RESUMEN
Nestmate recognition in ants is regulated through the detection of cuticular hydrocarbons by odorant receptors (ORs) in the antennae. These ORs are crucial for maintaining colony cohesion that allows invasive ant species to dominate colonized environments. In the invasive Argentine ant, Linepithema humile, ORs regulating nestmate recognition are thought to be present in a clade of nine-exon odorant receptors, but the identity of the specific genes remains unknown. We sought to narrow down the list of candidate genes using transcriptomics and phylostratigraphy. Comparative transcriptomic analyses were conducted on the antennae, head, thorax, and legs of Argentine ant workers. We have identified a set of twenty-one nine-exon odorant receptors enriched in the antennae compared to the other tissues, allowing for downstream verification of whether they can detect Argentine ant cuticular hydrocarbons. Further investigation of these ORs could allow us to further understand the mechanisms underlying nestmate recognition and colony cohesion in ants.
Asunto(s)
Hormigas , Filogenia , Receptores Odorantes , Animales , Hormigas/genética , Hormigas/metabolismo , Receptores Odorantes/genética , Receptores Odorantes/metabolismo , Transcriptoma , Perfilación de la Expresión Génica , Antenas de Artrópodos/metabolismo , Proteínas de Insectos/genética , Proteínas de Insectos/metabolismoRESUMEN
Essential oils (EOs) are natural products currently used to control arthropods, and their interaction with insect odorant-binding proteins (OBPs) is fundamental for the discovery of new repellents. This in silico study aimed to predict the potential of EO components to interact with odorant proteins. A total of 684 EO components from PubChem were docked against 23 odorant binding proteins from Protein Data Bank using AutoDock Vina. The ligands and proteins were optimized using Gaussian 09 and Sybyl-X 2.0, respectively. The nature of the protein-ligand interactions was characterized using LigandScout 4.0, and visualization of the binding mode in selected complexes was carried out by Pymol. Additionally, complexes with the best binding energy in molecular docking were subjected to 500 ns molecular dynamics simulations using Gromacs. The best binding affinity values were obtained for the 1DQE-ferutidine (-11 kcal/mol) and 2WCH-kaurene (-11.2 kcal/mol) complexes. Both are natural ligands that dock onto those proteins at the same binding site as DEET, a well-known insect repellent. This study identifies kaurene and ferutidine as possible candidates for natural insect repellents, offering a potential alternative to synthetic chemicals like DEET.
Asunto(s)
Simulación del Acoplamiento Molecular , Aceites Volátiles , Receptores Odorantes , Receptores Odorantes/química , Receptores Odorantes/metabolismo , Aceites Volátiles/química , Animales , Proteínas de Insectos/química , Proteínas de Insectos/metabolismo , Simulación de Dinámica Molecular , Repelentes de Insectos/química , Ligandos , Relación Estructura-Actividad CuantitativaRESUMEN
Reception of chemical information from the environment is crucial for insects' survival and reproduction. The chemosensory reception mainly occurs by the antennae and mouth parts of the insect, when the stimulus contacts the chemoreceptors located within the sensilla. Chemosensory receptor genes have been well-studied in some social hymenopterans such as ants, honeybees, and wasps. However, although stingless bees are the most representative group of eusocial bees, little is known about their odorant, gustatory, and ionotropic receptor genes. Here, we analyze the transcriptome of the proboscis and antennae of the stingless bee Tetragonisca fiebrigi. We identified and annotated 9 gustatory and 15 ionotropic receptors. Regarding the odorant receptors, we identified 204, and we were able to annotate 161 of them. In addition, we compared the chemosensory receptor genes of T. fiebrigi with those annotated for other species of Hymenoptera. We found that T. fiebrigi showed the largest number of odorant receptors compared with other bees. Genetic expansions were identified in the subfamilies 9-exon, which was also expanded in ants and paper wasps; in G02A, including receptors potentially mediating social behavior; and in GUnC, which has been related to pollen and nectar scent detection. Our study provides the first report of chemosensory receptor genes in T. fiebrigi and represents a resource for future molecular and physiological research in this and other stingless bee species.
