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1.
Arch Insect Biochem Physiol ; 115(4): e22106, 2024 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-38597092

RESUMEN

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.


Asunto(s)
Neuropéptidos , Rhodnius , Triatoma , Animales , Rhodnius/genética , Rhodnius/metabolismo , Neuropéptidos/genética , Neuropéptidos/metabolismo , Transcriptoma , Muda
2.
PLoS One ; 18(3): e0282490, 2023.
Artículo en Inglés | MEDLINE | ID: mdl-36867641

RESUMEN

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.


Asunto(s)
Enfermedad de Chagas , Rhodnius , Animales , Encéfalo , Conocimiento , Ninfa , Expresión Génica
3.
BMC Genomics ; 23(1): 861, 2022 Dec 30.
Artículo en Inglés | MEDLINE | ID: mdl-36585617

RESUMEN

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.


Asunto(s)
Enfermedad de Chagas , Triatoma , Animales , Humanos , Triatoma/genética , Triatoma/metabolismo , Transcriptoma , Bolivia , Resistencia a los Insecticidas
4.
Parasitology ; 149(2): 155-160, 2022 02.
Artículo en Inglés | MEDLINE | ID: mdl-35234603

RESUMEN

Trypanosoma rangeli is a protozoan that infects triatomines and mammals in Latin America, sharing hosts with Trypanosoma cruzi, the etiological agent of Chagas disease. Trypanosoma rangeli does not cause disease to humans but is strongly pathogenic to its invertebrate hosts, increasing mortality rates and affecting bug development and reproductive success. We have previously shown that this parasite is also capable of inducing a general increase in the locomotory activity of its vector Rhodnius prolixus in the absence of host cues. In this work, we have evaluated whether infection impacts the insect­vertebrate host interaction. For this, T. rangeli-infected and uninfected R. prolixus nymphs were released in glass arenas offering single shelters. After a 3-day acclimatization, a caged mouse was introduced in each arena and shelter use and predation rates were evaluated. Trypanosoma rangeli infection affected all parameters analysed. A larger number of infected bugs was found outside shelters, both in the absence and presence of a host. Infected bugs also endured greater predation rates, probably because of an increased number of individuals that attempted to feed. Interestingly, mice that predated on infected bugs did not develop T. rangeli infection, suggesting that the oral route is not effective for these parasites, at least in our system. Finally, a smaller number of infected bugs succeeded in feeding in this context. We suggest that, although T. rangeli is not transmitted orally, an increase in the proportion of foraging individuals would promote greater parasite transmission rates through an increased frequency of very effective infected-bug bites.


Asunto(s)
Rhodnius , Trypanosoma cruzi , Trypanosoma rangeli , Trypanosoma , Animales , Insectos Vectores/parasitología , Mamíferos , Ratones , Conducta Predatoria , Rhodnius/parasitología
5.
Insect Biochem Mol Biol ; 140: 103704, 2022 01.
Artículo en Inglés | MEDLINE | ID: mdl-34942331

RESUMEN

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.


Asunto(s)
Antenas de Artrópodos , Genes de Insecto , Rhodnius , Órganos de los Sentidos , Animales , Enfermedad de Chagas/transmisión , Perfilación de la Expresión Génica , Insectos Vectores/genética , Insectos Vectores/metabolismo , Larva/genética , Larva/metabolismo , Odorantes , Receptores Odorantes/genética , Receptores Odorantes/metabolismo , Rhodnius/genética , Rhodnius/metabolismo , Órganos de los Sentidos/embriología , Órganos de los Sentidos/fisiología , Olfato/genética , Transcriptoma
6.
Genome Biol Evol ; 13(9)2021 09 01.
Artículo en Inglés | MEDLINE | ID: mdl-34390578

RESUMEN

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.


