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1.
Int J Mol Sci ; 24(2)2023 Jan 13.
Artigo em Inglês | MEDLINE | ID: mdl-36675132

RESUMO

Insects use a powerful and complex olfactory recognition system to sense odor molecules in the external environment to guide behavior. A large family of odorant receptors (ORs) mediates the detection of pheromone compounds. Anoplophora glabripennis is a destructive pest that harms broad-leaved tree species. Although olfactory sensation is an important factor affecting the information exchange of A. glabripennis, little is known about the key ORs involved. Here, we identified ninety-eight AglaORs in the Agla2.0 genome and found that the AglaOR gene family had expanded with structural and functional diversity. RT-qPCR was used to analyze the expression of AglaORs in sex tissues and in adults at different developmental stages. Twenty-three AglaORs with antennal-biased expression were identified. Among these, eleven were male-biased and two were female-biased and were more significantly expressed in the sexual maturation stage than in the post-mating stage, suggesting that these genes play a role in sexual communication. Relatively, two female-biased AglaORs were overexpressed in females seeking spawning grounds after mating, indicating that these genes might be involved in the recognition of host plant volatiles that may regulate the selection of spawning grounds. Our study provides a theoretical basis for further studies into the molecular mechanism of A. glabripennis olfaction.


Assuntos
Besouros , Receptores Odorantes , Animais , Feminino , Masculino , Receptores Odorantes/genética , Receptores Odorantes/metabolismo , Besouros/metabolismo , Olfato , Plantas/metabolismo , Comunicação , Proteínas de Insetos/genética , Proteínas de Insetos/metabolismo , Antenas de Artrópodes/metabolismo
2.
Int J Mol Sci ; 24(2)2023 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-36675226

RESUMO

Euplatypus parallelus is one of the dominant rubber bark beetle species in Hainan's rubber-planting area. Semiochemicals, including the volatiles found in rubber trees and aggregation pheromones, play an important role in the search for suitable host plants. To examine the possible functional role of highly expressed odorant-binding protein 2 of Euplatypus parallelus (EparOBP2) in the semiochemical recognition process, we cloned and analyzed the cDNA sequence of EparOBP2. The results showed that EparOBP2 contains an open reading frame (ORF) of 393 bp that encodes 130 amino acids, including a 21-amino-acid residue signal peptide at the N-terminus. The matured EparOBP2 protein consists of seven α-helices, creating an open binding pocket and three disulfide bridges. The results of the fluorescence binding assay showed that EparOBP2 had high binding ability with α-pinene and myrcene. The docking results confirmed that the interactions of α-pinene and myrcene with EparOBP2 were primarily achieved through hydrophobic interactions. This study provides evidence that EparOBP2 may be involved in the chemoreception of semiochemicals and that it can successfully contribute to the integrated management of E. parallelus.


Assuntos
Receptores Odorantes , Gorgulhos , Animais , Feromônios/metabolismo , Borracha , Gorgulhos/metabolismo , Receptores Odorantes/metabolismo , Proteínas de Insetos/metabolismo , Ligação Proteica
3.
Chem Senses ; 482023 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-36625229

RESUMO

Humans have significant individual variations in odor perception, derived from their experience or sometimes from differences in the olfactory receptor (OR) gene repertoire. In several cases, the genetic variation of a single OR affects the perception of its cognate odor ligand. Musks are widely used for fragrance and are known to demonstrate specific anosmia. It, however, remains to be elucidated whether the OR polymorphism contributes to individual variations in musk odor perception. Previous studies reported that responses of the human musk receptor OR5AN1 to a variety of musks in vitro correlated well with perceptual sensitivity to those odors in humans and that the mouse ortholog, Olfr1440 (MOR215-1), plays a critical role in muscone perception. Here, we took advantage of genetic variation in OR5AN1 to examine how changes in receptor sensitivity are associated with human musk perception. We investigated the functional differences between OR5AN1 variants in an in vitro assay and measured both perceived intensity and detection threshold in human subjects with different OR5AN1 genotypes. Human subjects homozygous for the more sensitive L289F allele had a lower detection threshold for muscone and found macrocyclic musks to be more intense than subjects homozygous for the reference allele. These results demonstrate that the genetic variation in OR5AN1 contributes to perceptual differences for some musks. In addition, we found that the more functional variant of OR5A1, a receptor involved in ß-ionone perception, is associated with the less functional variant of OR5AN1, suggesting that the perceived intensities of macrocyclic musks and ß-ionone are inversely correlated.


