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1.
Arch Insect Biochem Physiol ; 115(4): e22106, 2024 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-38597092

RESUMO

Kissing bugs do not respond to host cues when recently molted and only exhibit robust host-seeking several days after ecdysis. Behavioral plasticity has peripheral correlates in antennal gene expression changes through the week after ecdysis. The mechanisms regulating these peripheral changes are still unknown, but neuropeptide, G-protein coupled receptor, nuclear receptor, and takeout genes likely modulate peripheral sensory physiology. We evaluated their expression in antennal transcriptomes along the first week postecdysis of Rhodnius prolixus 5th instar larvae. Besides, we performed clustering and co-expression analyses to reveal relationships between neuromodulatory (NM) and sensory genes. Significant changes in transcript abundance were detected for 50 NM genes. We identified 73 sensory-related and NM genes that were assigned to nine clusters. According to their expression patterns, clusters were classified into four groups: two including genes up or downregulated immediately after ecdysis; and two with genes with expression altered at day 2. Several NM genes together with sensory genes belong to the first group, suggesting functional interactions. Co-expression network analysis revealed a set of genes that seem to connect with sensory system maturation. Significant expression changes in NM components were described in the antennae of R. prolixus after ecdysis, suggesting that a local NM system acts on antennal physiology. These changes may modify the sensitivity of kissing bugs to host cues during this maturation interval.


Assuntos
Neuropeptídeos , Rhodnius , Triatoma , Animais , Rhodnius/genética , Rhodnius/metabolismo , Neuropeptídeos/genética , Neuropeptídeos/metabolismo , Transcriptoma , Muda
2.
BMC Biol ; 22(1): 63, 2024 Mar 13.
Artigo em Inglês | MEDLINE | ID: mdl-38481317

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 Adenosina
3.
PLoS One ; 18(3): e0282490, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-36867641

RESUMO

BACKGROUND: Rhodnius prolixus is a vector of Chagas disease and has become a model organism to study physiology, behavior, and pathogen interaction. The publication of its genome allowed initiating a process of comparative characterization of the gene expression profiles of diverse organs exposed to varying conditions. Brain processes control the expression of behavior and, as such, mediate immediate adjustment to a changing environment, allowing organisms to maximize their chances to survive and reproduce. The expression of fundamental behavioral processes like feeding requires fine control in triatomines because they obtain their blood meals from potential predators. Therefore, the characterization of gene expression profiles of key components modulating behavior in brain processes, like those of neuropeptide precursors and their receptors, seems fundamental. Here we study global gene expression profiles in the brain of starved R. prolixus fifth instar nymphs by means of RNA sequencing (RNA-Seq). RESULTS: The expression of neuromodulatory genes such as those of precursors of neuropeptides, neurohormones, and their receptors; as well as the enzymes involved in the biosynthesis and processing of neuropeptides and biogenic amines were fully characterized. Other important gene targets such as neurotransmitter receptors, nuclear receptors, clock genes, sensory receptors, and takeouts genes were identified and their gene expression analyzed. CONCLUSION: We propose that the set of neuromodulatory-related genes highly expressed in the brain of starved R. prolixus nymphs deserves functional characterization to allow the subsequent development of tools targeting them for bug control. As the brain is a complex structure that presents functionally specialized areas, future studies should focus on characterizing gene expression profiles in target areas, e.g. mushroom bodies, to complement our current knowledge.


Assuntos
Doença de Chagas , Rhodnius , Animais , Encéfalo , Conhecimento , Ninfa , Expressão Gênica
4.
BMC Genomics ; 23(1): 861, 2022 Dec 30.
Artigo em Inglês | MEDLINE | ID: mdl-36585617

