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2022-07-26T16:45:09Z2022-07-26T16:45:09Z201912010.1016/j.jinsphys.2019.1039730022-1910http://hdl.handle.net/1843/43629In Lutzomyia longipalpis females, which are the main vectors of Leishmania infantum in the Americas, hematophagy is crucial for ovary development. The control of pH in the midgut during blood digestion is important to the functioning of the digestive enzymes, which release amino acids in the luminal compartment that are then transported through the enterocytes to the hemolymph for delivery to the ovary and other organs. In the present work, we investigated transport systems known as LuloPATs that are present in the midgut of L. longipalpis but not in other organs. These transporters achieve symport of amino acids with H+ ions, and one of them (LuloPAT1) is orthologous to a transporter described in Aedes aegypti. According to our results, the transcription levels of LuloPAT1 increased significantly immediately after a blood meal. Based on the variation of the fluorescence of fluorescein with the pH of the medium, we developed a technique that shows the acidification of the cytoplasm of gut cells when amino acids are cotransported with H+ from the lumen into the enterocytes. In our experiments, the midguts of the sandflies were dissected and opened longitudinally so that added amino acids could enter the enterocytes via the lumen (PAT carriers are apical). LuloPAT1 transporters are part of a complex of mechanisms that act synergistically to promote gut alkalinization as soon as blood intake by the vector occurs. In dissected but not longitudinally opened midguts, added amino acids could only enter through the basolateral region of enterocytes. However, alkalinization of the lumen was observed because the entry of some amino acids into the cytoplasm of enterocytes triggers a luminal alkalinization mechanism independent of LuloPATs. These findings provide new perspectives that will enable the characterization of the set of signaling pathways involved in pH regulation within the L. longipalpis midgut.Em fêmeas de Lutzomyia longipalpis, que são os principais vetores de Leishmania infantum nas Américas, a hematofagia é crucial para o desenvolvimento ovariano. O controle do pH no intestino médio durante a digestão do sangue é importante para o funcionamento das enzimas digestivas, que liberam aminoácidos no compartimento luminal que são então transportado através dos enterócitos para a hemolinfa para entrega ao ovário e outros órgãos. No presente trabalho, investigamos sistemas de transporte conhecidos como LuloPATs que estão presentes no intestino médio de L. longipalpis, mas não em outros órgãos. Esses transportadores realizam simporte de aminoácidos com íons H+, e um deles (LuloPAT1) é ortólogo a um transportador descrito em Aedes aegypti. De acordo com nossos resultados, a transcrição os níveis de LuloPAT1 aumentaram significativamente imediatamente após uma refeição de sangue. Com base na variação do fluorescência da fluoresceína com o pH do meio, desenvolvemos uma técnica que mostra a acidificação da no citoplasma das células intestinais quando os aminoácidos são cotransportados com H+ do lúmen para os enterócitos. Dentro Em nossos experimentos, os intestinos médios dos flebotomíneos foram dissecados e abertos longitudinalmente para que fossem adicionados aminoácidos. ácidos podem entrar nos enterócitos através do lúmen (portadores de PAT são apicais). Os transportadores LuloPAT1 fazem parte de um complexo de mecanismos que atuam sinergicamente para promover a alcalinização intestinal assim que a ingestão de sangue pelo ocorre o vetor. Em intestinos dissecados, mas não abertos longitudinalmente, os aminoácidos adicionados só podem entrar através na região basolateral dos enterócitos. No entanto, a alcalinização do lúmen foi observada devido à entrada de alguns aminoácidos no citoplasma dos enterócitos desencadeia um mecanismo de alcalinização luminal independente LuloPATs. Esses achados fornecem novas perspectivas que permitirão a caracterização do conjunto de sinais vias envolvidas na regulação do pH no intestino médio de L. longipalpis.engUniversidade Federal de Minas GeraisUFMGBrasilICB - DEPARTAMENTO DE BIOQUÍMICA E IMUNOLOGIAICB - DEPARTAMENTO DE PARASITOLOGIAJournal of insect physiologyParasitologiaLutzomyia LongipalpisTransportadores de AminoácidosControle de pH do Intestino MédioTransportadores de AminoácidosSimporte H+Lutzomyia longipalpisAmino Acid TransportersH+ SymportMidgut pH ControlAmino Acid CarriersThe role of LuloPAT amino acid/proton symporters in midgut alkalinization in the sandfly Lutzomyia longipalpis (Diptera – Psychodidae)O papel dos simportadores de aminoácidos/prótons LuloPAT na alcalinização do intestino médio no flebotomíneo Lutzomyia longipalpis (Diptera – Psychodidae)info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articlehttps://www.sciencedirect.com/science/article/pii/S0022191019302264Denise Barguil NepomucenoRafaela Magalhães Macedo PaimRicardo Nascimento AraújoMarcos Horacio PereiraGrasielle Caldas Dávila PessoaLeonardo Barbosa KoerichMauricio Roberto Viana Sant'annaNelder de Figueiredo Gontijoapplication/pdfinfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da UFMGinstname:Universidade Federal de Minas Gerais (UFMG)instacron:UFMGORIGINAL2019_The role of lulopat amino acidproton symporters in midgut alkalinization in the sandfly lutzomyia longipalpis (diptera - psychodidae).pdf2019_The role of lulopat amino acidproton symporters in midgut alkalinization in the sandfly lutzomyia longipalpis (diptera - psychodidae).pdfapplication/pdf1162066https://repositorio.ufmg.br/bitstream/1843/43629/2/2019_The%20role%20of%20lulopat%20amino%20acidproton%20symporters%20in%20midgut%20alkalinization%20in%20the%20sandfly%20lutzomyia%20longipalpis%20%28diptera%20-%20psychodidae%29.pdf53c4093e4aff11c20b33b0160a0671d7MD52LICENSELicense.txtLicense.txttext/plain; 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