Assuntos
Percepção Olfatória , Receptores Odorantes , Humanos , Camundongos , Animais , Receptores Odorantes/genética , Odorantes , Variação Genética , Percepção , Percepção Olfatória/genética , Receptores Colinérgicos/genética , Receptores Proteína Tirosina Quinases/genética
4.
J Agric Food Chem ; 71(2): 1067-1076, 2023 Jan 18.
Artigo em Inglês | MEDLINE | ID: mdl-36598383

RESUMO

A total of 12 OBPs were identified in the antennae of ladybird Hippodamia variegata. HvarOBP1/2/5/6/10/11 were highly expressed in adults, whereas HvarOBP3/4/7/8/9/12 had higher expression levels in larvae. In adults, HvarOBP1/2/4/5/6/7/8/11/12 were highly expressed in antennae. Among these, recombinant HvarOBP5 strong bound with (E)-ß-farnesene (EßF), (R)-(+)-limonene, (E,E)-4,8,12-trimethyltrideca-l,3,7,11-tetraene (TMTT), (E)-4,8-dimethyl-1,3,7-nonatriene (DMNT), hexyl hexanoate, and geranyl acetate. Molecular docking indicated that Leu42, Lys43, and His64 were the key binding sites of HvarOBP5. All six ligands evoked electroantennography (EAG) responses in ladybirds. Moreover, (R)-(+)-limonene and hexyl hexanoate were attractive to both sexes. After RNA interference for 72 h, the EAGs of dsRNA-injected ladybirds to DMNT and hexyl hexanoate were significantly decreased by 73.8 and 78.6%, respectively. Both dsRNA-injected males and females showed significantly lower behavioral preferences for DMNT and hexyl hexanoate. These findings suggest that HvarOBP5 in H. variegata plays an important role in the perception of semiochemical cues from preys and habitat plants.


Assuntos
Feromônios , Receptores Odorantes , Feminino , Masculino , Animais , Feromônios/metabolismo , Caproatos , Limoneno , Simulação de Acoplamento Molecular , Receptores Odorantes/metabolismo , Percepção , Proteínas de Insetos/genética , Antenas de Artrópodes/metabolismo
5.
J Agric Food Chem ; 71(4): 1837-1844, 2023 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-36682010

RESUMO

Odorant receptors (ORs) in insects are crucial for the detection of chemical signals. However, the functions of the conserved OR genes among insect species are rarely studied. In this study, we analyzed a well-conserved OR clade in Diptera insects and cloned a gene from this clade, EcorOR4, in the hoverfly Eupeodes corollae. Real-time quantitative PCR showed that EcorOR4 was highly expressed in the antennae and upregulated in the mated females, and in vitro functional characterization showed that EcorOR4 was narrowly tuned to 1-octen-3-ol. Electroantennogram assays revealed that the antennal response of mated females to 1-octen-3-ol was significantly higher than that of mated males, but no significant differences were observed between male and female virgins. Finally, a Y-tube olfactometer bioassay showed that 1-octen-3-ol is an attractant for only mated female E. corollae adults. These results demonstrate that EcorOR4 is involved in the detection of 1-octen-3-ol and that this compound may affect the host-finding and oviposition behavior in female E. corollae.


Assuntos
Dípteros , Receptores Odorantes , Animais , Feminino , Masculino , Receptores Odorantes/genética , Dípteros/genética , Octanóis , Oviposição
6.
Sci Rep ; 13(1): 971, 2023 Jan 18.
Artigo em Inglês | MEDLINE | ID: mdl-36653421

RESUMO

Odorants are detected by olfactory sensory neurons, which are covered by olfactory mucus. Despite the existence of studies on olfactory mucus, its constituents, functions, and interindividual variability remain poorly understood. Here, we describe a human study that combined the collection of olfactory mucus and olfactory psychophysical tests. Our analyses revealed that olfactory mucus contains high concentrations of solutes, such as total proteins, inorganic elements, and molecules for xenobiotic metabolism. The high concentrations result in a capacity to capture or metabolize a specific repertoire of odorants. We provide evidence that odorant metabolism modifies our sense of smell. Finally, the amount of olfactory mucus decreases in an age-dependent manner. A follow-up experiment recapitulated the importance of the amount of mucus in the sensitive detection of odorants by their receptors. These findings provide a comprehensive picture of the molecular processes in olfactory mucus and propose a potential cause of olfactory decline.