Asunto(s)
Receptores Odorantes , Animales , Abejas/genética , Abejas/fisiología , Receptores Odorantes/genética , Transcriptoma , Filogenia , Proteínas de Insectos/genética , Proteínas de Insectos/metabolismo , Genes de Insecto , Anotación de Secuencia Molecular , Perfilación de la Expresión GénicaRESUMEN
Chemoreception through odorant receptors (ORs), ionotropic receptors (IRs) and gustatory receptors (GRs) represents the functions of key proteins in the chemical ecology of insects. Recent studies have identified chemoreceptors in coleopterans, facilitating the evolutionary analysis of not only ORs but also IRs and GRs. Thus, Cerambycidae, Tenebrionidae and Curculionidae have received increased attention. However, knowledge of the chemoreceptors from Scarabaeidae is still limited, particularly for those that are sympatric. Considering the roles of chemoreceptors, this analysis could shed light on evolutionary processes in the context of sympatry. Therefore, the aim of this study was to identify and compare the repertoires of ORs, GRs and IRs between two sympatric scarab beetles, Hylamorpha elegans and Brachysternus prasinus. Here, construction of the antennal transcriptomes of both scarab beetle species and analyses of their phylogeny, molecular evolution and relative expression were performed. Thus, 119 new candidate chemoreceptors were identified for the first time, including 17 transcripts for B. prasinus (1 GR, 3 IRs and 13 ORs) and 102 for H. elegans (22 GRs, 14 IRs and 66 ORs). Orthologs between the two scarab beetle species were found, revealing specific expansions as well as absence in some clades. Purifying selection appears to have occurred on H. elegans and B. prasinus ORs. Further efforts will be focused on target identification to characterize kairomone and/or pheromone receptors.
Asunto(s)
Escarabajos , Receptores Odorantes , Gorgojos , Animales , Transcriptoma , Simpatría , Perfilación de la Expresión Génica , Escarabajos/genética , Escarabajos/metabolismo , Gorgojos/genética , Filogenia , Receptores Odorantes/genética , Receptores Odorantes/metabolismo , Proteínas de Insectos/genética , Proteínas de Insectos/metabolismo , Antenas de Artrópodos/metabolismoRESUMEN
Gestational hypothyroidism may lead to preeclampsia development. However, this pathophysiological is unknown. We expect to find a shared mechanism by comparing hypothyroidism and preeclampsia. From our transcriptome data, we recognized olfactory receptors as that fingerprint. The reduction of taste and smell in hypothyroid patients has been known for a long time. Therefore, we decided to look to the olfactory receptors and aimed to identify genes capable of predicting preeclampsia (PEC). Methods: An Ion Proton Sequencer (Thermo Fisher Scientific, Waltham, MA, USA) was used to construct the transcriptome databases. RStudio with packages Limma v.3.50.0, GEOquery v.2.62.2, and umap v.0.2.8.8 were used to analyze the differentially expressed genes in GSE149440 from the Gene Expression Omnibus (GEO). The 7500 Real-Time PCR System (Applied Biosystems, Foster City, CA, USA) was used for RT-qPCR amplification of OR6X1 and OR4E2. Results: Our transcriptomic datasets analysis revealed 25.08% and 26.75% downregulated olfactory receptor (ORs) in mild nontreated gestational hypothyroidism (GHT) and PEC, respectively. In the GSE149440 GEO dataset, we found OR5H1, OR5T3, OR51A7, OR51B6, OR10J5, OR6C6, and OR2AG2 as predictors of early-onset PEC. We also evaluate two chosen biomarkers' responses to levothyroxine. The RT-qPCR demonstrated a difference in OR6X1 and OR4E2 expression between GHT and healthy pregnancy (p < 0.05). Those genes presented a negative correlation with TSH (r: -0.51, p < 0.05; and r: -0.44, p < 0.05), a strong positive correlation with each other (r: 0.89; p < 0.01) and the levothyroxine-treated group had no difference from the healthy one. We conclude that ORs could be used as biomarkers at the beginning of gestation, and the downregulated ORs found in GHT may be improved with levothyroxine treatment.
Asunto(s)
Ácidos Nucleicos Libres de Células , Hipotiroidismo , Preeclampsia , Receptores Odorantes , Embarazo , Femenino , Humanos , Preeclampsia/genética , Tiroxina , Receptores Odorantes/genética , Hipotiroidismo/genética , BiomarcadoresRESUMEN
Odorant receptors (ORs) are key specialized units for mate and host finding in moths of the Ditrysia clade, to which 98% of the lepidopteran species belong. Moth ORs have evolved to respond to long unsaturated acetates, alcohols, or aldehydes (Type I sex pheromones), falling into conserved clades of pheromone receptors (PRs). These PRs might have evolved from old lineages of non-Ditrysian moths that use plant volatile-like pheromones. However, a Ditrysian moth called the greater wax moth, Galleria mellonella (a worldwide-distributed pest of beehives), uses C9-C11 saturated aldehydes as the main sex pheromone components (i.e., nonanal and undecanal). Thus, these aldehydes represent unusual components compared with the majority of moth species that use, for instance, Type I sex pheromones. Current evidence shows a lack of consensus in the amount of ORs for G. mellonella, although consistent in that the moth does not have conserved PRs. Using genomic data, 62 OR candidates were identified, 16 being new genes. Phylogeny showed no presence of ORs in conserved PR clades. However, an OR with the highest transcript abundance, GmelOR4, appeared in a conserved plant volatile-detecting clade. Functional findings from the HEK system showed the OR as sensitive to nonanal and 2-phenylacetaldehyde, but not to undecanal. It is believed that to date GmelOR4 represents the first, but likely not unique, OR with a stable function in detecting aldehydes that help maintain the life cycle of G. mellonella around honey bee colonies.
