RESUMO
BACKGROUND: Obligate blood-feeding insects obtain the nutrients and water necessary to ensure survival from the vertebrate blood. The internal taste sensilla, situated in the pharynx, evaluate the suitability of the ingested food. Here, through multiple approaches, we characterized the pharyngeal organ (PO) of the hematophagous kissing bug Rhodnius prolixus to determine its role in food assessment. The PO, located antero-dorsally in the pharynx, comprises eight taste sensilla that become bathed with the incoming blood. RESULTS: We showed that these taste sensilla house gustatory receptor neurons projecting their axons through the labral nerves to reach the subesophageal zone in the brain. We found that these neurons are electrically activated by relevant appetitive and aversive gustatory stimuli such as NaCl, ATP, and caffeine. Using RNA-Seq, we examined the expression of sensory-related gene families in the PO. We identified gustatory receptors, ionotropic receptors, transient receptor potential channels, pickpocket channels, opsins, takeouts, neuropeptide precursors, neuropeptide receptors, and biogenic amine receptors. RNA interference assays demonstrated that the salt-related pickpocket channel Rproppk014276 is required during feeding of an appetitive solution of NaCl and ATP. CONCLUSIONS: We provide evidence of the role of the pharyngeal organ in food evaluation. This work shows a comprehensive characterization of a pharyngeal taste organ in a hematophagous insect.
Assuntos
Cloreto de Sódio , Paladar , Animais , Paladar/fisiologia , Cloreto de Sódio/farmacologia , Faringe , Insetos , Trifosfato de AdenosinaRESUMO
Methyl farnesoate (MF) plays hormonal regulatory roles in crustaceans. An epoxidated form of MF, known as juvenile hormone (JH), controls metamorphosis and stimulates reproduction in insects. To address the evolutionary significance of MF epoxidation, we generated mosquitoes completely lacking either of the two enzymes that catalyze the last steps of MF/JH biosynthesis and epoxidation, respectively: the JH acid methyltransferase (JHAMT) and the P450 epoxidase CYP15 (EPOX). jhamt-/- larvae lacking both MF and JH died at the onset of metamorphosis. Strikingly, epox-/- mutants, which synthesized MF but no JH, completed the entire life cycle. While epox-/- adults were fertile, the reproductive performance of both sexes was dramatically reduced. Our results suggest that although MF can substitute for the absence of JH in mosquitoes, it is with a significant fitness cost. We propose that MF can fulfill most roles of JH, but its epoxidation to JH was a key innovation providing insects with a reproductive advantage.
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Aedes/genética , Evolução Molecular , Ácidos Graxos Insaturados/metabolismo , Aptidão Genética , Hormônios Juvenis/biossíntese , Aedes/enzimologia , Animais , Feminino , Masculino , Metamorfose Biológica , Reprodução , Sesquiterpenos/metabolismo , Comportamento Sexual AnimalRESUMO
Since the transition from water to land, maintaining water balance has been a key challenge for terrestrial arthropods. We explore factors that allow terrestrial arthropods to survive within a variably dry world and how they shape ecological interactions. Detection of water and hydration is critical for maintaining water content. Efficient regulation of internal water content is accomplished by excretory and osmoregulatory systems that balance water intake and loss. Biochemical and physiological responses are necessary as water content declines to prevent and repair the damage that occurs during dehydration. Desiccation avoidance can occur seasonally or daily via a move to more favorable areas. Dehydration and its avoidance have ecological impacts that extend beyond a single species to alter trophic interactions. As climate changes, evolutionary and ecological processes will be critical to species survival during drought.