Assuntos
Neurônios Receptores Olfatórios , Receptores Odorantes , Humanos , Olfato/fisiologia , Receptores Odorantes/metabolismo , Neurônios Receptores Olfatórios/metabolismo , Odorantes/análise , Muco/metabolismo
7.
Int J Mol Sci ; 24(2)2023 Jan 14.
Artigo em Inglês | MEDLINE | ID: mdl-36675176

RESUMO

Heparin, a class of glycosaminoglycans (GAGs), is widely used to induce sperm capacitation and fertilization. How heparin induces sperm capacitation remains unclear. Olfactory receptors (ORs) which are G protein-coupled receptors, have been proposed to be involved in sperm capacitation. However, the interaction between ORs and odor molecules and the molecular mechanism of ORs mediating sperm capacitation are still unclear. The present study aimed to explore the underlying interaction and mechanism between heparin and ORs in carrying out the boar sperm capacitation. The results showed that olfactory receptor 2C1 (OR2C1) is a compulsory unit which regulates the sperm capacitation by recognizing and binding with heparin, as determined by Dual-Glo Luciferase Assay and molecular docking. In addition, molecular dynamics (MD) simulation indicated that OR2C1 binds with heparin via a hydrophobic cavity comprises of Arg3, Ala6, Thr7, Asn171, Arg172, Arg173, and Pro287. Furthermore, we demonstrated that knocking down OR2C1 significantly inhibits sperm capacitation. In conclusion, we highlighted a novel olfactory receptor, OR2C1, in boar sperm and disclosed the potential binding of heparin to Pro287, a conserved residue in the transmembrane helices region 7 (TMH7). Our findings will benefit the further understanding of ORs involved in sperm capacitation and fertilization.


Assuntos
Heparina , Receptores Odorantes , Capacitação Espermática , Animais , Masculino , Heparina/farmacologia , Heparina/metabolismo , Simulação de Acoplamento Molecular , Receptores Odorantes/genética , Receptores Odorantes/metabolismo , Sêmen/metabolismo , Capacitação Espermática/genética , Capacitação Espermática/fisiologia , Espermatozoides/metabolismo , Suínos
8.
Biosens Bioelectron ; 224: 115047, 2023 Mar 15.
Artigo em Inglês | MEDLINE | ID: mdl-36628827

RESUMO

Gas sensing based on graphene field-effect transistors (GFETs) has gained broad interest due to their high sensitivity. Further progress in gas sensing with GFETs requires to detection of various odor molecules for applications in the environmental monitoring, healthcare, food, and cosmetic industries. To develop the ubiquitous odor-sensing system, establishing an artificial sense of smell with electronic devices by mimicking olfactory receptors will be key. Although the application of olfactory receptors to GFETs is straightforward for odor sensing, synthetic molecules with a similar function to olfactory receptors would be desirable to realize the robust performance of sensing. In this work, we designed three new peptides consisting of two domains: a bio-probe to the target molecules and a molecular scaffold. These peptides were rationally designed based on a motif sequence in olfactory receptors and self-assembled into a molecular thin film on GFETs. Limonene, methyl salicylate, and menthol were employed as representative odor molecules of plant flavors to demonstrate the biosensing of odor molecules. The conductivity change of GFETs against the binding to odor molecules with various concentrations and the dynamic response revealed a distinct signature of three different peptides against individual species of the target molecules. The kinetic response of each peptide exhibited characteristic time constants in the adsorption and desorption process, also supported by the principal component analysis. Our demonstration of the graphene odor sensors with the designed peptides opens a way to establish future peptide-array sensors with multi-sequence of peptide, realizing an odor sensing system with higher selectivity.


Assuntos
Técnicas Biossensoriais , Grafite , Receptores Odorantes , Odorantes , Grafite/química , Transistores Eletrônicos , Peptídeos
9.
J Comp Neurol ; 531(1): 116-131, 2023 01.
Artigo em Inglês | MEDLINE | ID: mdl-36161277

