Estudo funcional da via PI3K/AKT em Aedes aegypti
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Data
2018-07-23
Autores
Nunes, Tinoco Nunes
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Universidade Estadual Paulista (Unesp)
Resumo
Aedes aegypti é a espécie de mosquito emergente em áreas urbanas com maior impacto na saúde pública, sendo o principal vetor dos arbovírus dengue, Zika e chikungunya. Por esse motivo, se faz indispensável a compreensão de mecanismos moleculares associados a processos fisiológicos em A. aegypti, como resposta imune, comportamento e homeostase intestinal. Conforme observado em outros organismos, a via PI3K/AKT tem papéis importantes no metabolismo, na reprodução, na tolerância ao estresse e na imunidade. A quinase AKT atua como um regulador negativo da via PI3K/AKT, fosforilando o fator de transcrição FOXO e impedindo sua translocação nuclear. Nosso objetivo foi avaliar o perfil transcricional em A. aegypti com o gene akt silenciado e avaliar as consequências deste silenciamento sobre a microbiota bacteriana. Além disso, investigamos uma provável ativação mitocondrial quando do silenciamento de akt. Mostramos que o silenciamento de akt resultou na ativação de genes essenciais para a manutenção da homeostase intestinal do mosquito, como peptídeos antimicrobianos (AMPs) e genes codificadores de enzimas associadas à produção de espécies reativas de oxigênio (ROS). Notavelmente, observou-se uma forte repressão de 11 profenoloxidases (PPOs), além de uma potente indução de genes codificadores de subunidades da enzima NADH desidrogenase, em comparação com o respectivo grupo controle, sugerindo que tal indução esteja associada a um provável aumento da atividade mitocondrial. No contexto comportamental, o transcriptoma de A. aegypti com o gene akt silenciado revelou a modulação de 5 odorant binding proteins (OBPs), das quais todas foram reprimidas no quarto dia após o silenciamento de akt. Além das OBPs, identificamos duas long wavelengh sensitive opsins, ambas reprimidas após dois e quatro dias. Interessantemente, observamos que, dos 345 genes modulados, a grande maioria foi regulada negativamente. O silenciamento de akt, além de modular a carga da microbiota bacteriana de A. aegypti, também modulou táxons específicos após quatro dias de silenciamento, como as Classes Gammaproteobacteria e Betaproteobacteria. Nosso conjunto de dados coloca a via PI3K/AKT no cerce de importantes processos fisiológicos, contribuindo para elucidar mecanismos moleculares até então pouco compreendidos do mosquito.
Aedes aegypti is the emerging mosquito species in urban areas with the greatest impact on public health, being the main vector of arboviruses dengue, Zika and chikungunya. For this reason, it is essential to understand the molecular mechanisms associated with physiological processes in A. aegypti, such as immune response, behavior and intestinal homeostasis. As observed in other organisms, the PI3K / AKT pathway has important roles in metabolism, reproduction, stress tolerance and immunity. AKT kinase acts as a negative regulator of the PI3K / AKT pathway, phosphorylating the FOXO transcription factor and preventing its nuclear translocation. Our objective was to evaluate the transcriptional profile in A. aegypti with the silenced akt gene and to evaluate the consequences of this silencing on the bacterial microbiota. In addition, we investigated a possible mitochondrial activation during the akt silencing. We have shown that akt silencing resulted in the activation of essential genes for the maintenance of mosquito intestinal homeostasis, such as antimicrobial peptides (AMPs) and genes encoding enzymes associated with the production of reactive oxygen species (ROS). Notably, a strong repression of 11 profenoloxidases (PPOs) was observed, as well as a potent induction of genes encoding subunits of the NADH dehydrogenase enzyme, in comparison with the respective control group, suggesting that such induction is associated with a possible increase in mitochondrial activity. In the behavioral context, the A. aegypti transcriptome with the mutated akt gene revealed the modulation of 5 odorant binding proteins (OBPs), all of which were repressed on the fourth day after akt silencing. In addition to the OBPs, we identified two long wavelengh sensitive opsins, both repressed after two and four days. Interestingly, we observed that of the 345 modulated genes, most were down-regulated. The silencing of akt, besides modulating the bacterial microbiota load of A. aegypti, also modulated specific taxa after four days of silencing, such as the Gammaproteobacteria and Betaproteobacteria classes. Our data set puts the PI3K / AKT pathway in the vicinity of important physiological processes, contributing to elucidate the mosquito's poorly understood molecular mechanisms.
Aedes aegypti is the emerging mosquito species in urban areas with the greatest impact on public health, being the main vector of arboviruses dengue, Zika and chikungunya. For this reason, it is essential to understand the molecular mechanisms associated with physiological processes in A. aegypti, such as immune response, behavior and intestinal homeostasis. As observed in other organisms, the PI3K / AKT pathway has important roles in metabolism, reproduction, stress tolerance and immunity. AKT kinase acts as a negative regulator of the PI3K / AKT pathway, phosphorylating the FOXO transcription factor and preventing its nuclear translocation. Our objective was to evaluate the transcriptional profile in A. aegypti with the silenced akt gene and to evaluate the consequences of this silencing on the bacterial microbiota. In addition, we investigated a possible mitochondrial activation during the akt silencing. We have shown that akt silencing resulted in the activation of essential genes for the maintenance of mosquito intestinal homeostasis, such as antimicrobial peptides (AMPs) and genes encoding enzymes associated with the production of reactive oxygen species (ROS). Notably, a strong repression of 11 profenoloxidases (PPOs) was observed, as well as a potent induction of genes encoding subunits of the NADH dehydrogenase enzyme, in comparison with the respective control group, suggesting that such induction is associated with a possible increase in mitochondrial activity. In the behavioral context, the A. aegypti transcriptome with the mutated akt gene revealed the modulation of 5 odorant binding proteins (OBPs), all of which were repressed on the fourth day after akt silencing. In addition to the OBPs, we identified two long wavelengh sensitive opsins, both repressed after two and four days. Interestingly, we observed that of the 345 modulated genes, most were down-regulated. The silencing of akt, besides modulating the bacterial microbiota load of A. aegypti, also modulated specific taxa after four days of silencing, such as the Gammaproteobacteria and Betaproteobacteria classes. Our data set puts the PI3K / AKT pathway in the vicinity of important physiological processes, contributing to elucidate the mosquito's poorly understood molecular mechanisms.
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Palavras-chave
Transcriptoma, Microbioma, Silenciamento gênico, Via PI3K/AKT, RNA de interferência