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The adult females of Aedes aegypti mosquitoes are facultative hematophagous insects but they are unable to feed on blood right after pupae emergence. The maturation process that takes place during the first post-emergence days, hereafter named hematophagic and gonotrophic capacitation, comprises a set of molecular and physiological changes that prepare the females for the first gonotrophic cycle. Notwithstanding, the molecular bases underlying mosquito hematophagic and gonotrophic capacitation remain obscure. Here, we investigated the molecular and biochemical changes in adult Ae. aegypti along the first four days post-emergence, prior to a blood meal. We performed a RNA-Seq analysis of the head and body, comparing male and female gene expression time courses. A total of 811 and 203 genes were differentially expressed, respectively in the body and head, and both body parts showed early, mid, and late female-specific expression profiles. Female-specific up-regulation of genes involved in muscle development and the oxidative phosphorylation pathway were remarkable features observed in the head. Functional assessment of mitochondrial oxygen consumption in heads showed a gradual increase in respiratory capacity and ATP-linked respiration as a consequence of induced mitochondrial biogenesis and content over time. This pattern strongly suggests that boosting oxidative phosphorylation in heads is a required step towards blood sucking habit. Several salivary gland genes, proteases, and genes involved in DNA replication and repair, ribosome biogenesis, and juvenile hormone signaling were up-regulated specifically in the female body, which may reflect the gonotrophic capacitation. This comprehensive description of molecular and biochemical mechanisms of the hematophagic and gonotrophic capacitation in mosquitoes unravels potentially new targets for vector control., Author summary Aedes aegypti is a vector of several human diseases such as Dengue, Zika, Chikungunya, and Yellow Fever. Only the adult females can feed on blood, important for the production of their offspring, and therefore transmitting these pathogens for the human host. Interestingly, right after their emergence as adults, females show a preference of feeding on nectar, and only after three days, they are fully capacitated to perform a blood meal. Understanding the molecular mechanisms that orchestrate this physiological and biochemical switch is the main topic of the present study. The transcriptome analysis of males and females during the first hours post-emergence reveals that the gene expression in females is tightly regulated, showing three time-dependent profiles of expression. There is an increase in genes related to the oxidative phosphorylation pathway in the female head. Assessment of mitochondrial function revealed a gradual increase in respiration linked to ATP synthesis and mitochondrial content. Also, there is an increase in genes related to the muscular function, suggesting a putative role in muscle development and contraction, required, for example, for pumping of blood. The female body showed an increase in salivary gland related genes and serine proteases, probably related to the ingestion and digestion of the blood. Also, the increase in DNA replication, ribosome biogenesis, and juvenile hormone signaling pathways together with changes in cell cycle control suggest the gonotrophic capacitation for the massive synthesis of yolk proteins precursors during vitellogenesis. All these genes were enhanced even before the first blood meal, which reveals insightful information about the biology of anautogenous mosquitoes. Our data showed the molecular machinery that stands the females hematophagy and gonotrophy, leading to potential new targets for vector control.

Background Mosquitoes feed on plant-derived fluids such as nectar and sap and are exposed to bioactive molecules found in this dietary source. However, the role of such molecules on mosquito vectorial capacity is unknown. Weather has been recognized as a major determinant of the spread of dengue, and plants under abiotic stress increase their production of polyphenols. Results Here, we show that including polyphenols in mosquito meals promoted the activation of AMP-dependent protein kinase (AMPK). AMPK positively regulated midgut autophagy leading to a decrease in bacterial proliferation and an increase in vector lifespan. Suppression of AMPK activity resulted in a 6-fold increase in midgut microbiota. Similarly, inhibition of polyphenol-induced autophagy induced an 8-fold increase in bacterial proliferation. Mosquitoes maintained on the polyphenol diet were readily infected by dengue virus. Conclusion The present findings uncover a new direct route by which exacerbation of autophagy through activation of the AMPK pathway leads to a more efficient control of mosquito midgut microbiota and increases the average mosquito lifespan. Our results suggest for the first time that the polyphenol content and availability of the surrounding vegetation may increase the population of mosquitoes prone to infection with arboviruses., Author Summary The dramatic climate changes currently occurring on our planet may likely influence the biology and the distribution of mosquitoes. Aedes aegypti is a major vector of arboviruses. However, females feed on plants for a few days before feeding on blood for the first time. Plants are sessile and cannot move to search for better environmental conditions. In times of extreme temperatures, drought, or UV radiation plants produce polyphenols that allow plants to survive under such extreme conditions. Polyphenol ingestion by mosquitoes enhances the activity of an enzyme named AMP-activated protein kinase (AMPK). Mosquitoes emerging from pupae leave the aquatic environment and are readily colonized by many bacterial strains. Polyphenol-fed mosquitoes display a huge activation of AMPK which enhances autophagy in the midgut cells that increases the capture and killing of midgut bacteria, thus enhancing vector lifespan. Therefore, early meals on polyphenol sources allow for the regulation of vector microbiota. These data may provide the basis of a novel strategy that promotes bacterial proliferation and a shortening of vector lifespan.

BackgroundProtein Tyrosine Phosphatases (PTPs) are enzymes that catalyze phosphotyrosine dephosphorylation and modulate cell differentiation, growth and metabolism. In mammals, PTPs play a key role in the modulation of canonical pathways involved in metabolism and immunity. PTP1B is the prototype member of classical PTPs and a major target for treating human diseases, such as cancer, obesity and diabetes. These signaling enzymes are, hence, targets of a wide array of inhibitors. Anautogenous mosquitoes rely on blood meals to lay eggs and are vectors of the most prevalent human diseases. Identifying the mosquito ortholog of PTP1B and determining its involvement in egg production is, therefore, important in the search for a novel and crucial target for vector control.Methodology/principal findingsWe conducted an analysis to identify the ortholog of mammalian PTP1B in the Aedes aegypti genome. We identified eight genes coding for classical PTPs. In silico structural and functional analyses of proteins coded by such genes revealed that four of these code for catalytically active enzymes. Among the four genes coding for active PTPs, AAEL001919 exhibits the greatest degree of homology with the mammalian PTP1B. Next, we evaluated the role of this enzyme in egg formation. Blood feeding largely affects AAEL001919 expression, especially in the fat body and ovaries. These tissues are critically involved in the synthesis and storage of vitellogenin, the major yolk protein. Including the classical PTP inhibitor sodium orthovanadate or the PTP substrate DiFMUP in the blood meal decreased vitellogenin synthesis and egg production. Similarly, silencing AAEL001919 using RNA interference (RNAi) assays resulted in 30% suppression of egg production.Conclusions/significanceThe data reported herein implicate, for the first time, a gene that codes for a classical PTP in mosquito egg formation. These findings raise the possibility that this class of enzymes may be used as novel targets to block egg formation in mosquitoes.