RESUMO

Lungfish are the fish related most closely to tetrapods. The olfactory organ of lungfish contains two distinct sensory epithelia: the lamellar olfactory epithelium (OE) and the recess epithelium (RecE). Based on their ultrastructural and histological characteristics, the lamellar OE and the RecE are considered to correspond respectively to the teleost OE and a primitive vomeronasal organ (VNO). In tetrapods, the OE and VNO have been shown to express different families of olfactory receptors; for example, in mammals, the OE expresses odorant receptors and trace amine-associated receptors, while the VNO expresses type 1 (V1Rs) and type 2 (V2Rs) vomeronasal receptors. In the present study, we examined the expression of V1Rs in the olfactory organs of two African lungfish, Protopterus annectens and Protopterus amphibius. RNA sequencing and phylogenetic analyses identified 29 V1R genes in P. annectens and 50 V1R genes in P. amphibius. Most V1Rs identified in these lungfish were classified as the tetrapod-type V1Rs initially found in tetrapods and distinct from fish-type V1Rs. In teleost, which all lack a VNO, all olfactory receptors are expressed in the OE, while in Xenopus V1Rs are expressed exclusively in the OE, and not in the VNO. In situ hybridization analysis indicated that lungfish V1Rs were expressed mainly in the lamellar OE and rarely in the RecE. These results imply that V1R expression in lungfish represents an intermediate step toward the complete segregation of V1R expression between the OE and VNO, reflecting the phylogenetic position of lungfish between teleosts and amphibians.


Assuntos
Neurônios Receptores Olfatórios , Receptores Odorantes , Órgão Vomeronasal , Animais , Receptores Odorantes/genética , Filogenia , Órgão Vomeronasal/metabolismo , Mucosa Olfatória/metabolismo , Neurônios Receptores Olfatórios/metabolismo , Peixes , Mamíferos
10.
Biosens Bioelectron ; 223: 115034, 2023 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-36574741

RESUMO

The ectopic co-expression of taste and olfactory receptors in cardiomyocytes provides not only possibilities for the construction of biomimetic gustatory and olfactory sensors but also promising novel therapeutic targets for tachycardia treatment. Here, bitter taste and olfactory receptors endogenously expressed in HL-1 cells were verified by RT-PCR and immunofluorescence staining. Then HL-1 cardiomyocyte-based integrated gustatory and olfactory sensing array coupling with the microelectrode array (MEA) was first constructed for drugs screening and evaluation for tachycardia treatment. The MEA sensor detected the extracellular field potentials and reflected the systolic-diastolic properties of cardiomyocytes in real time in a label-free and non-invasive way. The in vitro tachycardia model was constructed using isoproterenol as the stimulator. The proposed sensing array facilitated potential drug screening for tachycardia treatment, such as salicin, artemisinin, xanthotoxin, and azelaic acid which all activated specific receptors on HL-1 cells. IC50 values for four potential drugs were calculated to be 0.0036 µM, 309.8 µM, 14.68 µM, and 0.102 µM, respectively. Visualization analysis with heatmaps and PCA cluster showed that different taste and odorous drugs could be easily distinguished. The mean inter-class Euclidean distance between different bitter drugs was 1.681, which was smaller than the distance between bitter and odorous drugs of 2.764. And the inter-class distance was significantly higher than the mean intra-class Euclidean distance of 1.172. In summary, this study not only indicates a new path for constructing novel integrated gustatory and olfactory sensors but also provides a powerful tool for the quantitative evaluation of potential drugs for tachycardia treatment.


Assuntos
Técnicas Biossensoriais , Receptores Odorantes , Humanos , Miócitos Cardíacos , Avaliação Pré-Clínica de Medicamentos , Biomimética , Olfato , Paladar , Taquicardia
11.
J Adv Res ; 43: 1-12, 2023 01.
Artigo em Inglês | MEDLINE | ID: mdl-36585100

RESUMO

INTRODUCTION: The parasitoid wasp Microplitis mediator is an important natural enemy of the turnip moth Agrotis segetum and other Noctuidae pests. In our field observation, it was fortuitously discovered that sex pheromone traps used for A. segetum also attract female wasps, verified by a simulated field condition dual-choice laboratory assay. Therefore, it was hypothesized that olfactory recognition could be crucial in this process. In this regard, a female-biased odorant receptor of the wasp, MmedOR49, attracted our attention. OBJECTIVES: To unravel the significance of the female-biased MmedOR49 regulating host pheromone recognition. METHODS: Expression analysis (fluorescence in situ hybridization; quantitative realtime PCR), in vitro (two-electrode voltage-clamp recordings) and in vivo (RNAi combined with behavioral assessments) functional studies, and bioinformatics (structural modeling and molecular docking) were carried out to investigate the characteristics of MmedOR49. RESULTS: MmedOR49 expression was detected in the antennae of females by FISH. Quantification indicated that the expression level of MmedOR49 increased significantly after adult emergence. In vitro functional study revealed that MmedOR49 was specifically tuned to cis-5-decenyl acetate (Z5-10:Ac), the major sex pheromone component of A. segetum. Molecular docking showed that Z5-10:Ac strongly bound to the key amino acid residues His 80, Ile 81, and Arg 84 of MmedOR49 through hydrogen bonding. Behavioral assays indicated that female wasps were significantly attracted by Z5-10:Ac in a three-cage olfactometer. RNAi targeting further confirmed that MmedOR49 was necessary to recognize Z5-10:Ac, as female wasps lost their original behavioral responses to Z5-10:Ac after down-regulation of the MmedOR49 transcript. CONCLUSION: Although M. mediator is a larval endoparasitoid, female wasps have a behavioral preference for a sex pheromone component of lepidopteran hosts. In this behavior, for female M. mediator, MmedOR49 plays an important role in guiding the habitat of host insects. These data provide a potential target for enhancing natural enemy utilization and pest control.