Asunto(s)
Drosophila melanogaster , Evolución Molecular , Animales , Abejas/genética , Drosophila melanogaster/genética , Genes de Insecto , Insectos/genética , Filogenia
7.
PLoS Negl Trop Dis ; 15(7): e0009570, 2021 07.
Artículo en Inglés | MEDLINE | ID: mdl-34197458

RESUMEN

Triatomine bugs aggregate with conspecifics inside shelters during daylight hours. At dusk, they leave their refuges searching for hosts on which to blood feed. After finding a host, triatomines face the threat of being killed, because hosts often prey on them. As it is known that many parasites induce the predation of intermediate hosts to promote transmission, and that ingestion of Trypanosoma cruzi-infected bugs represents a very effective means for mammal infection, we hypothesized that trypanosomes induce infected bugs to take increased risk, and, as a consequence, be predated when approaching a host. Therefore, we evaluated whether the predation risk and predation rates endured by Rhodnius prolixus increase when infected with T. cruzi. Assays were performed in square glass arenas offering one central refuge to infected and uninfected 5th instar nymphs. A caged mouse was introduced in each arena after a three-day acclimation interval to activate sheltered insects and induce them to approach it. As hypothesized, a significantly higher proportion of infected insects was predated when compared with uninfected ones (36% and 19%, respectively). Indeed, T. cruzi-infected bugs took higher risk (Approximation Index = 0.642) when compared with healthy ones (Approximation Index = 0.302) and remained outside the shelters when the host was removed from the arena. Our results show that infection by T. cruzi induces bugs to assume higher risk and endure higher predation rates. We reveal a hitherto unknown trypanosome-vector interaction process that increases infected bug predation, promoting increased rates of robust oral transmission. The significant consequences of the mechanism revealed here make it a fundamental component for the resilient maintenance of sylvatic, peridomestic and domestic cycles.


Asunto(s)
Rhodnius/parasitología , Trypanosoma cruzi/fisiología , Animales , Pollos , Especificidad del Huésped , Interacciones Huésped-Parásitos , Ratones , Ninfa/parasitología , Conducta Predatoria
8.
J Insect Physiol ; 122: 104021, 2020 04.
Artículo en Inglés | MEDLINE | ID: mdl-32035953

RESUMEN

The triatomine bug Rhodnius prolixus is a main vector of Chagas disease, which affects several million people in Latin-America. These nocturnal insects spend most of their locomotory activity during the first hours of the scotophase searching for suitable hosts. In this study we used multivariate analysis to characterize spontaneous locomotory activity profiles presented by 5th instar nymphs. In addition, we investigated whether sex and the expression of the foraging (Rpfor) gene could modulate this behavioral trait. Hierarchical Clustering and Redundancy Analyses detected individuals with distinct locomotory profiles. In addition to a great variation in locomotory intensity, we found that a proportion of nymphs walked during unusual time intervals. Locomotory activity profiles were mostly affected by the cumulative activity expressed by the nymphs. These effects promoted by cumulative activity were in turn influenced by nymph sex. Sex and the Rpfor expression had a significant influence on the profiles, as well as in the levels of total activity. In conclusion, the locomotory profiles evinced by the multivariate analyses suggest the co-existence of different foraging strategies in bugs. Additionally, we report sex-specific effects on the locomotion patterns of 5th instar R. prolixus, which are apparently modulated by the differential expression of the Rpfor gene.


Asunto(s)
Locomoción , Rhodnius/fisiología , Animales , Enfermedad de Chagas/transmisión , Conducta Alimentaria , Genes de Insecto , Insectos Vectores/genética , Insectos Vectores/fisiología , Locomoción/genética , Locomoción/fisiología , Ninfa/genética , Ninfa/fisiología , Rhodnius/genética , Factores Sexuales , Transcriptoma
9.
BMC Genomics ; 21(1): 101, 2020 Jan 30.
Artículo en Inglés | MEDLINE | ID: mdl-32000664

RESUMEN

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.