Asunto(s)
Mariposas Nocturnas , Receptores Odorantes , Atractivos Sexuales , Animales , Abejas/genética , Mariposas Nocturnas/genética , Atractivos Sexuales/genética , Aldehídos , Receptores de Feromonas/genética , Receptores Odorantes/genéticaRESUMEN
Olfaction is a crucial capability for most vertebrates and is realized through olfactory receptors in the nasal cavity. The enormous diversity of olfactory receptors has been created by gene duplication, following a birth-and-death model of evolution. The olfactory receptor genes of the amphibians have received relatively little attention up to now, although recent studies have increased the number of species for which data are available. This study analyzed the diversity and chromosomal distribution of the OR genes of three anuran species (Engystomops pustulosus, Bufo bufo and Hymenochirus boettgeri). The OR genes were identified through searches for homologies, and sequence filtering and alignment using bioinformatic tools and scripts. A high diversity of OR genes was found in all three species, ranging from 917 in B. bufo to 1194 in H. boettgeri, and a total of 2076 OR genes in E. pustulosus. Six OR groups were recognized using an evolutionary gene tree analysis. While E. pustulosus has one of the highest numbers of genes of the gamma group (which detect airborne odorants) yet recorded in an anuran, B. bufo presented the smallest number of pseudogene sequences ever identified, with no pseudogenes in either the beta or epsilon groups. Although H. boettgeri shares many morphological adaptations for an aquatic lifestyle with Xenopus, and presented a similar number of genes related to the detection of water-soluble odorants, it had comparatively far fewer genes related to the detection of airborne odorants. This study is the first to describe the complete OR repertoire of the three study species and represents an important contribution to the understanding of the evolution and function of the sense of smell in vertebrates.
Asunto(s)
Receptores Odorantes , Animales , Filogenia , Receptores Odorantes/genética , Seudogenes/genética , Anuros/genética , Olfato/genéticaRESUMEN
The broad receptive field of the olfactory receptors constitutes the basis of a combinatorial code that allows animals to detect and discriminate many more odorants than the actual number of receptor types that they express. One drawback is that high odor concentrations recruit lower affinity receptors which can lead to the perception of qualitatively different odors. Here we addressed the contribution that signal-processing in the antennal lobe makes to reduce concentration dependence in odor representation. By means of calcium imaging and pharmacological approach we describe the contribution that GABA receptors play in terms of the amplitude and temporal profiles of the signals that convey odor information from the antennal lobes to higher brain centers. We found that GABA reduces the amplitude of odor elicited signals and the number of glomeruli that are recruited in an odor-concentration-dependent manner. Blocking GABA receptors decreases the correlation among glomerular activity patterns elicited by different concentrations of the same odor. In addition, we built a realistic mathematical model of the antennal lobe that was used to test the viability of the proposed mechanisms and to evaluate the processing properties of the AL network under conditions that cannot be achieved in physiology experiments. Interestingly, even though based on a rather simple topology and cell interactions solely mediated by GABAergic lateral inhibitions, the AL model reproduced key features of the AL response upon different odor concentrations and provides plausible solutions for concentration invariant recognition of odors by artificial sensors.