Assuntos
Mariposas , Receptores Odorantes , Atrativos Sexuais , Vespas , Feminino , Animais , Receptores Odorantes/genética , Receptores Odorantes/metabolismo , Atrativos Sexuais/metabolismo , Hibridização in Situ Fluorescente , Simulação de Acoplamento Molecular , Vespas/genética , Vespas/metabolismo , Mariposas/genética , Mariposas/metabolismo
12.
Insect Biochem Mol Biol ; 152: 103889, 2023 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-36493964

RESUMO

The olfactory transduction system of insects is involved in multiple behavioral processes such as foraging, mating, and egg-laying behavior. In the insect olfactory receptor neurons (ORNs), the odorant receptor co-receptor (Orco) is an obligatory component that is required for dimerization with odorant receptors (ORs) to form a ligand-gated ion channel complex. The ORs/Orco heteromeric complex plays a crucial role in insect olfaction. To explore the function of OR-mediated olfaction in the physiological behavior of the fall armyworm, Spodoptera frugiperda, we applied CRISPR/Cas9 genome editing to mutate its Orco gene and constructed a homozygous mutant strain of Orco (Orco-/-) by genetic crosses. Electroantennogram (EAG) analysis showed that the responses of Orco-/- male moths to two universal sex pheromones, Z9-14: Ac and Z7-12: Ac, were abolished. We found that Orco-/- males cannot successfully mate with female moths. An oviposition preference assay confirmed that Orco-/- female moths had a reduced preference for the optimal host plant maize. A larval feeding assay revealed that the time for Orco-/- larvae to locate the food source was significantly longer than in the wild-type. Overall, in the absence of Orco, the OR-dependent olfactory behavior was impaired in both larval and adult stages. Our results confirm that Orco is essential for multiple behavioral processes related to olfaction in the fall armyworm.


Assuntos
Mariposas , Neurônios Receptores Olfatórios , Receptores Odorantes , Masculino , Feminino , Animais , Receptores Odorantes/genética , Spodoptera/genética , Oviposição , Olfato/genética , Larva/genética , Insetos
13.
Proc Natl Acad Sci U S A ; 120(1): e2214418120, 2023 Jan 03.
Artigo em Inglês | MEDLINE | ID: mdl-36584295

RESUMO

Pheromones play essential roles in reproduction in many species. Prostaglandin F2α (PGF2α) acts as a female reproductive hormone and as a sex pheromone in some species. An olfactory receptor (OR) for PGF2α was recently discovered in zebrafish, but this signaling pathway is evolutionarily labile. To understand the evolution of signals that attract males to fertile females, we used the African cichlid Astatotilapia burtoni and found that adult males strongly prefer fertile female odors. Injection of a prostaglandin synthesis inhibitor abolishes this attractivity of fertile females, indicating these hormones are necessary for pheromonal signaling. Unlike zebrafish, A. burtoni males are insensitive to PGF2α, but they do exhibit strong preference for females injected with PGF2α. This attractiveness is independent of the PGF2α hormonal receptor Ptgfr, indicating that this pheromone signaling derives from PGF2α metabolization into a yet-undiscovered pheromone. We further discovered that fish that are insensitive to PGF2α lack an ortholog for the OR Or114 that zebrafish use to detect PGF2α. These results indicate that PGF2α itself does not directly induce male preference in cichlids. Rather, it plays a vital role that primes females to become attractive via an alternative male OR.


