ultraestrutura muscular e mitocondrial de Drosophila melanogaster com parkinsonismo induzido por paraquat |
title |
Análise da sobrevivência, locomoção e ultraestrutura muscular e mitocondrial de Drosophila melanogaster com parkinsonismo induzido por paraquat |
spellingShingle |
Análise da sobrevivência, locomoção e ultraestrutura muscular e mitocondrial de Drosophila melanogaster com parkinsonismo induzido por paraquat Neves, Paula Fernanda Ribas Parkinson Paraquat Mitocôndria MET Drosophila melanogaster Parkinson Paraquat Mitochondria TEM Drosophila melanogaster CIENCIAS BIOLOGICAS::BIOLOGIA GERAL |
title_short |
Análise da sobrevivência, locomoção e ultraestrutura muscular e mitocondrial de Drosophila melanogaster com parkinsonismo induzido por paraquat |
title_full |
Análise da sobrevivência, locomoção e ultraestrutura muscular e mitocondrial de Drosophila melanogaster com parkinsonismo induzido por paraquat |
title_fullStr |
Análise da sobrevivência, locomoção e ultraestrutura muscular e mitocondrial de Drosophila melanogaster com parkinsonismo induzido por paraquat |
title_full_unstemmed |
Análise da sobrevivência, locomoção e ultraestrutura muscular e mitocondrial de Drosophila melanogaster com parkinsonismo induzido por paraquat |
title_sort |
Análise da sobrevivência, locomoção e ultraestrutura muscular e mitocondrial de Drosophila melanogaster com parkinsonismo induzido por paraquat |
author |
Neves, Paula Fernanda Ribas |
author_facet |
Neves, Paula Fernanda Ribas |
author_role |
author |
dc.contributor.advisor1.fl_str_mv |
Xavier, Léder Leal |
dc.contributor.authorLattes.fl_str_mv |
http://lattes.cnpq.br/6959317812068630 |
dc.contributor.author.fl_str_mv |
Neves, Paula Fernanda Ribas |
contributor_str_mv |
Xavier, Léder Leal |
dc.subject.por.fl_str_mv |
Parkinson Paraquat Mitocôndria MET Drosophila melanogaster |
topic |
Parkinson Paraquat Mitocôndria MET Drosophila melanogaster Parkinson Paraquat Mitochondria TEM Drosophila melanogaster CIENCIAS BIOLOGICAS::BIOLOGIA GERAL |
dc.subject.eng.fl_str_mv |
Parkinson Paraquat Mitochondria TEM Drosophila melanogaster |
dc.subject.cnpq.fl_str_mv |
CIENCIAS BIOLOGICAS::BIOLOGIA GERAL |
description |
In humans, Parkinson's disease (PD) is a neurodegenerative process that can occur sporadically, familial or associated with environmental factors, as pesticides exposure like Paraquat (PQ). Fruit fly, Drosophila melanogaster, has been widely used as a biological model for study of familial PD, involving mutants for genes overexpression or null genes, as well as studies of parkinsonism produced by inducing agents, as PQ. Although there are many studies on neuronal effects of PQ in D. melanogaster, there is scarce research on the effects of PQ on the muscular and mitochondrial ultrastructure of D. melanogaster, neither its effects on survival and locomotion parameters, at different ages, in this same biological model. Thus, the goal of present study was to evaluate survival, locomotion and ultrastructural parameters of mitochondrial and muscular fibers, using a 48 hour survival curve; open field test, evaluated by Anymaze software; and transmission electron microscopy (MET) associated to planar morphometry and stereology techniques, in 2 and 15-days-old D. melanogaster, treated with different PQ doses (0 - control, 10, 50, 100, 150 and 200mM), diluted in medium. We observed that PQ has a higher lethality in 15-days-old flies, with 20% mortality in PQ10mM, 60% in PQ50mM and 100% in doses above PQ100mM. In 2-days-old flies, a 20% mortality can be observed, only at PQ150mM dose, and PQ200mM presents as a lethal dose, in both groups. Regarding locomotion, in 15-days-old animals, PQ10mM and PQ50mM doses induce a significant reduction in the following locomotor parameters: distance travelled, average speed and average speed in mobile time (p <0.05), indicating parkinsonian signs of bradykinesia (slow movements). The 2-days-old animals showed no differences in any of the locomotor parameters analyzed, in any PQ doses, when compared to controls. Reductions in survival and locomotion probably induced by acute treatment with PQ are not associated with ultrastructural changes in mitochondrial and muscular, since in all parameters evaluated in MET images (density, percentage of area covered, area and shape coefficient “Shape Z ”), no differences were found regarding to controls, in both doses and ages evaluated (Control and PQ50mM; 2 and 15-days-old). Thus, this study demonstrates that treatment with PQ is more harmful, at least concerning survival and locomotion, in mature individuals (15-days-old) when compared to young individuals (2-days-old). Furthermore, PQ acute treatment did not cause ultrastructural differences in muscle fibers and mitochondria, as previously described in the literature in D. melanogaster mutants with familial PD, as PINK1 and parkin. |
publishDate |
2020 |
dc.date.accessioned.fl_str_mv |
2020-07-23T17:05:54Z |
dc.date.issued.fl_str_mv |
2020-03-11 |
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info:eu-repo/semantics/publishedVersion |
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http://tede2.pucrs.br/tede2/handle/tede/9172 |
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http://tede2.pucrs.br/tede2/handle/tede/9172 |
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por |
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3463594373552466096 |
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Pontifícia Universidade Católica do Rio Grande do Sul |
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Programa de Pós-Graduação em Biologia Celular e Molecular |
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PUCRS |
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Brasil |
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Pontifícia Universidade Católica do Rio Grande do Sul |
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