Asunto(s)
Neuronas Receptoras Olfatorias , Receptores Odorantes , Animales , Odorantes , Olfato/fisiología , Neuronas Receptoras Olfatorias/fisiología , Receptores Odorantes/metabolismo , Receptores de GABA , Vías Olfatorias/fisiologíaRESUMEN
Introducción: el papel clave del olfato, antiguo sistema sensorial, es proporcionar información sobre las sustancias químicas en el medio ambiente. El olfato desempeña un papel en la detección de compuestos peligrosos, el mantenimiento de la nutrición, el comportamiento interpersonal, la salud neurológica y la sensación de placer, entre otras funciones. En consecuencia, la disfunción olfativa puede conducir a un riesgo de lesiones, desnutrición, aislamiento social y una mala calidad de vida. Materiales y métodos: se realizó una exploración bibliográfica y se identificaron artículos de acuerdo con los criterios de inclusión y exclusión definidos y se tomaron aquellos con calidad en la evidencia. Discusión: el sistema olfativo humano tiene diferencias anatómicas, fisiológicas y genéticas considerables con respecto al de otros mamíferos. Conclusiones: las destrezas olfativas varían con factores como la edad, el sexo, la etapa de desarrollo, ciertas enfermedades otorrinolaringológicas y enfermedades generales.
Introduction: The key role of the ancient olfactory sensory system is to provide information about chemicals in the environment. Smell plays a role in the detection of dangerous compounds, the maintenance of nutrition, interpersonal behavior, neurological health, and the sensation of pleasure, among other functions. Consequently, olfactory dysfunction can lead to a risk of injury, malnutrition, social isolation, and a poor quality of life. Materials and methods: A bibliographical exploration was carried out and articles were identified according to the inclusion and exclusion criteria defined and those with quality evidence were taken. Discussion: The human olfactory system has considerable anatomical, physiological, and genetic differences from that of other mammals. Conclusions: Olfactory skills vary with factors such as age, sex, stage of development, certain ear, nose and throat diseases and general diseases.
Asunto(s)
Humanos , Masculino , Femenino , Olfato , Otorrinolaringólogos , Nervio Olfatorio , Receptores OdorantesRESUMEN
The pepper weevil Anthonomus eugenii is one of the most damaging pests to the pepper crop. To offer alternative management strategies to insecticides, several studies have identified the semiochemicals that are involved in the pepper weevil's aggregation and mating behavior; however, there is no information on its perireceptor molecular mechanism, to date. In this study, bioinformatics tools were used to functionally annotate and characterize the A. eugenii head transcriptome and their probable coding proteins. We identified twenty-two transcripts belonging to families related to chemosensory processes, seventeen corresponding to odorant-binding proteins (OBP), and six to chemosensory proteins (CSP). All results matched with closely related Coleoptera: Curculionidae homologous proteins. Likewise, twelve OBP and three CSP transcripts were experimentally characterized by RT-PCR in different female and male tissues. The results by sex and tissue display the different expression patterns of the AeugOBPs and AeugCSPs; some are present in both sexes and all tissues, while others show expressions with higher specificity, which suggests diverse physiological functions in addition to chemo-detection. This study provides information to support the understanding of odor perception in the pepper weevil.
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Escarabajos , Insecticidas , Receptores Odorantes , Gorgojos , Humanos , Animales , Masculino , Femenino , Gorgojos/genética , Escarabajos/genética , Odorantes , Transcriptoma , Proteínas de Insectos/genética , Receptores Odorantes/metabolismo , Filogenia , Perfilación de la Expresión GénicaRESUMEN
Odorant receptors (ORs) belong to a large family of G protein-coupled receptors (GPCRs) that are highly expressed by olfactory sensory neurons of the nose. Accumulating evidence indicates that they are also expressed in a variety of nonolfactory tissues, which makes them new potential drug targets. Here we discuss the challenges and strategies to target these receptors.