Assuntos
Ciclídeos , Receptores Odorantes , Animais , Feminino , Masculino , Peixe-Zebra , Hormônios , Transdução de Sinais , Feromônios , Prostaglandinas
14.
Chem Senses ; 472022 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-36458901

RESUMO

Insects rely on olfactory receptors to detect and respond to diverse environmental chemical cues. Detection of semiochemicals by these receptors modulates insect behavior and has a direct impact on species fitness. Volatile organic compounds (VOCs) are released by animals and plants and can provide contextual cues that a blood meal host or nectar source is present. One such VOC is linalool, an enantiomeric monoterpene, that is emitted from plants and bacteria species. This compound exists in nature as one of two possible stereoisomers, (R)-(-)-linalool or (S)-(+)-linalool. In this study, we use a heterologous expression system to demonstrate differential responsiveness of a pair of Anopheline odorant receptors (Ors) to enantiomers of linalool. The mosquitoes Anopheles gambiae and Anopheles stephensi encode single copies of Or29 and Or53, which are expressed in the labella of An. gambiae. (S)-(+)-linalool activates Or29 orthologs with a higher potency than (R)-(-)-linalool, while the converse is observed for Or53 orthologs. The conservation of these receptors across a broad range of Anopheline species suggests they may function in the discrimination of linalool stereoisomers, thereby influencing the chemical ecology of mosquitoes. One potential application of this knowledge would be in the design of novel attractants or repellents to be used in integrated pest management practices.


Assuntos
Anopheles , Receptores Odorantes , Animais , Monoterpenos Acíclicos , Monoterpenos/farmacologia , Receptores Odorantes/genética
15.
Sci Rep ; 12(1): 22628, 2022 12 31.
Artigo em Inglês | MEDLINE | ID: mdl-36587060

RESUMO

Locusts depend upon their sense of smell and provide useful models for understanding olfaction. Extending this understanding requires knowledge of the molecular and structural organization of the olfactory system. Odor sensing begins with olfactory receptor neurons (ORNs), which express odorant receptors (ORs). In insects, ORNs are housed, in varying numbers, in olfactory sensilla. Because the organization of ORs within sensilla affects their function, it is essential to identify the ORs they contain. Here, using RNA sequencing, we identified 179 putative ORs in the transcriptomes of the two main olfactory organs, antenna and palp, of the locust Schistocerca americana. Quantitative expression analysis showed most putative ORs (140) are expressed in antennae while only 31 are in the palps. Further, our analysis identified one OR detected only in the palps and seven ORs that are expressed differentially by sex. An in situ analysis of OR expression suggested ORs are organized in non-random combinations within antennal sensilla. A phylogenetic comparison of OR predicted protein sequences revealed homologous relationships among two other Acrididae species. Our results provide a foundation for understanding the organization of the first stage of the olfactory system in S. americana, a well-studied model for olfactory processing.


Assuntos
Gafanhotos , Neurônios Receptores Olfatórios , Receptores Odorantes , Animais , Receptores Odorantes/metabolismo , Filogenia , Neurônios Receptores Olfatórios/metabolismo , Gafanhotos/genética , Gafanhotos/metabolismo , Sensilas/metabolismo , Olfato/genética , Antenas de Artrópodes/metabolismo , Proteínas de Insetos/genética , Proteínas de Insetos/metabolismo
16.
Elife ; 112022 12 13.
Artigo em Inglês | MEDLINE | ID: mdl-36511779

RESUMO

The southern house mosquito, Culex quinquefasciatus, utilizes two odorant receptors, CquiOR10 and CquiOR2, narrowly tuned to oviposition attractants and well conserved among mosquito species. They detect skatole and indole, respectively, with reciprocal specificity. We swapped the transmembrane (TM) domains of CquiOR10 and CquiOR2 and identified TM2 as a specificity determinant. With additional mutations, we showed that CquiOR10A73L behaved like CquiOR2. Conversely, CquiOR2L74A recapitulated CquiOR10 specificity. Next, we generated structural models of CquiOR10 and CquiOR10A73L using RoseTTAFold and AlphaFold and docked skatole and indole using RosettaLigand. These modeling studies suggested space-filling constraints around A73. Consistent with this hypothesis, CquiOR10 mutants with a bulkier residue (Ile, Val) were insensitive to skatole and indole, whereas CquiOR10A73G retained the specificity to skatole and showed a more robust response than the wildtype receptor CquiOR10. On the other hand, Leu to Gly mutation of the indole receptor CquiOR2 reverted the specificity to skatole. Lastly, CquiOR10A73L, CquiOR2, and CquiOR2L74I were insensitive to 3-ethylindole, whereas CquiOR2L74A and CquiOR2L74G gained activity. Additionally, CquiOR10A73G gave more robust responses to 3-ethylindole than CquiOR10. Thus, we suggest the specificity of these receptors is mediated by a single amino acid substitution, leading to finely tuned volumetric space to accommodate specific oviposition attractants.