RESUMO

BACKGROUND: Triatoma infestans is the main vector of Chagas disease in the Americas, currently transmitting it in Argentina, Paraguay, and Bolivia. Many T. infestans populations present insecticide resistance, reducing the efficiency of control campaigns. Alternative vector control methods are needed, and molecular targets mediating fundamental physiological processes can be a promising option to manipulate kissing bug behavior. Therefore, it is necessary to characterize the main sensory targets, as well as to determine whether they are modulated by physiological factors. In order to identify gene candidates potentially mediating host cue detection, the antennal transcripts of T. infestans fifth instar larvae were sequenced and assembled. Besides, we evaluated whether a blood meal had an effect on transcriptional profiles, as responsiveness to host-emitted sensory cues depends on bug starvation. RESULTS: The sensory-related gene families of T. infestans were annotated (127 odorant receptors, 38 ionotropic receptors, 11 gustatory receptors, 41 odorant binding proteins, and 25 chemosensory proteins, among others) and compared to those of several other hemipterans, including four triatomine species. Several triatomine-specific lineages representing sensory adaptations developed through the evolution of these blood-feeding heteropterans were identified. As well, we report here various conserved sensory gene orthogroups shared by heteropterans. The absence of the thermosensor pyrexia, of pickpocket receptor subfamilies IV and VII, together with clearly expanded takeout repertoires, are revealed features of the molecular bases of heteropteran antennal physiology. Finally, out of 2,122 genes whose antennal expression was significantly altered by the ingestion of a blood meal, a set of 41 T. infestans sensory-related genes (9 up-regulated; 32 down-regulated) was detected. CONCLUSIONS: We propose that the set of genes presenting nutritionally-triggered modulation on their expression represent candidates to mediate triatomine host-seeking behavior. Besides, the triatomine-specific gene lineages found represent molecular adaptations to their risky natural history that involves stealing blood from an enormously diverse set of vertebrates. Heteropteran gene orthogroups identified may represent unknown features of the sensory specificities of this largest group of hemipteroids. Our work is the first molecular characterization of the peripheral modulation of sensory processes in a non-dipteran vector of human disease.


Assuntos
Doença de Chagas , Triatoma , Animais , Humanos , Triatoma/genética , Triatoma/metabolismo , Transcriptoma , Bolívia , Resistência a Inseticidas
5.
PLoS Negl Trop Dis ; 16(6): e0010060, 2022 06.
Artigo em Inglês | MEDLINE | ID: mdl-35767570

RESUMO

BACKGROUND: Triatoma infestans is the main vector of Chagas disease in the Southern Cone. The resistance to pyrethroid insecticides developed by populations of this species impairs the effectiveness of vector control campaigns in wide regions of Argentina. The study of the global transcriptomic response to pyrethroid insecticides is important to deepen the knowledge about detoxification in triatomines. METHODOLOGY AND FINDINGS: We used RNA-Seq to explore the early transcriptomic response after intoxication with deltamethrin in a population of T. infestans which presents low resistance to pyrethroids. We were able to assemble a complete transcriptome of this vector and found evidence of differentially expressed genes belonging to diverse families such as chemosensory and odorant-binding proteins, ABC transporters and heat-shock proteins. Moreover, genes related to transcription and translation, energetic metabolism and cuticle rearrangements were also modulated. Finally, we characterized the repertoire of previously uncharacterized detoxification-related gene families in T. infestans and Rhodnius prolixus. CONCLUSIONS AND SIGNIFICANCE: Our work contributes to the understanding of the detoxification response in vectors of Chagas disease. Given the absence of an annotated genome from T. infestans, the analysis presented here constitutes a resource for molecular and physiological studies in this species. The results increase the knowledge on detoxification processes in vectors of Chagas disease, and provide relevant information to explore undescribed potential insecticide resistance mechanisms in populations of these insects.