Asunto(s)
Neuronas Receptoras Olfatorias , Receptores Odorantes , Humanos , Receptores Acoplados a Proteínas GRESUMEN
Environmental enrichment techniques include olfactory stimuli for improving animal welfare. This study aimed to analyze the reactions of 41 shelter dogs exposed to odorous stimuli, such as the method used in another study on wild canids. The focal animal method analyzed the dogs' reactions, with all behaviors recorded. Behavioral responses were classified as positive (P+), negative (N-), or other (Ot). Independent variables were all dogs and the size of the packs. The behavior between the basal (without stimulus), exposure, and after-stimulus withdrawal was analyzed. For all dogs, olfactory stimuli significantly increased P+ (P=0.001) and N- (P=0.004), contrasting with the decrement of Ot behaviors (P=0.001) from the basal to the exposure phase. After the withdrawal of the stimuli, P+, N-, and Ot behaviors returned to basal levels (P>0.05). There were no significant differences (P>0.05) in the conduct of small or large packs exposed to stimuli. Dogs are sensitive to olfactory stimuli, but arousal is generalized to P+ and N-. It is undesirable to an N- increase for improvement of animal welfare. Contrary to what was observed in a study with wild canids, the method failed in shelter dogs because N- was increased. The introduction of sudden novelty (olfactory stimulus) in an impoverished shelter environment may have caused excitement in the dogs. It is suggested that changes in the method, such as stimuli exposition to each dog in an isolated room, are necessary to increase sheltered dog well-being.(AU)
As técnicas de enriquecimento ambiental incluem estímulos olfativos para aumentar o bem-estar animal. O objetivo deste estudo foi analisar as reações de 41 cães de abrigo expostos a estímulos odoríferos, como o método utilizado em outro estudo com canídeos selvagens. As reações dos cães foram analisadas pelo método animal focal, com todos os comportamentos registrados. As respostas comportamentais foram classificadas como positivas (P+), negativas (N-) ou outras (Ot). As variáveis independentes foram todos os cães e o tamanho das matilhas. Foi analisado o comportamento entre o basal (sem estímulo), exposição e após a retirada do estímulo. Para todos os cães, os estímulos olfativos aumentaram significativamente P+ (P=0,001) e N- (P=0,004), contrastando com a diminuição dos comportamentos Ot (P=0,001) da fase basal para a de exposição. Após a retirada dos estímulos, os comportamentos P+, N- e Ot retornaram aos níveis basais (P>0,05). Não houve diferenças significativas (P>0,05) no comportamento de matilhas pequenas ou grandes expostas a estímulos. Os cães são sensíveis a estímulos olfativos, mas a excitação parece ser generalizada para ambos, P+ e N-. É indesejável um aumento de N- para melhoria do bem-estar animal. Ao contrário do que foi observado em um estudo com canídeos selvagens, o método falhou em abrigar cães porque o N- foi aumentado. A introdução de uma novidade repentina (estímulo olfativo) em um ambiente de abrigo empobrecido, pode ter causado excitação exagerada nos cães. Sugere-se alterações no método, como a exposição de estímulos a cada cão em uma sala isolada necessária para aumentar o bem-estar do cão abrigado.(AU)
Asunto(s)
Animales , Receptores Odorantes/análisis , Perros/anatomía & histología , Percepción Olfatoria/fisiología , Bienestar del Animal , Vivienda para AnimalesRESUMEN
INTRODUCTION AND OBJECTIVES: Obesity is a global health problem that triggers fat liver accumulation. The prevalence of obesity and the risk of non-alcoholic steatohepatitis (NASH) among young obese Mexican is high. Furthermore, genetic predisposition is a key factor in weight gain and disrupts metabolism. Herein, we used Whole-Exome Sequencing to identify potential causal variants and the biological processes that lead to obesity with progression to NASH among Mexican patients. MATERIALS AND METHODS: Whole-Exome Sequencing was performed in nine obese patients with NASH diagnosis with a BMI ≥30 kg/m2 and one control (BMI=24.2 kg/m2) by using the Ion S5TM platform. Genetic variants were determined by Ion Reporter software. Enriched GO biological set genes were identified by the WebGestalt tool. Genetic variants within ≥2 obese NASH patients and having scores of SIFT 0.0-0.05 and Polyphen 0.85-1.0 were categorized as pathogenic. RESULTS: A total of 1359 variants with a probable pathogenic effect were determined in obese patients with NASH diagnosis. After several filtering steps, the most frequent pathogenic variants found were rs25640-HSD17B4, rs8105737-OR1I1, rs998544-OR5R1, and rs4916685, rs10037067, and rs2366926 in ADGRV1. Notably, the primary biological processes affected by these pathogenic variants were the sensory perception and detection of chemical stimulus pathways in which the olfactory receptor gene family was the most enriched. CONCLUSIONS: Variants in the olfactory receptor genes were highly enriched in Mexican obese patients that progress to NASH and could be potential targets of association studies.
