Epigenetics of marine stickleback: gene expression across a latitudinal gradient

Detalhes bibliográficos
Autor(a) principal: Scheraga, Neal Andrew
Data de Publicação: 2020
Tipo de documento: Dissertação
Idioma: eng
Título da fonte: Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
Texto Completo: http://hdl.handle.net/10400.1/14680
Resumo: Epigenetic mechanisms underlying phenotypic plasticity can be an important factor in the survival of a fish species through a changing climate or in migrating to a new habitat. The threespine stickleback (Gasterosteus aculeatus) is found throughout the northern hemisphere and can adapt via genetic change in relatively few generations to new environments. Epigenetic mechanisms work faster than genetic change, and have the potential to be passed on to future generations, possibly leading to population-wide changes in gene expression and phenotypic variation. To study genes involved in epigenetic mechanisms in stickleback, populations were selected for sampling between Northern Germany and northern Norway. Eleven populations of stickleback were successfully sampled across this latitudinal gradient, and four evenly distributed populations were selected for gene expression analyses in this research. Collected samples were dissected for gonads and pectoral fin muscle (in addition to other organs) and brought to the Alfred Wegener Institute where RNA was extracted. After converting RNA into cDNA, a targeted qPCR approach was performed to test for expression levels of a number of epigenetic actors; DNMT1, DNMT3ab, TET1, TET3, MacroH2A, and Sirtuin2. DNMTs are involved in promoting methylation, TETs actively demethylate cytosine, and MacroH2A and Sirtuin2 are actively involved in cold acclimation. Results from the fieldwork sampling found that stickleback body size (measured as standard length) decreased as latitude increased, in opposition to Bergmann’s rule of species increasing in size toward the poles. Additionally, there was evidence for slight sexual dimorphism in which males were significantly smaller than females across all populations. Furthermore, gravid females were found to be significantly larger than non-gravid females. Results from the target gene qPCR testing found Sirtuin2 to be more expressed in female gonads of northern populations than in southern populations. This is in line with Sirtuin´s role in cold acclimation which would be more beneficial to northern than southern populations. Male gonads showed higher expression of DNMTs and TETs, possibly indicating greater plasticity of epigenetic actors capable of change. This thesis project is the first to study epigenetic differences in fish populations across a latitudinal gradient. Future research could benefit from increasing the sample size (number of individuals and populations) and/or investigating alternative organs that may also show differential gene expression of epigenetic actors. Overall, epigenetic mechanisms are likely to be differentially expressed depending on factors of organ, sex, population, and local environmental factors, all of which can potentially allow greater adaptive potential under climate change.
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spelling Epigenetics of marine stickleback: gene expression across a latitudinal gradientEpigeneticsThreespine sticklebackGene expressionLatitudinal variationDomínio/Área Científica::Ciências Naturais::Outras Ciências NaturaisEpigenetic mechanisms underlying phenotypic plasticity can be an important factor in the survival of a fish species through a changing climate or in migrating to a new habitat. The threespine stickleback (Gasterosteus aculeatus) is found throughout the northern hemisphere and can adapt via genetic change in relatively few generations to new environments. Epigenetic mechanisms work faster than genetic change, and have the potential to be passed on to future generations, possibly leading to population-wide changes in gene expression and phenotypic variation. To study genes involved in epigenetic mechanisms in stickleback, populations were selected for sampling between Northern Germany and northern Norway. Eleven populations of stickleback were successfully sampled across this latitudinal gradient, and four evenly distributed populations were selected for gene expression analyses in this research. Collected samples were dissected for gonads and pectoral fin muscle (in addition to other organs) and brought to the Alfred Wegener Institute where RNA was extracted. After converting RNA into cDNA, a targeted qPCR approach was performed to test for expression levels of a number of epigenetic actors; DNMT1, DNMT3ab, TET1, TET3, MacroH2A, and Sirtuin2. DNMTs are involved in promoting methylation, TETs actively demethylate cytosine, and MacroH2A and Sirtuin2 are actively involved in cold acclimation. Results from the fieldwork sampling found that stickleback body size (measured as standard length) decreased as latitude increased, in opposition to Bergmann’s rule of species increasing in size toward the poles. Additionally, there was evidence for slight sexual dimorphism in which males were significantly smaller than females across all populations. Furthermore, gravid females were found to be significantly larger than non-gravid females. Results from the target gene qPCR testing found Sirtuin2 to be more expressed in female gonads of northern populations than in southern populations. This is in line