Assuntos
Doença de Chagas , Inseticidas , Piretrinas , Triatoma , Animais , Resistência a Inseticidas/genética , Inseticidas/metabolismo , Inseticidas/farmacologia , Nitrilas/metabolismo , Piretrinas/metabolismo , Piretrinas/farmacologia , Transcriptoma
6.
iScience ; 25(7): 104502, 2022 Jul 15.
Artigo em Inglês | MEDLINE | ID: mdl-35720264

RESUMO

Salts are essential nutrients required for many physiological processes, and accordingly, their composition and concentration are tightly regulated. Taste is the ultimate sensory modality involved in resource quality assessment, resulting in acceptance or rejection. Here we found that high salt concentrations elicit feeding avoidance in the blood-sucking bug Rhodnius prolixus and elucidate the molecular and neurophysiological mechanisms involved. We found that high-salt avoidance is mediated by a salt-sensitive antennal gustatory receptor neuron (GRN). Using RNAi, we demonstrate that this process requires two amiloride-sensitive pickpocket channels (PPKs; Rpro PPK014276 and Rpro PPK28) expressed within these cells. We found that antennal GRNs project to the insect primary olfactory center, the antennal lobes, revealing these centers as potential sites for the integration of taste and olfactory host-derived cues. Moreover, the identification of the gustatory basis of high-salt detection in a hematophagous insect suggests novel targets for the prevention of biting and feeding.

7.
Insect Biochem Mol Biol ; 140: 103704, 2022 01.
Artigo em Inglês | MEDLINE | ID: mdl-34942331

RESUMO

Triatomine bugs are the blood feeding insect vectors transmitting Chagas disease to humans, a neglected tropical disease that affects over 8 million people, mainly in Latin America. The behavioral responses to host cues and bug signals in Rhodnius prolixus are state dependent, i.e., they vary as a function of post-ecdysis age. At the molecular level, these changes in behavior are probably due to a modulation of peripheral and central processes. In the present study, we report a significant modulation of the expression of a large set of sensory-related genes. Results were generated by means of antennal transcriptomes of 5th instar larvae along the first week (days 0, 2, 4, 6 and 8) after ecdysis sequenced using the Illumina HiSeq platform. Significant age-induced changes in transcript abundance were established for more than 6120 genes (54,7% of 11,186 genes expressed) in the antenna of R. prolixus. This was especially true between the first two days after ecdysis when more than 2500 genes had their expression significantly altered. In contrast, expression profiles were almost identical between day 6 and 8, with only a few genes showing significant modulation of their expression. A total of 86 sensory receptors, odorant carriers and odorant degrading enzymes were significantly modulated across age points and clustered into three distinct expression profiles. The set of sensory genes whose expression increased with age (profile 3) may include candidates underlying the increased responsiveness to host cues shown by R. prolixus during the first days after molting. For the first time, we describe the maturation process undergone at the molecular level by the peripheral sensory system of a hemimetabolous insect.


Assuntos
Antenas de Artrópodes , Genes de Insetos , Rhodnius , Órgãos dos Sentidos , Animais , Doença de Chagas/transmissão , Perfilação da Expressão Gênica , Insetos Vetores/genética , Insetos Vetores/metabolismo , Larva/genética , Larva/metabolismo , Odorantes , Receptores Odorantes/genética , Receptores Odorantes/metabolismo , Rhodnius/genética , Rhodnius/metabolismo , Órgãos dos Sentidos/embriologia , Órgãos dos Sentidos/fisiologia , Olfato/genética , Transcriptoma
8.
Genome Biol Evol ; 13(9)2021 09 01.
Artigo em Inglês | MEDLINE | ID: mdl-34390578

RESUMO

Insect pickpocket (PPK) receptors mediate diverse functions, among them the detection of mechano- and chemo-sensory stimuli. Notwithstanding their relevance, studies on their evolution only focused on Drosophila. We have analyzed the genomes of 26 species of eight orders including holometabolous and hemimetabolous insects (Blattodea, Orthoptera, Hemiptera, Phthiraptera, Hymenoptera, Lepidoptera, Coleoptera, and Diptera), to characterize the evolution of this gene family. PPKs were detected in all genomes analyzed, with 578 genes distributed in seven subfamilies. According to our phylogeny, ppk17 is the most divergent member, composing the new subfamily VII. PPKs evolved under a gene birth-and-death model that generated lineage-specific expansions usually located in clusters, while purifying selection affected several orthogroups. Subfamily V was the largest, including a mosquito-specific expansion that can be considered a new target for pest control. PPKs present a high gene turnover generating considerable variation. On one hand, Musca domestica (59), Aedes albopictus (51), Culex quinquefasciatus (48), and Blattella germanica (41) presented the largest PPK repertoires. On the other hand, Pediculus humanus (only ppk17), bees, and ants (6-9) had the smallest PPK sets. A subset of prevalent PPKs was identified, indicating very conserved functions for these receptors. Finally, at least 20% of the sequences presented calmodulin-binding motifs, suggesting that these PPKs may amplify sensory responses similarly as proposed for Drosophila melanogaster ppk25. Overall, this work characterized the evolutionary history of these receptors revealing relevant unknown gene sequence features and clade-specific expansions.