Asunto(s)
Perfilación de la Expresión Génica/veterinaria , Proteínas de Insectos/genética , Rhodnius/crecimiento & desarrollo , Animales , Antenas de Artrópodos/química , Femenino , Regulación del Desarrollo de la Expresión Génica , Larva , Masculino , Neuropéptidos/genética , Neurotransmisores/genética , Filogenia , Receptores de Neuropéptido/genética , Receptores de Neurotransmisores/genética , Rhodnius/genética , Análisis de Secuencia de ARN/veterinaria
10.
Sci Rep ; 7: 40049, 2017 01 06.
Artículo en Inglés | MEDLINE | ID: mdl-28059141

RESUMEN

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.


Asunto(s)
Antenas de Artrópodos/fisiología , Insectos Vectores/fisiología , Receptores Odorantes/genética , Rhodnius/fisiología , Animales , Femenino , Perfilación de la Expresión Génica , Insectos Vectores/genética , Larva/genética , Larva/fisiología , Masculino , Feromonas/metabolismo , Rhodnius/genética , Análisis de Secuencia de ARN
11.
J Insect Physiol ; 97: 66-76, 2017.
Artículo en Inglés | MEDLINE | ID: mdl-27401496

RESUMEN

Triatomines are hematophagous insects that feed on the blood of vertebrates from different taxa, but can occasionally also take fluids from invertebrate hosts, including other insects. During the blood ingestion process, these insects can acquire diverse parasites that can later be transmitted to susceptible vertebrates if they complete their development inside bugs. Trypanosoma cruzi, the etiological agent of Chagas disease, and Trypanosoma rangeli are protozoan parasites transmitted by triatomines, the latter only transmitted by Rhodnius spp. The present work makes an extensive revision of studies evaluating triatomine-trypanosome interaction, with special focus on Rhodnius prolixus interacting with the two parasites. The sequences of events encompassing the development of these trypanosomes inside bugs and the consequent responses of insects to this infection, as well as many pathological effects produced by the parasites are discussed.


Asunto(s)
Rhodnius/fisiología , Rhodnius/parasitología , Trypanosoma cruzi/crecimiento & desarrollo , Trypanosoma rangeli/crecimiento & desarrollo , Animales , Trypanosoma cruzi/fisiología , Trypanosoma rangeli/fisiología
12.
Parasitology ; 143(4): 434-43, 2016 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-26818093

RESUMEN

Trypanosoma cruzi, the etiological agent of Chagas disease, is ingested by triatomines during their bloodmeal on an infected mammal. Aiming to investigate the development and differentiation of T. cruzi inside the intestinal tract of Rhodnius prolixus at the beginning of infection we fed insects with cultured epimastigotes and blood trypomastigotes from infected mice to determine the amount of recovered parasites after ingestion. Approximately 20% of the ingested parasites was found in the insect anterior midgut (AM) 3 h after feeding. Interestingly, a significant reduction (80%) in the numbers of trypomastigotes was observed after 24 h of infection suggesting that parasites were killed in the AM. Moreover, few parasites were found in that intestinal portion after 96 h of infection. The evaluation of the numbers of parasites in the posterior midgut (PM) at the same periods showed a reduced parasite load, indicating that parasites were not moving from the AM. Additionally, incubation of blood trypomastigotes with extracts from R. prolixus AMs revealed that components of this tissue could induce significant death of T. cruzi. Finally, we observed that differentiation from trypomastigotes to epimastigotes is not completed in the AM; instead we suggest that trypomastigotes change to intermediary forms before their migration to the PM, where differentiation to epimastigotes takes place. The present work clarifies controversial points concerning T. cruzi development in insect vector, showing that parasite suffers a drastic decrease in population size before epimastigonesis accomplishment in PM.


Asunto(s)
Enfermedad de Chagas/parasitología , Insectos Vectores/parasitología , Rhodnius/parasitología , Trypanosoma cruzi/crecimiento & desarrollo , Análisis de Varianza , Animales , Enfermedad de Chagas/sangre , Enfermedad de Chagas/transmisión , ADN Protozoario/química , ADN Protozoario/aislamiento & purificación , Ratones , Ninfa/parasitología , Reacción en Cadena en Tiempo Real de la Polimerasa , Trypanosoma cruzi/genética
13.
Insect Biochem Mol Biol ; 69: 71-81, 2016 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-26003917

RESUMEN

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.