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Artrópodes , Animais , Água , Desidratação , Meio Ambiente , SecasRESUMO
Haematophagous mosquitoes need a blood meal to complete their reproductive cycle. To accomplish this, female mosquitoes seek vertebrate hosts, land on them and bite. As their eggs mature, they shift attention away from hosts and towards finding sites to lay eggs. We asked whether females were more tuned to visual cues when a host-related signal, carbon dioxide, was present, and further examined the effect of a blood meal, which shifts behaviour to ovipositing. Using a custom, tethered-flight arena that records wing stroke changes while displaying visual cues, we found the presence of carbon dioxide enhances visual attention towards discrete stimuli and improves contrast sensitivity for host-seeking Aedes aegypti mosquitoes. Conversely, intake of a blood meal reverses vertical bar tracking, a stimulus that non-fed females readily follow. This switch in behaviour suggests that having a blood meal modulates visual attention in mosquitoes, a phenomenon that has been described before in olfaction but not in visually driven behaviours.
Assuntos
Aedes , Animais , Dióxido de Carbono/farmacologia , Sinais (Psicologia) , Comportamento Alimentar , Feminino , OlfatoRESUMO
Transcriptional silencing is a conserved process used by embryonic germ cells to repress somatic fate and maintain totipotency and immortality. In Drosophila, this transcriptional silencing is mediated by polar granule component (pgc). Here, we show that in the adult ovary, pgc is required for timely germline stem cell (GSC) differentiation. Pgc is expressed transiently in the immediate GSC daughter (pre-cystoblast), where it mediates a pulse of transcriptional silencing. This transcriptional silencing mediated by pgc indirectly promotes the accumulation of Cyclin B (CycB) and cell cycle progression into late-G2 phase, when the differentiation factor bag of marbles (bam) is expressed. Pgc mediated accumulation of CycB is also required for heterochromatin deposition, which protects the germ line genome against selfish DNA elements. Our results suggest that transient transcriptional silencing in the pre-cystoblast "re-programs" it away from self-renewal and toward the gamete differentiation program.
Assuntos
Diferenciação Celular/fisiologia , Fase G2/fisiologia , Inativação Gênica/fisiologia , Células Germinativas/metabolismo , Células-Tronco/metabolismo , Animais , Ciclina B/biossíntese , Ciclina B/genética , Proteínas de Drosophila/biossíntese , Proteínas de Drosophila/genética , Drosophila melanogaster , Células Germinativas/citologia , Heterocromatina/genética , Heterocromatina/metabolismo , Células-Tronco/citologiaRESUMO
Female mosquitoes of some species are generalists and will blood-feed on a variety of vertebrate hosts, whereas others display marked host preference. Anopheles gambiae and Aedes aegypti have evolved a strong preference for humans, making them dangerously efficient vectors of malaria and Dengue haemorrhagic fever. Specific host odours probably drive this strong preference because other attractive cues, including body heat and exhaled carbon dioxide (CO2), are common to all warm-blooded hosts. Insects sense odours via several chemosensory receptor families, including the odorant receptors (ORs), membrane proteins that form heteromeric odour-gated ion channels comprising a variable ligand-selective subunit and an obligate co-receptor called Orco (ref. 6). Here we use zinc-finger nucleases to generate targeted mutations in the orco gene of A. aegypti to examine the contribution of Orco and the odorant receptor pathway to mosquito host selection and sensitivity to the insect repellent DEET (N,N-diethyl-meta-toluamide). orco mutant olfactory sensory neurons have greatly reduced spontaneous activity and lack odour-evoked responses. Behaviourally, orco mutant mosquitoes have severely reduced attraction to honey, an odour cue related to floral nectar, and do not respond to human scent in the absence of CO2. However, in the presence of CO2, female orco mutant mosquitoes retain strong attraction to both human and animal hosts, but no longer strongly prefer humans. orco mutant females are attracted to human hosts even in the presence of DEET, but are repelled upon contact, indicating that olfactory- and contact-mediated effects of DEET are mechanistically distinct. We conclude that the odorant receptor pathway is crucial for an anthropophilic vector mosquito to discriminate human from non-human hosts and to be effectively repelled by volatile DEET.