Assuntos
Drosophila melanogaster , Evolução Molecular , Animais , Abelhas/genética , Drosophila melanogaster/genética , Genes de Insetos , Insetos/genética , Filogenia
9.
PLoS Negl Trop Dis ; 15(7): e0009587, 2021 07.
Artigo em Inglês | MEDLINE | ID: mdl-34270558

RESUMO

BACKGROUND: Aedes aegypti (L.) is an urban mosquito, vector of several arboviruses that cause severe diseases in hundreds of million people each year. The resistance to synthetic insecticides developed by Ae. aegypti populations worldwide has contributed to failures in vector control campaigns, increasing the impact of arbovirus diseases. In this context, plant-derived essential oils with larvicidal activity could be an attractive alternative for vector control. However, the mode of action and the detoxificant response of mosquitoes to plant derived compounds have not been established, impairing the optimization of their use. METHODS AND FINDINGS: Here we compare gene expression in Ae. aegypti larvae after 14 hrs of exposure to Eucalyptus camaldulensis essential oil with a control group exposed to vehicle (acetone) for the same lapse, by using RNA-Seq. We found differentially expressed genes encoding for cuticle proteins, fatty-acid synthesis, membrane transporters and detoxificant related gene families (i.e. heat shock proteins, cytochromes P450, glutathione transferases, UDP-glycosyltransferases and ABC transporters). Finally, our RNA-Seq and molecular docking results provide evidence pointing to a central involvement of chemosensory proteins in the detoxificant response in mosquitoes. CONCLUSIONS AND SIGNIFICANCE: Our work contributes to the understanding of the physiological response of Ae. aegypti larvae to an intoxication with a natural toxic distilled from Eucalyptus leafs. The results suggest an involvement of most of the gene families associated to detoxification of xenobiotics in insects. Noteworthy, this work provides important information regarding the implication of chemosensory proteins in the detoxification of a natural larvicide. Understanding the mode of detoxification of Eucalyptus distilled compounds could contribute to their implementation as a tool in mosquito control.


Assuntos
Aedes/efeitos dos fármacos , Eucalyptus/química , Simulação de Acoplamento Molecular , Óleos Voláteis/farmacologia , Óleos de Plantas/farmacologia , Transcriptoma , Aedes/metabolismo , Animais , Sequência de Bases , Biologia Computacional , Sistema Enzimático do Citocromo P-450/genética , Sistema Enzimático do Citocromo P-450/metabolismo , Regulação da Expressão Gênica/efeitos dos fármacos , Proteínas de Choque Térmico/genética , Proteínas de Choque Térmico/metabolismo , Proteínas de Insetos/genética , Proteínas de Insetos/metabolismo , Larva/efeitos dos fármacos , Modelos Moleculares , Óleos Voláteis/química , Óleos de Plantas/química , Conformação Proteica , RNA/genética
10.
BMC Genomics ; 21(1): 101, 2020 Jan 30.
Artigo em Inglês | MEDLINE | ID: mdl-32000664