Asunto(s)
Receptores Odorantes/genética , Rhodnius/genética , Animales , Antenas de Artrópodos/metabolismo , Enfermedad de Chagas/transmisión , Femenino , Perfilación de la Expresión Génica , Insectos Vectores , Masculino , Filogenia , Receptores Ionotrópicos de Glutamato/genética , Rhodnius/embriología , Rhodnius/crecimiento & desarrollo , Células Receptoras Sensoriales/metabolismo
14.
PLoS Negl Trop Dis ; 9(8): e0003973, 2015.
Artículo en Inglés | MEDLINE | ID: mdl-26291723

RESUMEN

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.


Asunto(s)
Conducta Animal , Regulación de la Expresión Génica , Insectos Vectores/parasitología , Actividad Motora , Rhodnius/parasitología , Trypanosoma/fisiología , Secuencia de Aminoácidos , Animales , Interacciones Huésped-Parásitos , Proteínas de Insectos/genética , Proteínas de Insectos/metabolismo , Insectos Vectores/genética , Insectos Vectores/fisiología , Luz , Datos de Secuencia Molecular , Actividad Motora/genética , Rhodnius/genética , Rhodnius/fisiología , Trypanosoma cruzi/fisiología , Trypanosoma rangeli/fisiología
15.
Parasit Vectors ; 8: 243, 2015 Apr 22.
Artículo en Inglés | MEDLINE | ID: mdl-25896676

RESUMEN

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.


Asunto(s)
Entomología/métodos , Perfilación de la Expresión Génica/métodos , Genes Esenciales , Reacción en Cadena en Tiempo Real de la Polimerasa/métodos , Estándares de Referencia , Rhodnius/genética , Olfato/genética , Animales , Antenas de Artrópodos/fisiología , Rhodnius/fisiología
16.
PLoS One ; 9(8): e105255, 2014.
Artículo en Inglés | MEDLINE | ID: mdl-25136800

RESUMEN

The insect Rhodnius prolixus is responsible for the transmission of Trypanosoma cruzi, which is the etiological agent of Chagas disease in areas of Central and South America. Besides this, it can be infected by other trypanosomes such as Trypanosoma rangeli. The effects of these parasites on vectors are poorly understood and are often controversial so here we focussed on possible negative effects of these parasites on the reproductive performance of R. prolixus, specifically comparing infected and uninfected couples. While T. cruzi infection did not delay pre-oviposition time of infected couples at either temperature tested (25 and 30°C) it did, at 25°C, increase the e-value in the second reproductive cycle, as well as hatching rates. Meanwhile, at 30°C, T. cruzi infection decreased the e-value of insects during the first cycle and also the fertility of older insects. When couples were instead infected with T. rangeli, pre-oviposition time was delayed, while reductions in the e-value and hatching rate were observed in the second and third cycles. We conclude that both T. cruzi and T. rangeli can impair reproductive performance of R. prolixus, although for T. cruzi, this is dependent on rearing temperature and insect age. We discuss these reproductive costs in terms of potential consequences on triatomine behavior and survival.


Asunto(s)
Insectos Vectores/parasitología , Rhodnius/parasitología , Trypanosoma cruzi/fisiología , Trypanosoma rangeli/fisiología , Animales , Femenino , Interacciones Huésped-Patógeno , Insectos Vectores/fisiología , Masculino , Oviposición , Reproducción , Rhodnius/fisiología
17.
Mem Inst Oswaldo Cruz ; 108 Suppl 1: 34-47, 2013.
Artículo en Inglés | MEDLINE | ID: mdl-24473801