Assuntos
Culicidae , Receptores Odorantes , Animais , Feminino , Culicidae/genética , Receptores Odorantes/genética , Receptores Odorantes/metabolismo , Escatol , Aminoácidos , Indóis , Oviposição/fisiologia
17.
BMC Genomics ; 23(1): 845, 2022 Dec 22.
Artigo em Inglês | MEDLINE | ID: mdl-36544089

RESUMO

BACKGROUND: In insects, the chemosensory system is crucial in guiding their behaviors for survival. Plagiodera versicolora (Coleoptera: Chrysomelidae), is a worldwide leaf-eating forest pest in salicaceous trees. There is little known about the chemosensory genes in P. versicolora. Here, we conducted a transcriptome analysis of larvae heads in P. versicolora. RESULTS: In this study, 29 odorant binding proteins (OBPs), 6 chemosensory proteins (CSPs), 14 odorant receptors (ORs), 13 gustatory receptors (GRs), 8 ionotropic receptors (IRs) and 4 sensory neuron membrane proteins (SNMPs) were identified by transcriptome analysis. Compared to the previous antennae and foreleg transcriptome data in adults, 12 OBPs, 2 CSPs, 5 ORs, 4 IRs, and 7 GRs were newly identified in the larvae. Phylogenetic analyses were conducted and found a new candidate CO2 receptor (PverGR18) and a new sugar receptor (PverGR23) in the tree of GRs. Subsequently, the dynamic expression profiles of various genes were analyzed by quantitative real-time PCR. The results showed that PverOBP31, OBP34, OBP35, OBP38, and OBP40 were highly expressed in larvae, PverOBP33 and OBP37 were highly expressed in pupae, and PverCSP13 was highly expressed in eggs, respectively. CONCLUSIONS: We identified a total of 74 putative chemosensory genes based on a transcriptome analysis of larvae heads in P. versicolora. This work provides new information for functional studies on the chemoreception mechanism in P. versicolora.


Assuntos
Besouros , Receptores Odorantes , Animais , Filogenia , Larva/genética , Larva/metabolismo , Perfilação da Expressão Gênica , Transcriptoma , Besouros/genética , Receptores Odorantes/genética , Receptores Odorantes/metabolismo , Proteínas de Insetos/genética , Proteínas de Insetos/metabolismo , Antenas de Artrópodes/metabolismo
18.
Cells ; 11(24)2022 12 07.
Artigo em Inglês | MEDLINE | ID: mdl-36552724

RESUMO

In recent years, there has been a great deal of interest in the ectopic roles of olfactory receptors (ORs) throughout the human body. Especially, the ectopic function of OR in the skin is one of the most actively researched areas. Suberic acid, a scent compound, was hypothesized to increase collagen synthesis in the ultraviolet B (UVB)-irradiated human dermal fibroblasts (Hs68) through a specific olfactory receptor. Suberic acid ameliorated UVB-induced decreases in collagen production in Hs68 cells. Using in silico docking to predict the binding conformation and affinity of suberic acid to 15 ectopic ORs detectable in Hs68, several ORs were identified as promising candidates. The effect of suberic acid on collagen synthesis in UVB-exposed dermal fibroblasts was nullified only by a reduction in OR10A3 expression via specific siRNA. In addition, using the cells transiently expressing OR10A3, we demonstrated that suberic acid can activate OR10A3 by assessing the downstream effector cAMP response element (CRE) luciferase activity. We examined that the activation of OR10A3 by suberic acid subsequently stimulates collagen synthesis via the downstream cAMP-Akt pathway. The findings support OR10A3 as a promising target for anti-aging treatments of the skin.


Assuntos
Receptores Odorantes , Envelhecimento da Pele , Humanos , Colágeno/metabolismo , Fibroblastos/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , Envelhecimento da Pele/efeitos dos fármacos , Envelhecimento da Pele/genética , Envelhecimento da Pele/fisiologia , Receptores Odorantes/genética , Receptores Odorantes/metabolismo
19.
Mol Med ; 28(1): 150, 2022 Dec 12.
Artigo em Inglês | MEDLINE | ID: mdl-36503361