RESUMO

BACKGROUND: Rhodnius prolixus has become a model for revealing the molecular bases of insect sensory biology due to the publication of its genome and its well-characterized behavioural repertoire. Gene expression modulation underlies behaviour-triggering processes at peripheral and central levels. Still, the regulation of sensory-related gene transcription in sensory organs is poorly understood. Here we study the genetic bases of plasticity in antennal sensory function, using R. prolixus as an insect model. RESULTS: Antennal expression of neuromodulatory genes such as those coding for neuropeptides, neurohormones and their receptors was characterized in fifth instar larvae and female and male adults by means of RNA-Sequencing (RNA-Seq). New nuclear receptor and takeout gene sequences were identified for this species, as well as those of enzymes involved in the biosynthesis and processing of neuropeptides and biogenic amines. CONCLUSIONS: We report a broad repertoire of neuromodulatory and neuroendocrine-related genes expressed in the antennae of R. prolixus and suggest that they may serve as the local basis for modulation of sensory neuron physiology. Diverse neuropeptide precursor genes showed consistent expression in the antennae of all stages studied. Future studies should characterize the role of these modulatory components acting over antennal sensory processes to assess the relative contribution of peripheral and central regulatory systems on the plastic expression of insect behaviour.


Assuntos
Perfilação da Expressão Gênica/veterinária , Proteínas de Insetos/genética , Rhodnius/crescimento & desenvolvimento , Animais , Antenas de Artrópodes/química , Feminino , Regulação da Expressão Gênica no Desenvolvimento , Larva , Masculino , Neuropeptídeos/genética , Neurotransmissores/genética , Filogenia , Receptores de Neuropeptídeos/genética , Receptores de Neurotransmissores/genética , Rhodnius/genética , Análise de Sequência de RNA/veterinária
11.
Curr Opin Insect Sci ; 34: 80-84, 2019 08.
Artigo em Inglês | MEDLINE | ID: mdl-31247423

RESUMO

Sensory processes represent an information gathering interface between animals and their surrounding world. Therefore, they serve to scan the environment for resources and threats. The behavior of kissing bugs has been studied to aid their control because they transmit Chagas disease to humans. Besides, a few triatomines represent important insect models since Wigglesworth times. These hematophagous insects rely on different sensory systems to scan their environment for blood-sources, mating partners, and hiding places. The study of the molecular bases of sensory processes has undergone a dramatic progress due the advent of new technologies allowing mass-sequencing of genes. Here, we focus on reviewing the fundamental knowledge gathered to date about the molecular bases of kissing bug sensory processes.


Assuntos
Insetos Vetores/fisiologia , Rhodnius/fisiologia , Sensação , Células Receptoras Sensoriais/fisiologia , Animais
12.
PLoS Negl Trop Dis ; 11(2): e0005313, 2017 02.
Artigo em Inglês | MEDLINE | ID: mdl-28199333

RESUMO

BACKGROUND: Triatomine insects are vectors of Trypanosoma cruzi, a protozoan parasite that is the causative agent of Chagas' disease. This is a neglected disease affecting approximately 8 million people in Latin America. The existence of diverse pyrethroid resistant populations of at least two species demonstrates the potential of triatomines to develop high levels of insecticide resistance. Therefore, the incorporation of strategies for resistance management is a main concern for vector control programs. Three enzymatic superfamilies are thought to mediate xenobiotic detoxification and resistance: Glutathione Transferases (GSTs), Cytochromes P450 (CYPs) and Carboxyl/Cholinesterases (CCEs). Improving our knowledge of key triatomine detoxification enzymes will strengthen our understanding of insecticide resistance processes in vectors of Chagas' disease. METHODS AND FINDINGS: The discovery and description of detoxification gene superfamilies in normalized transcriptomes of three triatomine species: Triatoma dimidiata, Triatoma infestans and Triatoma pallidipennis is presented. Furthermore, a comparative analysis of these superfamilies among the triatomine transcriptomes and the genome of Rhodnius prolixus, also a triatomine vector of Chagas' disease, and other well-studied insect genomes was performed. The expression pattern of detoxification genes in R. prolixus transcriptomes from key organs was analyzed. The comparisons reveal gene expansions in Sigma class GSTs, CYP3 in CYP superfamily and clade E in CCE superfamily. Moreover, several CYP families identified in these triatomines have not yet been described in other insects. Conversely, several groups of insecticide resistance related enzymes within each enzyme superfamily are reduced or lacking in triatomines. Furthermore, our qRT-PCR results showed an increase in the expression of a CYP4 gene in a T. infestans population resistant to pyrethroids. These results could point to an involvement of metabolic detoxification mechanisms on the high levels of pyrethroid resistance detected in triatomines from the Gran Chaco ecoregion. CONCLUSIONS AND SIGNIFICANCE: Our results help to elucidate the potential insecticide resistance mechanisms in vectors of Chagas' disease and provide new relevant information for this field. This study shows that metabolic resistance might be a contributing cause of the high pyrethroid resistance observed in wild T. infestans populations from the Gran Chaco ecoregion, area in which although subjected to intense pyrethroid treatments, vector control has failed. This study opens new avenues for further functional studies on triatomine detoxification mechanisms.