RESUMEN

Many arthropod species have adopted vertebrate blood as their main food source. Blood is rich in nutrients and, except for the presence of parasites, sterile. However, this food source is not freely available, nor is obtaining it devoid of risk. It circulates inside vessels hidden underneath the skin of mobile hosts that are able to defend themselves and even predate the insects that try to feed on them. Thus, the haematophagous lifestyle is associated with major morphological, physiological and behavioural adaptations that have accumulated throughout the evolutionary history of the various lineages of blood-sucking arthropods. These adaptations have significant consequences for the evolution of parasites as well as for the epidemiology of vector-transmitted diseases. In this review article, we analyse various aspects of the behaviour of triatomine bugs to illustrate how each behavioural trait represents a particular adaptation to their close association with their hosts, which may easily turn into predators. Our aim is to offer to the reader an up-to-date integrative perspective on the behaviour of Chagas disease vectors and to propose new research avenues to encourage both young and experienced colleagues to explore this aspect of triatomine biology.


Asunto(s)
Conducta Animal/fisiología , Enfermedad de Chagas/transmisión , Insectos Vectores/fisiología , Rhodnius/parasitología , Triatoma/parasitología , Adaptación Fisiológica/fisiología , Animales , Relojes Circadianos , Conducta Alimentaria/fisiología , Interacciones Huésped-Parásitos/fisiología , Calor , Odorantes , Trypanosoma cruzi/fisiología
18.
Mem Inst Oswaldo Cruz ; 108 Suppl 1: 63-73, 2013.
Artículo en Inglés | MEDLINE | ID: mdl-24473804

RESUMEN

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.


Asunto(s)
Conducta Animal/fisiología , Genoma de los Insectos/genética , Insectos Vectores/genética , Triatominae/genética , Animales , Relojes Circadianos/genética , Ritmo Circadiano/genética , Locomoción , Feromonas/genética , Rhodnius/genética , Navegación Espacial
19.
Mem. Inst. Oswaldo Cruz ; 108(supl.1): 34-47, 2013. graf
Artículo en Inglés | LILACS | ID: lil-697825

RESUMEN

Many arthropod species have adopted vertebrate blood as their main food source. Blood is rich in nutrients and, except for the presence of parasites, sterile. However, this food source is not freely available, nor is obtaining it devoid of risk. It circulates inside vessels hidden underneath the skin of mobile hosts that are able to defend themselves and even predate the insects that try to feed on them. Thus, the haematophagous lifestyle is associated with major morphological, physiological and behavioural adaptations that have accumulated throughout the evolutionary history of the various lineages of blood-sucking arthropods. These adaptations have significant consequences for the evolution of parasites as well as for the epidemiology of vector-transmitted diseases. In this review article, we analyse various aspects of the behaviour of triatomine bugs to illustrate how each behavioural trait represents a particular adaptation to their close association with their hosts, which may easily turn into predators. Our aim is to offer to the reader an up-to-date integrative perspective on the behaviour of Chagas disease vectors and to propose new research avenues to encourage both young and experienced colleagues to explore this aspect of triatomine biology.


Asunto(s)
Animales , Conducta Animal/fisiología , Enfermedad de Chagas/transmisión , Insectos Vectores/fisiología , Rhodnius/parasitología , Triatoma/parasitología , Adaptación Fisiológica/fisiología , Relojes Circadianos , Conducta Alimentaria/fisiología , Calor , Interacciones Huésped-Parásitos/fisiología , Odorantes , Trypanosoma cruzi/fisiología
20.
Mem. Inst. Oswaldo Cruz ; 108(supl.1): 63-73, 2013. tab, graf
Artículo en Inglés | LILACS | ID: lil-697828

RESUMEN

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.


Asunto(s)
Animales , Conducta Animal/fisiología , Genoma de los Insectos/genética , Insectos Vectores/genética , Triatominae/genética , Relojes Circadianos/genética , Ritmo Circadiano/genética , Locomoción , Feromonas/genética , Rhodnius/genética , Navegación Espacial
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