with Sirtuin´s role in cold acclimation which would be more beneficial to northern than southern populations. Male gonads showed higher expression of DNMTs and TETs, possibly indicating greater plasticity of epigenetic actors capable of change. This thesis project is the first to study epigenetic differences in fish populations across a latitudinal gradient. Future research could benefit from increasing the sample size (number of individuals and populations) and/or investigating alternative organs that may also show differential gene expression of epigenetic actors. Overall, epigenetic mechanisms are likely to be differentially expressed depending on factors of organ, sex, population, and local environmental factors, all of which can potentially allow greater adaptive potential under climate change.O stickleback de três espinhos é um peixe teleósteo encontrado circunglobalmente e é uma espécie modelo em ecologia evolutiva, pois é conhecido por se adaptar em poucas gerações em relação a outras espécies de peixes. Um dos mecanismos usados para se adaptar a ambientes em mudança é através da plasticidade fenotípica por mecanismos epigenéticos. A adição de um grupo metilo à citosina é conhecida como metilação do ADN, e tem o potencial de causar mudanças rápidas em resposta a estímulos ambientais, tais como mudanças na temperatura ou salinidade. A metilação pode ser transmitida aos descendentes, dando a este mecanismo o potencial para produzir mudanças em toda a população de gerações sucessivas. A metilação pode ser criada por metiltransferases de DNA como DNMT1 e DNMT3ab, e removida por proteínas de translocação Ten-eleven como TET1 e TET3. A aclimatação a frio é a capacidade de um organismo de se adaptar a baixas temperaturas, uma característica particularmente útil na sobrevivência em altas latitudes, caracterizadas por temperaturas de água mais baixas. Os genes potencialmente envolvidos na aclimatação a frio incluem MacroH2A e Sirtuin2 que, além desta função, desempenham outras como a modificação da histona e o metabolismo regulador. A aclimatação térmica e a capacidade de um organismo de regular a temperatura é interessante porque cenários futuros de mudança climática prevêem aquecimento, e a capacidade de um organismo para se adaptar a esta mudança pode determinar a sobrevivência de uma população. Para estudar o papel potencial dos mecanismos epigenéticos na adaptação local das populações de três espinhos de stickleback, foi realizado um trabalho de campo para recolher amostras de tecido de stickleback em 11 locais diferentes ao longo das costas do Mar do Norte e da Noruega. A primeira metade do trabalho de campo foi um "loop sul" que ocorreu entre 6 e 28 de Maio de 2019, partindo e regressando ao Instituto Alfred Wegener em List auf Sylt, Alemanha. A rota seguida foi, sucessivamente: de List auf Sylt, Alemanha para Marianger, Dinamarca; Oslofjord, Haugesund, Bergen, Ålesund, Trondheim, Noruega; e finalmente de volta para List auf Sylt, Alemanha. O Stickleback foi capturado com sucesso por rede de cerco em Oslofjord, Haugesund, Bergen e Trondheim na Noruega, por armadilhas do tipo covo em Mariangerfjord, Dinamarca, e por puça em List auf Sylt, Alemanha. A segunda metade do trabalho de campo foi o "loop norte" que ocorreu entre 3 e 20 de junho de 2019, saindo e retornando da Universidade Nord em Bodø, Noruega. A rota geral seguida foi: de Bodø para Vesterålen, Tromsø, Alta, Mo i Rana (NO), e de volta para Bodø. O Stickleback foi capturado com sucesso por armadilhas do tipo covo em Bodø e Vesterålen, e por puça em Alta e Mo i Rana. Por último, foi feita uma curta viagem até Limfjord, Dinamarca, para amostragem de uma população de stickleback com rede de cerco. Finalmente, 305 indivíduos foram capturados, em 11 populações separadas, com cerca de 30 indivíduos capturados em cada local. O stickleback mostrou ser abundante no Atlântico nordeste e apenas dois dos locais, Ålesund e Tromsø, não computaram stickleback. Os resultados da amostragem de stickleback apresentaram uma tendência de diminuição do comprimento padrão com o aumento da latitude. Esta situação é contrária à regra de Bergmann, que afirma que os organismos aumentam de tamanho em altas latitudes. A tendência encontrada pode ser devido a uma variedade de fatores, como regimes térmicos, luz solar, disponibilidade de nutrientes e complexidade do ecossistema. O comprimento do dorso do stickleback diferiu por sexo e mostrou dimorfismo sexual, com os machos sendo consistentemente mais curtos que as fêmeas. Além disso, as fêmeas portadoras de ovos eram maiores do que as fêmeas sem ninhadas desenvolvidas. O comprimento padrão do stickleback de três espinhos foi medido, fotografado e foram dissecados gônadas, músculo peitoral, pele, cérebro e brânquias. O dorso do stickleback foi classificado por gênero somente após a identificação visual de suas gônadas. O stickleback feminino foi designado "gravid female" se seus ovários contivessem óvulos totalmente desenvolvidos e esses óvulos fossem segurados frouxamente ou livremente suspensos na cavidade corporal. Todas as dissecações seguiram o procedimento idêntico de eutanásia por corte preciso da medula espinhal, e amostras de órgãos foram colocadas em tubos Eppendorf contendo RNAlater, rotulados, armazenados em uma criobox, e levados de volta ao Alfred Wegener Institute. No laboratório, as amostras de gônadas e tecido muscular peitoral tiveram o seu RNA extraído utilizando um kit de