Assuntos
Genoma de Inseto , Proteínas de Insetos/genética , Insetos Vetores/efeitos dos fármacos , Insetos Vetores/genética , Inseticidas/farmacologia , Triatoma/efeitos dos fármacos , Triatoma/genética , Animais , Sistema Enzimático do Citocromo P-450/genética , Sistema Enzimático do Citocromo P-450/metabolismo , Genômica , Proteínas de Insetos/química , Proteínas de Insetos/metabolismo , Insetos Vetores/classificação , Insetos Vetores/metabolismo , Filogenia , Triatoma/classificação , Triatoma/metabolismo
13.
Sci Rep ; 7: 40049, 2017 01 06.
Artigo em Inglês | MEDLINE | ID: mdl-28059141

RESUMO

The triatomine bug Rhodnius prolixus is a main vector of Chagas disease, which affects several million people, mostly in Latin-America. Host searching, pheromone communication, and microclimatic preferences are aspects of its behaviour that depend on multimodal sensory inputs. The molecular bases of these sensory processes are largely unknown. The expression levels of genes transcribed in antennae were compared between 5th instar larvae, and female and male adults by means of RNA-Seq. The antennae of R. prolixus showed increased expression of several chemosensory-related genes in imaginal bugs, while both sexes had similar expression patterns for most target genes. Few cases suggest involvement of target genes in sexually dimorphic functions. Most odorant and ionotropic receptor genes seemed to be expressed in all libraries. OBPs and CSPs showed very high expression levels. Other sensory-related genes such as TRPs, PPKs and mechanoreceptors had consistent levels of expression in all libraries. Our study characterises most of the sensory gene repertoire of these insects, opening an avenue for functional genetics studies. The increase in expression of chemosensory genes suggests an enhanced role in adult bugs. This knowledge allows developing new behaviour interfering strategies, increasing the options for translational research in the vector control field.


Assuntos
Antenas de Artrópodes/fisiologia , Insetos Vetores/fisiologia , Receptores Odorantes/genética , Rhodnius/fisiologia , Animais , Feminino , Perfilação da Expressão Gênica , Insetos Vetores/genética , Larva/genética , Larva/fisiologia , Masculino , Feromônios/metabolismo , Rhodnius/genética , Análise de Sequência de RNA
14.
Insect Biochem Mol Biol ; 69: 71-81, 2016 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-26003917

RESUMO

Rhodnius prolixus is a triatomine bug acting as a relevant vector of Chagas disease for which the genome sequence has been recently made available. Based on this information, a set of olfactory (ORs) and ionotropic receptor (IRs) genes potentially related to olfactory processes was characterized, and the expression patterns along bug development and in different structures potentially involved in promoting chemosensory-mediated behaviors were studied. For this, diverse bioinformatic procedures were used to validate gene models analyzing their structural and functional features and designing specific primers. Evolutionary relationships among R. prolixus olfactory coreceptors (RproOrco, RproIR25a, RproIR8a and RproIR76b) and their orthologues from other insects were shown to have mostly good bootstrap support values in phylogenetic trees. Moreover, antennal expression was confirmed for most genes included in the study. Both ORs and IRs showed antennal expression along the whole development of bugs of this species, with few exceptional receptors showing gradually increasing expression or expression restricted to the antennae of adult bugs. Finally, the expression of most of the selected genes was confirmed in other structures, such as rostri, tarsi, tibial pads and genitalia, which are potentially involved in promoting chemosensory-mediated behaviors. These results are discussed in terms of their relevance to advance in the understanding of the molecular bases of triatomine behavior.