Assuntos
Aedes/genética , Aedes/fisiologia , DEET/farmacologia , Genes de Insetos/genética , Especificidade de Hospedeiro/genética , Repelentes de Insetos/farmacologia , Mutação/genética , Aedes/efeitos dos fármacos , Sequência de Aminoácidos , Animais , Sequência de Bases , DEET/administração & dosagem , Resistência a Medicamentos/efeitos dos fármacos , Feminino , Mel , Especificidade de Hospedeiro/efeitos dos fármacos , Humanos , Repelentes de Insetos/administração & dosagem , Masculino , Dados de Sequência Molecular , Mutagênese Sítio-Dirigida , Neurônios/citologia , Neurônios/efeitos dos fármacos , Odorantes/análise , Condutos Olfatórios/citologia , Condutos Olfatórios/efeitos dos fármacos , VolatilizaçãoRESUMO
BACKGROUND: A complete genome sequence and the advent of genome editing open up non-traditional model organisms to mechanistic genetic studies. The mosquito Aedes aegypti is an important vector of infectious diseases such as dengue, chikungunya, and yellow fever and has a large and complex genome, which has slowed annotation efforts. We used comprehensive transcriptomic analysis of adult gene expression to improve the genome annotation and to provide a detailed tissue-specific catalogue of neural gene expression at different adult behavioral states. RESULTS: We carried out deep RNA sequencing across all major peripheral male and female sensory tissues, the brain and (female) ovary. Furthermore, we examined gene expression across three important phases of the female reproductive cycle, a remarkable example of behavioral switching in which a female mosquito alternates between obtaining blood-meals from humans and laying eggs. Using genome-guided alignments and de novo transcriptome assembly, our re-annotation includes 572 new putative protein-coding genes and updates to 13.5 and 50.3 % of existing transcripts within coding sequences and untranslated regions, respectively. Using this updated annotation, we detail gene expression in each tissue, identifying large numbers of transcripts regulated by blood-feeding and sexually dimorphic transcripts that may provide clues to the biology of male- and female-specific behaviors, such as mating and blood-feeding, which are areas of intensive study for those interested in vector control. CONCLUSIONS: This neurotranscriptome forms a strong foundation for the study of genes in the mosquito nervous system and investigation of sensory-driven behaviors and their regulation. Furthermore, understanding the molecular genetic basis of mosquito chemosensory behavior has important implications for vector control.
Assuntos
Encéfalo/metabolismo , Genoma de Inseto , Ovário/metabolismo , Transcriptoma/genética , Aedes , Animais , Sequência de Bases , Encéfalo/crescimento & desenvolvimento , Feminino , Perfilação da Expressão Gênica , Sequenciamento de Nucleotídeos em Larga Escala/métodos , Masculino , Anotação de Sequência Molecular , Ovário/crescimento & desenvolvimento , FilogeniaRESUMO
Female mosquitoes undergo multiple rounds of reproduction known as gonotrophic cycles (GC). A gonotrophic cycle spans the period from blood meal intake to egg laying. Nutrients from vertebrate host blood are necessary for completing egg development. During oogenesis, a female prepackages mRNA into her oocytes, and these maternal transcripts drive the first 2 h of embryonic development prior to zygotic genome activation. In this study, we profiled transcriptional changes in 1-2 h of Aedes aegypti (Diptera: Culicidae) embryos across 2 GC. We found that homeotic genes which are regulators of embryogenesis are downregulated in embryos from the second gonotrophic cycle. Interestingly, embryos produced by Ae. aegypti females progressively reduced their ability to hatch as the number of GC increased. We show that this fertility decline is due to increased reproductive output and not the mosquitoes' age. Moreover, we found a similar decline in fertility and fecundity across 3 GC in Aedes albopictus. Our results are useful for predicting mosquito population dynamics to inform vector control efforts.