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Enfermedad del Hígado Graso no Alcohólico , Receptores Odorantes , Humanos , Enfermedad del Hígado Graso no Alcohólico/diagnóstico , Enfermedad del Hígado Graso no Alcohólico/epidemiología , Enfermedad del Hígado Graso no Alcohólico/genética , Receptores Odorantes/genética , Receptores Odorantes/metabolismo , Secuenciación del Exoma , Hígado/patología , Obesidad/diagnóstico , Obesidad/epidemiología , Obesidad/genéticaRESUMEN
The Aedes aegypti mosquito is the main hematophagous vector responsible for arbovirus transmission in Brazil. The disruption of A. aegypti hematophagy remains one of the most efficient and least toxic methods against these diseases and, therefore, efforts in the research of new chemical entities with repellent activity have advanced due to the elucidation of the functionality of the olfactory receptors and the behavior of mosquitoes. With the growing interest of the pharmaceutical and cosmetic industries in the development of chemical entities with repellent activity, computational studies (e.g., virtual screening and molecular modeling) are a way to prioritize potential modulators with stereoelectronic characteristics (e.g., pharmacophore models) and binding affinity to the AaegOBP1 binding site (e.g., molecular docking) at a lower computational cost. Thus, pharmacophore- and docking-based virtual screening was employed to prioritize compounds from Sigma-Aldrich® (n = 126,851) and biogenic databases (n = 8766). In addition, molecular dynamics (MD) was performed to prioritize the most potential potent compounds compared to DEET according to free binding energy calculations. Two compounds showed adequate stereoelectronic requirements (QFIT > 81.53), AaegOBP1 binding site score (Score > 42.0), volatility and non-toxic properties and better binding free energy value (∆G < −24.13 kcal/mol) compared to DEET ((N,N-diethyl-meta-toluamide)) (∆G = −24.13 kcal/mol).
Asunto(s)
Aedes , Repelentes de Insectos , Receptores Odorantes , Animales , Receptores Odorantes/metabolismo , DEET/química , Simulación del Acoplamiento Molecular , Mosquitos Vectores , Repelentes de Insectos/farmacología , Repelentes de Insectos/química , Preparaciones Farmacéuticas/metabolismoRESUMEN
Insect chemosensory systems, such as smell and taste, are mediated by chemosensory receptor and non-receptor protein families. In the last decade, many studies have focused on discovering these families in Tephritidae species of agricultural importance. However, to date, there is no information on the Mexican fruit fly Anastrepha ludens Loew, a priority pest of quarantine importance in Mexico and other countries. This work represents the first effort to identify, classify and characterize the six chemosensory gene families by analyzing two head transcriptomes of sexually immature and mature adults of A. ludens from laboratory-reared and wild populations, respectively. We identified 120 chemosensory genes encoding 31 Odorant-Binding Proteins (OBPs), 5 Chemosensory Proteins (CSPs), 2 Sensory Neuron Membrane Proteins (SNMPs), 42 Odorant Receptors (ORs), 17 Ionotropic Receptors (IRs), and 23 Gustatory Receptors (GRs). The 120 described chemosensory proteins of the Mexican fruit fly significantly contribute to the genetic databases of insects, particularly dipterans. Except for some OBPs, this work reports for the first time the repertoire of olfactory proteins for one species of the genus Anastrepha, which provides a further basis for studying the olfactory system in the family Tephritidae, one of the most important for its economic and social impact worldwide.
Asunto(s)
Receptores Odorantes , Tephritidae , Animales , Perfilación de la Expresión Génica , Proteínas de Insectos/genética , Proteínas de Insectos/metabolismo , Receptores Odorantes/genética , Receptores Odorantes/metabolismo , Olfato , Tephritidae/genética , Tephritidae/metabolismo , TranscriptomaRESUMEN
Arboviruses are a group of viruses (e.g. Dengue, Chikungunya and Yellow fever virus) that are transmitted by arthropod vectors, which Aedes aegipty is the vector of main viruses in Americas. This vector is responsible to 2.4 millions of arboviruses cases in Brazil with less than a thousand deaths annually. Despite of epidemiological data, arboviruses treatment is symptomatic and the vaccine control is not effective, which makes the vector control against A. aegipty a promising strategy to diseases control. One way to achieve this goal is to development of A. aegipty sensitive olfactory modulators. Odorant binding protein 1 from A. aegypti (AaegOBP1) is essential in sensory communication, and is the first filter in odorant selection, which makes this target promising to development of new repellents. For this reason, hierarchical virtual screening (ligand-based pharmacophore model and molecular docking) together volatility filter was applied at Sigma-Aldrich database (n = 126.851) to prioritize potential molecules to repellency assays. Three compounds showed adequate stereo-electronic requirements (QFIT> 81.53), score to AaegOBP1 binding site (Score > 36.0) and volatile properties and it was chosen for repellency assays. ZINC00170981 and ZINC00131924 showed a dose-response behavior, while ZINC01621824 did not showed activity in repellency assays. Finally, Molecular Dynamics (MD) was employed to hypothesize the stability of protein-ligand complexes. According to RMSD, RMSF and binding free energy data, ZINC00170981 and ZINC00131924 were able to stabilize AaegOBP1 binding-site during the trajectory by interactions with key residues such as His77, Leu89 and Trp114). Communicated by Ramaswamy H. Sarma.