Assuntos
Receptores Odorantes/genética , Rhodnius/genética , Animais , Antenas de Artrópodes/metabolismo , Doença de Chagas/transmissão , Feminino , Perfilação da Expressão Gênica , Insetos Vetores , Masculino , Filogenia , Receptores Ionotrópicos de Glutamato/genética , Rhodnius/embriologia , Rhodnius/crescimento & desenvolvimento , Células Receptoras Sensoriais/metabolismo
15.
PLoS Negl Trop Dis ; 9(8): e0003973, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-26291723

RESUMO

BACKGROUND: As a result of evolution, the biology of triatomines must have been significantly adapted to accommodate trypanosome infection in a complex network of vector-vertebrate-parasite interactions. Arthropod-borne parasites have probably developed mechanisms, largely still unknown, to exploit the vector-vertebrate host interactions to ensure their transmission to suitable hosts. Triatomines exhibit a strong negative phototaxis and nocturnal activity, believed to be important for insect survival against its predators. METHODOLOGY/PRINCIPAL FINDINGS: In this study we quantified phototaxis and locomotion in starved fifth instar nymphs of Rhodnius prolixus infected with Trypanosoma cruzi or Trypanosoma rangeli. T. cruzi infection did not alter insect phototaxis, but induced an overall 20% decrease in the number of bug locomotory events. Furthermore, the significant differences induced by this parasite were concentrated at the beginning of the scotophase. Conversely, T. rangeli modified both behaviors, as it significantly decreased bug negative phototaxis, while it induced a 23% increase in the number of locomotory events in infected bugs. In this case, the significant effects were observed during the photophase. We also investigated the expression of Rpfor, the triatomine ortholog of the foraging gene known to modulate locomotion in other insects, and found a 4.8 fold increase for T. rangeli infected insects. CONCLUSIONS/SIGNIFICANCE: We demonstrated for the first time that trypanosome infection modulates the locomotory activity of the invertebrate host. T. rangeli infection seems to be more broadly effective, as besides affecting the intensity of locomotion this parasite also diminished negative phototaxis and the expression of a behavior-associated gene in the triatomine vector.


Assuntos
Comportamento Animal , Regulação da Expressão Gênica , Insetos Vetores/parasitologia , Atividade Motora , Rhodnius/parasitologia , Trypanosoma/fisiologia , Sequência de Aminoácidos , Animais , Interações Hospedeiro-Parasita , Proteínas de Insetos/genética , Proteínas de Insetos/metabolismo , Insetos Vetores/genética , Insetos Vetores/fisiologia , Luz , Dados de Sequência Molecular , Atividade Motora/genética , Rhodnius/genética , Rhodnius/fisiologia , Trypanosoma cruzi/fisiologia , Trypanosoma rangeli/fisiologia
16.
Parasit Vectors ; 8: 243, 2015 Apr 22.
Artigo em Inglês | MEDLINE | ID: mdl-25896676