Assuntos
Aedes , Fertilidade , Animais , Aedes/genética , Aedes/fisiologia , Feminino , Fatores Etários , ReproduçãoRESUMO
Where a female mosquito lays her eggs creates the conditions for reproductive success. Here we identify a communal behavior among ovipositing female mosquitoes. When choosing equal breeding sites, gravid Aedes aegypti aggregate more often than expected. This aggregation occurs when water contact is restricted and does not require the presence of eggs. Instead, the aggregation is regulated by the number of females present at the breeding site. Using assays with both occupied and empty oviposition sites, we show that the Orco olfactory co-receptor and a carbon dioxide receptor, Gr3, detect the presence of mosquitoes. orco mutants aggregate more often in empty sites, suggesting attractive olfactory cues influence females to associate with one another. Gr3 mutant females do not prefer either site, suggesting that the CO2 receptor is necessary to evaluate mosquito population density at breeding sites. Further, raising CO2 levels is sufficient to cause wild-type mosquitoes to avoid empty oviposition sites. Our results demonstrate that female mosquitoes can regulate their own population density at breeding sites using attractive and repellent communal cues.
Assuntos
Aedes , Receptores Odorantes , Animais , Feminino , Aedes/genética , Sinais (Psicologia) , Densidade Demográfica , Dióxido de Carbono , OlfatoRESUMO
Survival through periods of drought is critical for mosquitoes to reside in semi-arid regions with humans, but water sources may be limited. Previous studies have shown that dehydrated mosquitoes will increase blood feeding propensity, but how this would occur over extended dry periods is unknown. Following a bloodmeal, prolonged exposure to dry conditions increased secondary blood feeding in mosquitoes by nearly two-fold, and chronic blood feeding allowed mosquitoes to survive twenty days without access to water sources. This refeeding did not alter the number of eggs generated, suggesting this refeeding is for hydration and nutrient replenishment. Exposure to desiccating conditions following a bloodmeal resulted in increased activity, decreased sleep levels, and prompted a return of CO2 sensing before egg deposition. The increased blood feeding during the vitellogenic stage and higher survival during dry periods are predicted to increase pathogen transmission and explain the elevated levels of specific arbovirus cases during dry conditions. These results solidify our understanding of the role of dry periods on mosquito blood feeding and how mosquito dehydration contributes to vectorial capacity and disease transmission dynamics.
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Aedes transmitted arboviral human cases are increasing worldwide and spreading to new areas of the United States of America (USA). These diseases continue to re-emerge likely due to changes in vector ecology, urbanization, human migration, and larger range of climatic suitability. Recent shifts in landscape and weather variables are predicted to impact the habitat patterns of urban mosquitoes such as Aedes aegypti and Aedes albopictus. Miami (FL) is in the tropical zone and an established hotspot for arboviruses, while Charleston (SC) is in the humid subtropical zone and newly vulnerable. Although these coastal cities have distinct climates, both have hot summers. To understand mosquito infestation in both cities and potentiate our surveillance effort, we performed egg collections in the warmest season. We applied remote sensing with land-use cover and weather variation to identify mosquito infestation patterns. Our study found predominant occurrence of Ae. aegypti and, to a lesser extent, Ae. albopictus in both cities. We detected statistically significant positive and negative associations between entomological indicators and most weather variables in combined data from both cities. For all entomological indices, weekly wind speed and relative humidity were significantly positively associated, while precipitation and maximum temperature were significantly negatively associated. Aedes egg abundance was significantly positively associated with open land in Charleston but was negatively associated with vegetation cover in combined data. There is a clear need for further observational studies to determine the impact of climate change on Ae. aegypti and Ae. albopictus infestation in the Southeastern region of the USA.