RESUMO

BACKGROUND: Therapeutic options for steroid-resistant non-type 2 inflammation in obstructive lung diseases are lacking. Alveolar macrophages are central in the progression of these diseases by releasing proinflammatory cytokines, making them promising targets for new therapeutic approaches. Extra nasal expressed olfactory receptors (ORs) mediate various cellular processes, but clinical data are lacking. This work investigates whether ORs in human primary alveolar macrophages could impact pathophysiological processes and could be considered as therapeutic targets. METHODS: Human primary alveolar macrophages were isolated from bronchoalveolar lavages of 50 patients with pulmonary diseases. The expression of ORs was validated using RT-PCR, immunocytochemical staining, and Western blot. Changes in intracellular calcium levels were analyzed in real-time by calcium imaging. A luminescent assay was used to measure the cAMP concentration after OR stimulation. Cytokine secretion was measured in cell supernatants 24 h after stimulation by ELISA. Phagocytic ability was measured by the uptake of fluorescent-labeled beads by flow cytometry. RESULTS: We demonstrated the expression of functional OR2AT4 and OR1A2 on mRNA and protein levels. Both ORs were primarily located in the plasma membrane. Stimulation with Sandalore, the ligand of OR2AT4, and Citronellal, the ligand of OR1A2, triggered a transient increase of intracellular calcium and cAMP. In the case of Sandalore, this calcium increase was based on a cAMP-dependent signaling pathway. Stimulation of alveolar macrophages with Sandalore and Citronellal reduced phagocytic capacity and release of proinflammatory cytokines. CONCLUSION: These are the first indications for utilizing olfactory receptors as therapeutic target molecules in treating steroid-resistant lung diseases with non-type 2 inflammation.


Assuntos
Pneumopatias , Receptores Odorantes , Humanos , Cálcio/metabolismo , Citocinas/metabolismo , Inflamação/metabolismo , Ligantes , Pneumopatias/metabolismo , Macrófagos Alveolares/metabolismo , Receptores Odorantes/genética , Receptores Odorantes/metabolismo , Esteroides
20.
Int J Mol Sci ; 23(24)2022 Dec 12.
Artigo em Inglês | MEDLINE | ID: mdl-36555416

RESUMO

Scopula subpunctaria, an abundant pest in tea gardens, produce type-II sex pheromone components, which are critical for its communicative and reproductive abilities; however, genes encoding the proteins involved in the detection of type-II sex pheromone components have rarely been documented in moths. In the present study, we sequenced the transcriptomes of the male and female S. subpunctaria antennae. A total of 150 candidate olfaction genes, comprising 58 odorant receptors (SsubORs), 26 ionotropic receptors (SsubIRs), 24 chemosensory proteins (SsubCSPs), 40 odorant-binding proteins (SsubOBPs), and 2 sensory neuron membrane proteins (SsubSNMPs) were identified in S. subpunctaria. Phylogenetic analysis, qPCR, and mRNA abundance analysis results suggested that SsubOR46 may be the Orco (non-traditional odorant receptor, a subfamily of ORs) of S. subpunctaria. SsubOR9, SsubOR53, and SsubOR55 belonged to the pheromone receptor (PR) clades which have a higher expression in male antennae. Interestingly, SsubOR44 was uniquely expressed in the antennae, with a higher expression in males than in females. SsubOBP25, SsubOBP27, and SsubOBP28 were clustered into the moth pheromone-binding protein (PBP) sub-family, and they were uniquely expressed in the antennae, with a higher expression in males than in females. SsubOBP19, a member of the GOBP2 group, was the most abundant OBP in the antennae. These findings indicate that these olfactory genes, comprising five candidate PRs, three candidate PBPs, and one candidate GOBP2, may be involved in type II sex pheromone detection. As well as these genes, most of the remaining SsubORs, and all of the SsubIRs, showed a considerably higher expression in the female antennae than in the male antennae. Many of these, including SsubOR40, SsubOR42, SsubOR43, and SsubIR26, were more abundant in female antennae. These olfactory and ionotropic receptors may be related to the detection of host plant volatiles. The results of this present study provide a basis for exploring the olfaction mechanisms in S. subpunctaria, with a focus on the genes involved in type II sex pheromones. The evolutionary analyses in our study provide new insights into the differentiation and evolution of lepidopteran PRs.


Assuntos
Mariposas , Receptores Odorantes , Atrativos Sexuais , Animais , Feminino , Masculino , Atrativos Sexuais/genética , Atrativos Sexuais/metabolismo , Filogenia , Olfato/genética , Perfilação da Expressão Gênica/métodos , Mariposas/genética , Mariposas/metabolismo , Receptores Odorantes/genética , Receptores Odorantes/metabolismo , Proteínas de Insetos/genética , Proteínas de Insetos/metabolismo , Antenas de Artrópodes/metabolismo
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