RESUMO

BACKGROUND: Quantitative reverse transcription PCR (qRT-PCR) is a robust and accessible method to assay gene expression and to infer gene regulation. Being a chain of procedures, this technique is subject to systematic error due to biological and technical limitations mainly set by the starting material and downstream procedures. Thus, rigorous data normalization is critical to grant reliability and repeatability of gene expression quantification by qRT-PCR. A number of 'housekeeping genes', involved in basic cellular functions, have been commonly used as internal controls for this normalization process. However, these genes could themselves be regulated and must therefore be tested a priori. METHODS: We evaluated eight potential reference genes for their stability as internal controls for RT-qPCR studies of olfactory gene expression in the antennae of Rhodnius prolixus, a Chagas disease vector. The set of genes included were: α-tubulin; ß-actin; Glyceraldehyde-3-phosphate dehydrogenase; Eukaryotic initiation factor 1A; Glutathione-S-transferase; Serine protease; Succinate dehydrogenase; and Glucose-6-phosphate dehydrogenase. Five experimental conditions, including changes in age,developmental stage and feeding status were tested in both sexes. RESULTS: We show that the evaluation of candidate reference genes is necessary for each combination of sex, tissue and physiological condition analyzed in order to avoid inconsistent results and conclusions. Although, Normfinder and geNorm software yielded different results between males and females, five genes (SDH, Tub, GAPDH, Act and G6PDH) appeared in the first positions in all rankings obtained. By using gene expression data of a single olfactory coreceptor gene as an example, we demonstrated the extent of changes expected using different internal standards. CONCLUSIONS: This work underlines the need for a rigorous selection of internal standards to grant the reliability of normalization processes in qRT-PCR studies. Furthermore, we show that particular physiological or developmental conditions require independent evaluation of a diverse set of potential reference genes.


Assuntos
Entomologia/métodos , Perfilação da Expressão Gênica/métodos , Genes Essenciais , Reação em Cadeia da Polimerase em Tempo Real/métodos , Padrões de Referência , Rhodnius/genética , Olfato/genética , Animais , Antenas de Artrópodes/fisiologia , Rhodnius/fisiologia
17.
Mem Inst Oswaldo Cruz ; 108 Suppl 1: 63-73, 2013.
Artigo em Inglês | MEDLINE | ID: mdl-24473804

RESUMO

Triatomines have been important model organisms for behavioural research. Diverse reports about triatomine host search, pheromone communication in the sexual, shelter and alarm contexts, daily cycles of activity, refuge choice and behavioural plasticity have been published in the last two decades. In recent times, a variety of molecular genetics techniques has allowed researchers to investigate elaborate and complex questions about the genetic bases of the physiology of insects. This, together with the current characterisation of the genome sequence of Rhodnius prolixus allows the resurgence of this excellent insect physiology model in the omics era. In the present revision, we suggest that studying the molecular basis of behaviour and sensory ecology in triatomines will promote a deeper understanding of fundamental aspects of insect and, particularly, vector biology. This will allow uncovering unknown features of essential insect physiology questions for a hemimetabolous model organism, promoting more robust comparative studies of insect sensory function and cognition.


Assuntos
Comportamento Animal/fisiologia , Genoma de Inseto/genética , Insetos Vetores/genética , Triatominae/genética , Animais , Relógios Circadianos/genética , Ritmo Circadiano/genética , Locomoção , Feromônios/genética , Rhodnius/genética , Navegação Espacial
18.
Mem. Inst. Oswaldo Cruz ; 108(supl.1): 63-73, 2013. tab, graf
Artigo em Inglês | LILACS | ID: lil-697828

RESUMO

Triatomines have been important model organisms for behavioural research. Diverse reports about triatomine host search, pheromone communication in the sexual, shelter and alarm contexts, daily cycles of activity, refuge choice and behavioural plasticity have been published in the last two decades. In recent times, a variety of molecular genetics techniques has allowed researchers to investigate elaborate and complex questions about the genetic bases of the physiology of insects. This, together with the current characterisation of the genome sequence of Rhodnius prolixus allows the resurgence of this excellent insect physiology model in the omics era. In the present revision, we suggest that studying the molecular basis of behaviour and sensory ecology in triatomines will promote a deeper understanding of fundamental aspects of insect and, particularly, vector biology. This will allow uncovering unknown features of essential insect physiology questions for a hemimetabolous model organism, promoting more robust comparative studies of insect sensory function and cognition.


Assuntos
Animais , Comportamento Animal/fisiologia , Genoma de Inseto/genética , Insetos Vetores/genética , Triatominae/genética , Relógios Circadianos/genética , Ritmo Circadiano/genética , Locomoção , Feromônios/genética , Rhodnius/genética , Navegação Espacial
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