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The mosquito Aedes aegypti is a prominent vector for arboviruses, but the breadth of mosquito viruses that infects this specie is not fully understood. In the broadest global survey to date of over 200 Ae. aegypti small RNA samples, we detected viral small interfering RNAs (siRNAs) and Piwi interacting RNAs (piRNAs) arising from mosquito viruses. We confirmed that most academic laboratory colonies of Ae. aegypti lack persisting viruses, yet two commercial strains were infected by a novel tombus-like virus. Ae. aegypti from North to South American locations were also teeming with multiple insect viruses, with Anphevirus and a bunyavirus displaying geographical boundaries from the viral small RNA patterns. Asian Ae. aegypti small RNA patterns indicate infections by similar mosquito viruses from the Americas and reveal the first wild example of dengue virus infection generating viral small RNAs. African Ae. aegypti also contained various viral small RNAs including novel viruses only found in these African substrains. Intriguingly, viral long RNA patterns can differ from small RNA patterns, indicative of viral transcripts evading the mosquitoes' RNA interference (RNAi) machinery. To determine whether the viruses we discovered via small RNA sequencing were replicating and transmissible, we infected C6/36 and Aag2 cells with Ae. aegypti homogenates. Through blind passaging, we generated cell lines stably infected by these mosquito viruses which then generated abundant viral siRNAs and piRNAs that resemble the native mosquito viral small RNA patterns. This mosquito small RNA genomics approach augments surveillance approaches for emerging infectious diseases.
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Regenerative processes are critical to maintain tissue homeostasis in high-turnover tissues. At the same time, proliferation of stem and progenitor cells has to be carefully controlled to prevent hyper-proliferative diseases. Mechanisms that ensure this balance, thus promoting proliferative homeostasis, are expected to be critical for longevity in metazoans. The intestinal epithelium of Drosophila provides an accessible model in which to test this prediction. In aging flies, the intestinal epithelium degenerates due to over-proliferation of intestinal stem cells (ISCs) and mis-differentiation of ISC daughter cells, resulting in intestinal dysplasia. Here we show that conditions that impair tissue renewal lead to lifespan shortening, whereas genetic manipulations that improve proliferative homeostasis extend lifespan. These include reduced Insulin/IGF or Jun-N-terminal Kinase (JNK) signaling activities, as well as over-expression of stress-protective genes in somatic stem cell lineages. Interestingly, proliferative activity in aging intestinal epithelia correlates with longevity over a range of genotypes, with maximal lifespan when intestinal proliferation is reduced but not completely inhibited. Our results highlight the importance of the balance between regenerative processes and strategies to prevent hyperproliferative disorders and demonstrate that promoting proliferative homeostasis in aging metazoans is a viable strategy to extend lifespan.
Assuntos
Proliferação de Células , Drosophila melanogaster/crescimento & desenvolvimento , Longevidade , Células-Tronco/citologia , Animais , Animais Geneticamente Modificados , Western Blotting , Senescência Celular , Proteínas de Drosophila/genética , Proteínas de Drosophila/metabolismo , Drosophila melanogaster/citologia , Drosophila melanogaster/genética , Feminino , Regulação da Expressão Gênica no Desenvolvimento , Proteínas de Fluorescência Verde/genética , Proteínas de Fluorescência Verde/metabolismo , Histonas/metabolismo , Homeostase , Proteínas Substratos do Receptor de Insulina , Mucosa Intestinal/metabolismo , Intestinos/citologia , Peptídeos e Proteínas de Sinalização Intracelular/genética , Peptídeos e Proteínas de Sinalização Intracelular/metabolismo , Proteínas Quinases JNK Ativadas por Mitógeno/metabolismo , Masculino , Microscopia Confocal , Mutação , Receptor de Insulina/genética , Receptor de Insulina/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Transdução de Sinais , Células-Tronco/metabolismoRESUMO
Modulation of odorant receptors mRNA induced by prolonged odor exposure is highly correlated with ligand-receptor interactions in Drosophila as well as mammals of the Muridae family. If this response feature is conserved in other organisms, this presents an intriguing initial screening tool when searching for novel receptor-ligand interactions in species with predominantly orphan olfactory receptors. We demonstrate that mRNA modulation in response to 1-octen-3-ol odor exposure occurs in a time- and concentration-dependent manner in Aedes aegypti mosquitoes. To investigate gene expression patterns at a global level, we generated an odor-evoked transcriptome associated with 1-octen-3-ol odor exposure. Transcriptomic data revealed that ORs and OBPs were transcriptionally responsive whereas other chemosensory gene families showed little to no differential expression. Alongside chemosensory gene expression changes, transcriptomic analysis found that prolonged exposure to 1-octen-3-ol modulated xenobiotic response genes, primarily members of the cytochrome P450, insect cuticle proteins, and glucuronosyltransferases families. Together, these findings suggest that mRNA transcriptional modulation of olfactory receptors caused by prolonged odor exposure is pervasive across taxa and can be accompanied by the activation of xenobiotic responses.