ADN/RNA da Qiagen® AllPrep™ e as extrações foram normalizadas para conter pelo menos 10 nanogramas por microlitro de RNA. Onze populações tiveram amostras de tecido recuperadas, mas apenas quatro locais foram selecionados para serem usados na análise de expressão gênica para este estudo. Sylt (DE), Bergen, Mo i Rana, e Alta, (NO) foram selecionados porque representavam o comprimento total do gradiente latitudinal desde o Mar do Norte até o extremo norte do Oceano Atlântico. Isso foi feito para aumentar a probabilidade de detecção de diferenças de expressão gênica usando populações o mais afastadas possível.Shama, LisaCastilho, RitaSapientiaScheraga, Neal Andrew2020-09-03T11:32:25Z2020-01-142020-01-14T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisapplication/pdfhttp://hdl.handle.net/10400.1/14680TID:202486109enginfo: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:RCAAP2023-07-24T10:27:01Zoai:sapientia.ualg.pt:10400.1/14680Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-19T20:05:40.829623Repositório 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 Epigenetics of marine stickleback: gene expression across a latitudinal gradient
title Epigenetics of marine stickleback: gene expression across a latitudinal gradient
spellingShingle Epigenetics of marine stickleback: gene expression across a latitudinal gradient
Scheraga, Neal Andrew
Epigenetics
Threespine stickleback
Gene expression
Latitudinal variation
Domínio/Área Científica::Ciências Naturais::Outras Ciências Naturais
title_short Epigenetics of marine stickleback: gene expression across a latitudinal gradient
title_full Epigenetics of marine stickleback: gene expression across a latitudinal gradient
title_fullStr Epigenetics of marine stickleback: gene expression across a latitudinal gradient
title_full_unstemmed Epigenetics of marine stickleback: gene expression across a latitudinal gradient
title_sort Epigenetics of marine stickleback: gene expression across a latitudinal gradient
author Scheraga, Neal Andrew
author_facet Scheraga, Neal Andrew
author_role author
dc.contributor.none.fl_str_mv Shama, Lisa
Castilho, Rita
Sapientia
dc.contributor.author.fl_str_mv Scheraga, Neal Andrew
dc.subject.por.fl_str_mv Epigenetics
Threespine stickleback
Gene expression
Latitudinal variation
Domínio/Área Científica::Ciências Naturais::Outras Ciências Naturais
topic Epigenetics
Threespine stickleback
Gene expression
Latitudinal variation
Domínio/Área Científica::Ciências Naturais::Outras Ciências Naturais
description Epigenetic mechanisms underlying phenotypic plasticity can be an important factor in the survival of a fish species through a changing climate or in migrating to a new habitat. The threespine stickleback (Gasterosteus aculeatus) is found throughout the northern hemisphere and can adapt via genetic change in relatively few generations to new environments. Epigenetic mechanisms work faster than genetic change, and have the potential to be passed on to future generations, possibly leading to population-wide changes in gene expression and phenotypic variation. To study genes involved in epigenetic mechanisms in stickleback, populations were selected for sampling between Northern Germany and northern Norway. Eleven populations of stickleback were successfully sampled across this latitudinal gradient, and four evenly distributed populations were selected for gene expression analyses in this research. Collected samples were dissected for gonads and pectoral fin muscle (in addition to other organs) and brought to the Alfred Wegener Institute where RNA was extracted. After converting RNA into cDNA, a targeted qPCR approach was performed to test for expression levels of a number of epigenetic actors; DNMT1, DNMT3ab, TET1, TET3, MacroH2A, and Sirtuin2. DNMTs are involved in promoting methylation, TETs actively demethylate cytosine, and MacroH2A and Sirtuin2 are actively involved in cold acclimation. Results from the fieldwork sampling found that stickleback body size (measured as standard length) decreased as latitude increased, in opposition to Bergmann’s rule of species increasing in size toward the poles. Additionally, there was evidence for slight sexual dimorphism in which males were significantly smaller than females across all populations. Furthermore, gravid females were found to be significantly larger than non-gravid females. Results from the target gene qPCR testing found Sirtuin2 to be more expressed in female gonads of northern populations than in southern populations. This is in line with Sirtuin´s role in cold acclimation which would be more beneficial to northern than southern populations. Male gonads showed higher expression of DNMTs and TETs, possibly indicating greater plasticity of epigenetic actors capable of change. This thesis project is the first to study epigenetic differences in fish populations across a latitudinal gradient. Future research could benefit from increasing the sample size (number of individuals and populations) and/or investigating alternative organs that may also show differential gene expression of epigenetic actors. Overall, epigenetic mechanisms are likely to be differentially expressed depending on factors of organ, sex, population, and local environmental factors, all of which can potentially allow greater adaptive potential under climate change.
publishDate 2020
dc.date.none.fl_str_mv 2020-09-03T11:32:25Z
2020-01-14
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