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Aedes , Animales , Bioensayo , Ligandos , Simulación del Acoplamiento Molecular , Mosquitos Vectores , Receptores OdorantesRESUMEN
O objetivo desse trabalho foi identificar as consequências moleculares e funcionais da falta da proteína Ric8b no epitélio olfatório de camundongos. Para esse fim, comparamos o transcriptoma de epitélio olfatório de camundongos knock-out tecido específico para a proteína RIC8B (Ric8b cKO) com o dos seus irmãos tipo selvagem (WT). Identificamos muitos genes que apresentaram expressão reduzida no epitélio olfatório do camundongo Ric8b cKO, mas também vários genes que apresentaram a sua expressão aumentada. A maioria dos genes com expressão reduzida corresponde a genes normalmente expressos em neurônios olfatórios maduros, como por exemplo os genes de receptores olfatórios, o que é compatível com o fato já conhecido de que os camundongos Ric8b cKO apresentam um menor número desses neurônios. Inesperadamente, apesar de a maioria dos genes de receptores olfatórios ter a sua expressão diminuída no camundongo Ric8b cKO, observamos que um grupo destes genes de receptores teve a sua expressão aumentada. Os camundongos Ric8b cKO apresentaram também genes marcadores de outros tipos celulares que não neurônios canônicos com expressão aumentada no seu epitélio olfatório. Dentre eles, os mais significativamente alterados foram os genes marcadores de neurônios Trpc2+ tipo B (que expressam a guanilato ciclase solúvel Gucy1b2). Sabe-se que este tipo de neurônio é responsável pela sensibilidade a diferentes gases, e concordantemente, observamos que os camundongos Ric8b cKO apresentaram um aumento da sensibilidade a gás carbônico. Como o olfato apresenta um papel importante na regulação de ingestão alimentar, analisamos como os camundongos Ric8b cKO se comportam frente a diferentes dietas. Interessantemente, observamos que esses animais não apresentam preferência por alimento rico em gorduras quando comparado aos seus irmãos tipo selvagem. Nossos resultados sugerem, portanto, que a ausência da proteína RIC8B resulta na alteração de representatividade de neurônios canônicos e não canônicos no epitélio olfatório de camundongos, o que por sua vez leva a alterações funcionais e comportamentais
The objective of this work was to identify the molecular and functional consequences of the lack of the RIC8B protein in the main olfactory epithelium of mice. To this end, we compared the olfactory epithelium transcriptome of Ric8b tissue-specific knock-out mice (Ric8b cKO) with that of their wild-type littermates (WT). We identified many genes with differential expression, many of which were downregulated and also some which were upregulated in the olfactory epithelium of the Ric8b cKO mice. Most of the downregulated genes correspond to genes normally expressed in mature olfactory sensory neurons, such as olfactory receptor genes. This is compatible with the already known fact that the Ric8b cKO mice have less of this kind of neuron. Unexpectedly, even though most of the olfactory receptor genes were downregulated, we observed a subset of these genes that had their expression upregulated in the Ric8b cKO mice. The Ric8b cKO mice also showed upregulation for genes that are markers for cell types other than canonic neurons in their olfactory epithelium. Among these, the most significantly altered were the markers for neurons Trpc2+ type B (that express the soluble guanylate cyclase Gucy1b2). It is known that this kind of neuron is responsible for sensitivity to different gases. Accordingly, we observed that the Ric8b cKO mice presented a higher sensitivity to carbon dioxide. Since olfaction has an important role in food intake, we analyzed how the Ric8b cKO mice behaved with different diets. Interestingly, we observed that the Ric8b cKO mice lack preference for high fat diet when compared to their wild-type littermates. Our results indicate, therefore, that the lack of the RIC8B protein results in altered representativity of canonic and non-canonic neurons in the olfactory epithelium of mice, which then leads to altered function and behavior
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Animales , Masculino , Femenino , Ratones , Mucosa Olfatoria/anomalías , Receptores Odorantes/agonistas , Neuronas Receptoras Olfatorias , Ratones Noqueados , Conducta Alimentaria/clasificación , Neuronas/química , AbsentismoRESUMEN
The communication and reproduction of insects are driven by chemical sensing. During this process, chemical compounds are transported across the sensillum lymph to the sensory neurons assisted by different types of soluble binding proteins: odorant-binding proteins (OBPs); chemosensory proteins (CSPs); some members of ML-family proteins (MD-2 (myeloid differentiation factor-2)-related Lipid-recognition), also known as NPC2-like proteins. Potential transcripts involved in chemosensing were identified by an in silico analysis of whole-body female and male transcriptomes of the parasitic wasp Diachasmimorpha longicaudata. This analysis facilitated the characterization of fourteen OBPs (all belonging to the Classic type), seven CSPs (and two possible isoforms), and four NPC2-like proteins. A differential expression analysis by qPCR showed that eleven of these proteins (CSPs 2 and 8, OBPs 2, 3, 4, 5, 6, 9, 10, and 11, and NPC2b) were over-expressed in female antenna and two (CSP 1 and OBP 12) in the body without antennae. Foraging behavior trials (linked to RNA interference) suggest that OBPs 9, 10, and 11 are potentially involved in the female orientation to chemical cues associated with the host. OBP 12 seems to be related to physiological processes of female longevity regulation. In addition, transcriptional silencing of CSP 3 showed that this protein is potentially associated with the regulation of foraging behavior. This study supports the hypothesis that soluble binding proteins are potentially linked to fundamental physiological processes and behaviors in D. longicaudata. The results obtained here contribute useful information to increase the parasitoid performance as a biological control agent of fruit fly pest species.