Assuntos
Aedes , Receptores Odorantes , Animais , Aedes/fisiologia , Odorantes , Receptores Odorantes/genética , Receptores Odorantes/metabolismo , Transcriptoma , Ligantes , Xenobióticos , RNA Mensageiro , Mamíferos/genéticaRESUMO
Modulation of odorant receptors mRNA induced by prolonged odor exposure is highly correlated with ligand-receptor interactions in Drosophila as well as mammals of the Muridae family. If this response feature is conserved in other organisms, this presents a potentially potent initial screening tool when searching for novel receptor-ligand interactions in species with predominantly orphan olfactory receptors. We demonstrate that mRNA modulation in response to 1-octen-3-ol odor exposure occurs in a time- and concentration-dependent manner in Aedes aegypti mosquitoes. To investigate gene expression patterns at a global level, we generated an odor-evoked transcriptome associated with 1-octen-3-ol odor exposure. Transcriptomic data revealed that ORs and OBPs were transcriptionally responsive whereas other chemosensory gene families showed little to no differential expression. Alongside chemosensory gene expression changes, transcriptomic analysis found that prolonged exposure to 1-octen-3-ol modulated xenobiotic response genes, primarily members of the cytochrome P450, insect cuticle proteins, and glucuronosyltransferases families. Together, these findings suggest that mRNA transcriptional modulation caused by prolonged odor exposure is pervasive across taxa and accompanied by the activation of xenobiotic responses. Furthermore, odor-evoked transcriptomics create a potential screening tool for filtering and identification of chemosensory and xenobiotic targets of interest.
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The uniport olfactometer behavioral assay is currently one of the most reliable single-choice methods to use to study mosquito attraction to olfactory stimuli. It allows for the reproducible calculation of mosquito attraction rate to human hosts or to other olfactory stimuli. Here, we present the design of our modified uniport olfactometer. Consistent carbon-filtered air flows through the assay, creating positive pressure that reduces odor contamination from the room. It includes a precision-milled white acrylic base to facilitate easy setup and consistent placement of the component parts. Our design can be made by a commercial acrylic fabricator or an academic machine shop. This olfactometer is designed to assess the responses of mosquitoes but could be applied to other insects that fly upwind toward an odor stimulus. In a companion protocol, we detail how to perform the experiments with mosquitoes by using the uniport olfactometer.
Assuntos
Culicidae , Odorantes , Animais , HumanosRESUMO
Female mosquitoes respond to the world around them by using chemosensory organs, such as their antennae, to detect volatile compounds emitted from a vertebrate host. These chemosensory systems facilitate the interpretation of external stimuli from the periphery by connecting to the central nervous system and eliciting behaviors necessary for survival, such as obtaining a blood meal. This innate behavior leads to the transmission of pathogens, including dengue virus, chikungunya virus, and Zika virus. Olfaction is a primary sense mosquitoes use to differentiate between vertebrate hosts, and studying it can lead to novel strategies to reduce the risk of disease. In this protocol, we present an olfactory-driven behavioral assay using a uniport olfactometer that measures mosquito attraction rate to a specific stimulus. We include details of the behavioral assay and data analysis as well as how to prepare the mosquitoes before their introduction into the olfactometer. This uniport olfactometer behavioral assay is currently one of the most reliable methods to study mosquito attraction to a single stimulus.
