RESUMO
Triatoma infestans, the main vector of Chagas disease, is a blood-sucking insect. Flight dispersal of adults is the most important mechanism for reinfestation of houses after insecticide spraying. Flight muscles have two glycerol-3-phosphate dehydrogenase (GPDH) isoforms: GPDH-1 is involved in flight metabolism and GPDH-2 provides lipid precursors. In this study, we explored the profile of GPDH expression in females and males adult flight muscles under light/dark cycle, constant light, and constant dark conditions. Under constant dark conditions, GPDH-1 flight muscles of T. infestans showed a rhythmic pattern of transcription synchronous with a rhythmic profile of activity suggesting regulation by the endogenous circadian clock. Otherwise, the GPDH-2 expression analysis showed no regulation by the endogenous clock, but showed that an external factor, such as the dark/light period, was necessary for synchronization of GPDH-2 transcription and activity.
Assuntos
Relógios Circadianos/genética , Regulação da Expressão Gênica , Glicerolfosfato Desidrogenase/genética , Proteínas de Insetos/genética , Insetos Vetores , Triatoma/genética , Animais , Doença de Chagas/transmissão , Feminino , Voo Animal/fisiologia , Glicerolfosfato Desidrogenase/metabolismo , Humanos , Proteínas de Insetos/metabolismo , Isoenzimas/genética , Isoenzimas/metabolismo , Luz , Masculino , Músculos/enzimologia , Fotoperíodo , Transcrição Gênica , Triatoma/enzimologiaRESUMO
Flight muscles of Triatoma infestans have two glycerol-3-phosphate dehydrogenase (GPDH) isoforms: GPDH-1 is involved in flight metabolism and GPDH-2 provides lipid precursors. Total GPDH activity was greater in the natural population and almost only due to GPDH-1. Different expression and activity observed between GPDH isoforms in the natural population and the first laboratory generation was not detected in the second laboratory generation. This pattern may be caused by gradual adaptation to laboratory nutritional conditions. During development, the expression of GPDH-2 increased with a longer time of intake, which would imply an increment in lipid biosynthesis. The GPDH-1 transcript predominated with respect to that of GPDH-2 in the lower nutritional condition, suggesting the necessity of insects to fly during this nutritional status. The transcriptional pattern showed a delay at 22°C. The isoforms activities and transcript patterns in flight muscles suggest transcriptional adaptation to metabolic requirements originated by alternative splicing.
Assuntos
Voo Animal/fisiologia , Regulação Enzimológica da Expressão Gênica , Glicerolfosfato Desidrogenase/genética , Músculos/fisiologia , RNA/isolamento & purificação , Triatoma/enzimologia , Processamento Alternativo , Animais , Eletroforese em Gel Bidimensional , Feminino , Glicerolfosfato Desidrogenase/metabolismo , Insetos Vetores/enzimologia , Insetos Vetores/crescimento & desenvolvimento , Isoformas de Proteínas , RNA/genética , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo , Triatoma/crescimento & desenvolvimento , Triatoma/patogenicidadeRESUMO
Glycerol-3-phosphate dehydrogenase (GPDH) isozymes are differentially expressed among tissues and during flight development. GPDH-1 is involved in the flight-muscle metabolism and GPDH-2 provides precursors for lipid biosynthesis in many tissues. We have isolated and characterized from Triatoma infestans, a Chagas disease vector, two cDNAs encoding for GPDH-1 and GPDH-2 isozymes. The inferred amino acid sequences showed high identity with other GPDH sequences from flying insects. A GPDH-2 transcript was found in fifth instar nymphs, thoracic muscles, adult gonads, and fat bodies. Both isozymes are present in 30-day-old adult thoracic muscle transcripts, and the pattern of expression differs between sexes. The expression of GPDH-1 begins earlier in females, and GPDH-2 is expressed more abundantly in female adult thoracic muscles than in those from males. This finding is consistent with those of other investigators who showed a higher flight initiation probability in T. infestans females than in males.
Assuntos
Voo Animal/fisiologia , Glicerolfosfato Desidrogenase/biossíntese , Insetos Vetores/enzimologia , Isoenzimas/biossíntese , Triatoma/embriologia , Triatoma/enzimologia , Animais , Doença de Chagas/parasitologia , Vetores de Doenças , Expressão Gênica , Insetos Vetores/crescimento & desenvolvimento , Triatoma/crescimento & desenvolvimentoRESUMO
En el plasma humano pueden encontrarse las isoenzimas ósea, hepática e intestinal de fosfatasa alcalina (EC 3.1.3.1). En el plasma de mujeres embarazadas, durante el último trimestre de gestación puede encontrarse otra isoenzima, la fosfatasa alcalina placentaria. Además, en extractos butanólicos de tejido placentario se ha encontrado una isoenzima unida a membrana, la fosfatasa alcalina placentaria de alto peso molecular. En suero de mujeres embarazadas se ha determinado la actividad de fosfatasa alcalina placentaria soluble pero, hasta el momento, no se ha detectado la presencia de la isoenzima de alto peso molecular. En nuestro laboratorio hemos desarrollado un método que permite la detección de fosfatasa alcalina de alto peso molecular en el pellet de plasma centrifugado a 100.000xg. Utilizando el mencionado método hemos determinado la actividad de fosfatasa alcalina placentaria de alto peso molecular en plasma de mujeres embarazadas durante el último trimestre de gestación
Assuntos
Humanos , Feminino , Gravidez , Fosfatase Alcalina/sangue , Placenta/enzimologia , Gravidez/sangue , Fosfatase Alcalina/isolamento & purificação , Fosfatase Alcalina/metabolismo , Peso Molecular , Terceiro Trimestre da GravidezAssuntos
Humanos , Masculino , Feminino , Fosfatase Alcalina , Isoenzimas , Linfoma/diagnóstico , Biomarcadores Tumorais , Neoplasias Hepáticas/diagnóstico , Neoplasias Hepáticas/secundário , Neoplasias Ósseas/diagnóstico , Neoplasias Ósseas/secundário , Teorema de Bayes , Valor Preditivo dos Testes , Sensibilidade e EspecificidadeRESUMO
En el plasma humano pueden encontrarse las isoenzimas ósea, hepática e intestinal de fosfatasa alcalina (EC 3.1.3.1). En el plasma de mujeres embarazadas, durante el último trimestre de gestación puede encontrarse otra isoenzima, la fosfatasa alcalina placentaria. Además, en extractos butanólicos de tejido placentario se ha encontrado una isoenzima unida a membrana, la fosfatasa alcalina placentaria de alto peso molecular. En suero de mujeres embarazadas se ha determinado la actividad de fosfatasa alcalina placentaria soluble pero, hasta el momento, no se ha detectado la presencia de la isoenzima de alto peso molecular. En nuestro laboratorio hemos desarrollado un método que permite la detección de fosfatasa alcalina de alto peso molecular en el pellet de plasma centrifugado a 100.000xg. Utilizando el mencionado método hemos determinado la actividad de fosfatasa alcalina placentaria de alto peso molecular en plasma de mujeres embarazadas durante el último trimestre de gestación (AU)