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Proteínas de Insectos/metabolismo , Receptores Odorantes/metabolismo , Avispas/metabolismo , Animales , Conducta Alimentaria , Femenino , Proteínas de Insectos/química , Proteínas de Insectos/genética , Masculino , Receptores Odorantes/química , Receptores Odorantes/genética , Transcriptoma , Avispas/genética , Avispas/fisiologíaRESUMEN
The evolution of obligate ectoparasitism in blowflies (Diptera: Calliphoridae) has intrigued scientists for over a century, and surprisingly, the genetics underlying this lifestyle remain largely unknown. Blowflies use odors to locate food and oviposition sites; therefore, olfaction might have played a central role in niche specialization within the group. In insects, the coreceptor Orco is a required partner for all odorant receptors (ORs), a major gene family involved in olfactory-evoked behaviors. Hence, we characterized the Orco gene in the New World screwworm, Cochliomyia hominivorax, a blowfly that is an obligate ectoparasite of warm-blooded animals. In contrast, most of the closely related blowflies are scavengers that lay their eggs on dead animals. We show that the screwworm Orco orthologue (ChomOrco) is highly conserved within Diptera, showing signals of strong purifying selection. Expression of ChomOrco is broadly detectable in chemosensory appendages, and is related to morphological, developmental, and behavioral aspects of the screwworm biology. We used CRISPR/Cas9 to disrupt ChomOrco and evaluate the consequences of losing the OR function on screwworm behavior. In two-choice assays, Orco mutants displayed an impaired response to floral-like and animal host-associated odors, suggesting that OR-mediated olfaction is involved in foraging and host-seeking behaviors in C. hominivorax. These results broaden our understanding of the chemoreception basis of niche occupancy by blowflies.
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Dípteros/fisiología , Conducta Alimentaria , Conducta de Búsqueda de Hospedador , Proteínas de Insectos/metabolismo , Receptores Odorantes/metabolismo , Animales , Dípteros/metabolismo , Proteínas de Insectos/genética , Mutación , Filogenia , Receptores Odorantes/genética , OlfatoRESUMEN
Malaria is a protozoan infection transmitted by the bite of the infected female mosquito belonging to the genus Anopheles spp., which causes more than 445 million annual deaths worldwide. Available drugs have serious adverse effects (e.g. blurred vision, hypotension and headache) and species-dependent efficacy. An alternative to overcome these problems involve the use of molecules with affinity to the Anopheles gambiae mosquito odor receptors, minimizing the reinfection process as well as reducing the problems related to pharmacological therapy. The vector control can interrupt the epidemiological cycle and, therefore, control the malaria incidence. In the olfactory pathway, odorant binding protein 1 acts on the first level of odor recognition on malarial vector and thus can be used to modulate mosquito behavior and development of new attracts or repellents. Thus, this study applied ligand-based (2D-chemical similarity) and structure-based (docking and molecular dynamics) computational approaches to prioritize potential olfactory modulators on natural products catalogs at ZINC15 database (n = 98,379). Hierarchical virtual screening prioritized a potential olfactory modulator (Z8217) against Anopheles gambiae odorant binding protein 1 (AgOBP1). Next, it was submitted to molecular dynamics routine to identify structural requirements and the interactions profile required for binding-site affinity. This promising natural compound can interact like experimental ligand and will be used in repellency assay to confirm its sensorial behavior.Communicated by Ramaswamy H. Sarma.