Assuntos
Culicidae , Infecção por Zika virus , Zika virus , Animais , Feminino , Humanos , OlfatoRESUMO
When blood feeding from human hosts, female mosquitoes can transmit life-threatening pathogens to humans, including dengue virus, chikungunya virus, and Zika virus. Olfaction is the primary sense mosquitoes use to locate and differentiate hosts and studying it can lead to new strategies to reduce the risk of disease. To effectively study host-seeking behavior in mosquitoes, a repeatable, quantitative assay that isolates olfaction from other cues is critical for interpreting mosquito behavior. Here, we contribute an overview of methods and best practices for the study of mosquito attraction (or lack thereof) by using olfactometry to quantify behavior. In the accompanying protocols, we present an olfactory-based behavioral assay using a uniport olfactometer that measures mosquito attraction rate to specific stimuli. We include construction details, setup of the uniport olfactometer, details of the behavioral assay, and data analysis guidelines, as well as how to prepare the mosquitoes before their introduction into the olfactometer. This uniport olfactometer behavioral assay is currently one of the most reliable methods to study mosquito attraction to a single olfactory stimulus.
Assuntos
Aedes , Infecção por Zika virus , Zika virus , Animais , Feminino , Humanos , Olfatometria , Olfato , Comportamento AlimentarRESUMO
The mosquito, Aedes aegypti, is highly anthropophilic and transmits debilitating arboviruses within human populations and between humans and non-human primates. Female mosquitoes are attracted to sources of blood by responding to odor plumes that are emitted by their preferred hosts. Acidic volatile compounds, including carboxylic acids, represent particularly salient odors driving this attraction. Importantly, carboxylic acids are major constituents of human sweat and volatiles generated by skin microbes. As such, they are likely to impact human host preference, a dominant factor in disease transmission cycles. A more complete understanding of mosquito host attraction will necessitate the elucidation of molecular mechanisms of volatile odor detection that function in peripheral sensory neurons. Recent studies have shown that members of the variant ionotropic glutamate receptor gene family are necessary for physiological and behavioral responses to acidic volatiles in Aedes. In this study, we have identified a subfamily of variant ionotropic receptors that share sequence homology across several important vector species and are likely to be activated by carboxylic acids. Moreover, we demonstrate that selected members of this subfamily are activated by short-chain carboxylic acids in a heterologous cell expression system. Our results are consistent with the hypothesis that members of this receptor class underlie acidic volatile sensitivity in vector mosquitoes and provide a frame of reference for future development of novel mosquito attractant and repellent technologies.
Assuntos
Aedes , Ácidos Carboxílicos , Animais , Humanos , Feminino , Mosquitos Vetores , Primatas , Odorantes , Aedes/fisiologiaRESUMO
Female Aedes aegypti mosquitoes undergo multiple rounds of reproduction, known as gonotrophic cycles. These cycles span the period from blood meal intake to oviposition. Understanding how reproductive success is maintained across gonotrophic cycles allows for the identification of molecular targets to reduce mosquito population growth. Odorant receptor co-receptor (orco) encodes a conserved insect-specific transmembrane ion channel that complexes with tuning odorant receptors (ORs) to form a functional olfactory receptor. orco expression has been identified in the male and female mosquito germline, but its role is unclear. We report an orco-dependent, maternal effect reduction in fertility after the first gonotrophic cycle. This phenotype was removed by CRISPR-Cas9 reversion of the orco mutant locus. Eggs deposited by orco mutant females are fertilized but the embryos reveal developmental defects, reduced hatching, and changes in ion channel signaling gene transcription. We present an unexpected role for an olfactory receptor pathway in mosquito reproduction.