RESUMO
In many arthropods, including insects responsible for transmission of human diseases, behaviors that include mating, aggregation, and aggression are triggered by detection of pheromones. Extracellular odorant binding proteins are critical for pheromone detection in many insects and are secreted into the fluid bathing the olfactory neuron dendrites. In Drosophila melanogaster, the odorant binding protein LUSH is essential for normal sensitivity to the volatile sex pheromone, 11-cis vaccenyl acetate (cVA). Using a genetic screen for cVA pheromone insensitivity, we identified ANCE-3, a homolog of human angiotensin converting enzyme that is required for detection of cVA pheromone. The mutants have normal dose-response curves for food odors, although olfactory neuron amplitudes are reduced in all olfactory neurons examined. ance-3 mutants have profound delays in mating, and the courtship defects are primarily but not exclusively due to loss of ance-3 function in males. We demonstrate that ANCE-3 is required in the sensillae support cells for normal reproductive behavior, and that localization of odorant binding proteins to the sensillum lymph is blocked in the mutants. Expression of an ance-3 cDNA in sensillae support cells completely rescues the cVA responses, LUSH localization, and courtship defects. We show the courtship latency defects are not due to effects on olfactory neurons in the antenna nor mediated through ORCO receptors, but instead stem from ANCE-3-dependent effects on chemosensory sensillae in other body parts. These findings reveal an unexpected factor critical for pheromone detection with profound influence on reproductive behaviors.
Assuntos
Proteínas de Drosophila , Receptores Odorantes , Animais , Humanos , Masculino , Corte , Drosophila melanogaster/metabolismo , Proteínas de Drosophila/genética , Proteínas de Drosophila/metabolismo , Odorantes , Peptidil Dipeptidase A , Feromônios/metabolismo , Receptores Odorantes/genética , Receptores Odorantes/metabolismo , Comportamento Sexual Animal/fisiologiaRESUMO
Sex pheromones are pivotal for insect reproduction. However, the mechanism of sex pheromone communication remains enigmatic in hymenopteran parasitoids. Here we have identified the sex pheromone and elucidated the olfactory basis of sex pheromone communication in Campoletis chlorideae (Ichneumonidae), a solitary larval endoparasitoid of over 30 lepidopteran pests. Using coupled gas chromatography-electroantennogram detection, we identified two female-derived pheromone components, tetradecanal (14:Ald) and 2-heptadecanone (2-Hep) (1:4.6), eliciting strong antennal responses from males but weak responses from females. We observed that males but not females were attracted to both single components and the blend. The hexane-washed female cadavers failed to arouse males, and replenishing 14:Ald and 2-Hep could partially restore the sexual attraction of males. We further expressed six C. chlorideae male-biased odorant receptors in Drosophila T1 neurons and found that CchlOR18 and CchlOR47 were selectively tuned to 14:Ald and 2-Hep, respectively. To verify the biological significance of this data, we knocked down CchlOR18 and CchlOR47 individually or together in vivo and show that the attraction of C. chlorideae to their respective ligands was abolished. Moreover, the parasitoids defective in either of the receptors were less likely to court and copulate. Finally, we show that the sex pheromone and (Z)-jasmone, a potent female attractant, can synergistically affect behaviors of virgin males and virgin females and ultimately increase the parasitic efficiency of C. chlorideae. Our study provides new insights into the molecular mechanism of sex pheromone communication in C. chlorideae that may permit manipulation of parasitoid behavior for pest control.
Assuntos
Receptores Odorantes , Atrativos Sexuais , Masculino , Animais , Insetos , Comunicação , Feromônios , DrosophilaRESUMO
Human and insect olfaction share many general features, but insects differ from mammalian systems in important ways. Mammalian olfactory neurons share the same overlying fluid layer in the nose, and neuronal tuning entirely depends upon receptor specificity. In insects, the olfactory neurons are anatomically segregated into sensilla, and small clusters of olfactory neurons dendrites share extracellular fluid that can be independently regulated in different sensilla. Small extracellular proteins called odorant-binding proteins are differentially secreted into this sensillum lymph fluid where they have been shown to confer sensitivity to specific odorants, and they can also affect the kinetics of the olfactory neuron responses. Insect olfactory receptors are not G-protein-coupled receptors, such as vertebrate olfactory receptors, but are ligand-gated ion channels opened by direct interactions with odorant molecules. Recently, several examples of insect olfactory neurons expressing multiple receptors have been identified, indicating that the mechanisms for neuronal tuning may be broader in insects than mammals. Finally, recent advances in genome editing are finding applications in many species, including agricultural pests and human disease vectors.
RESUMO
We evaluated whether intraverbal and reverse intraverbal behavior emerged following listener training in children with autism spectrum disorder (ASD). Six participants were each taught three sets of three "when?" questions in listener training. A multiple baseline design across behaviors (stimulus sets) was used to assess the effects of listener training. Results showed that intraverbal behavior emerged following listener training for five out of six participants. One participant received additional listener training and intraverbal training before intraverbal behavior emerged. Furthermore, reverse intraverbal responding occurred across all three sets of questions for three of the six participants. Establishing listener behavior may be a pathway for emergent intraverbal and reverse intraverbal responding in children with ASD. Future research could examine what skill repertoire may facilitate such transfer.
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Insects use olfaction to detect ecologically relevant chemicals in their environment. To maintain useful responses over a variety of stimuli, olfactory receptor neurons are desensitized to prolonged or high concentrations of stimuli. Depending on the timescale, the desensitization is classified as short-term, which typically spans a few seconds; or long-term, which spans from minutes to hours. Compared with the well-studied mechanisms of desensitization in vertebrate olfactory neurons, the mechanisms underlying invertebrate olfactory sensitivity regulation remain poorly understood. Recently, using a large-scale functional screen, a conserved critical receptor phosphorylation site has been identified in the model insect Drosophila melanogaster, providing new insight into the molecular basis of desensitization in insects. Here, we summarize the progress in this area and provide perspectives on future directions to determine the molecular mechanisms that orchestrate the desensitization in insect olfaction.
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Alcohol-induced aggression is a destructive and widespread phenomenon associated with violence and sexual assault. However, little is understood concerning its mechanistic origin. We have developed a Drosophila melanogaster model to genetically dissect and understand the phenomenon of sexually dimorphic alcohol-induced aggression. Males with blood alcohol levels of 0.04-mg/ml BAC were less aggressive than alcohol-naive males, but when the BAC had dropped to ~0.015 mg/ml, the alcohol-treated males showed an increase in aggression toward other males. This aggression-promoting treatment is referred to as the post-ethanol aggression (PEA) treatment. Females do not show increased aggression after the same treatment. PEA-treated males also spend less time courting and attempt to copulate earlier than alcohol-naive flies. PEA treatment induces expression of the FruM transcription factor (encoded by a male-specific transcript from the fruitless gene), whereas sedating doses of alcohol reduce FruM expression and reduce male aggression. Transgenic suppression of FruM induction also prevents alcohol-induced aggression. In male flies, alcohol-induced aggression is dependent on the male isoform of the fruitless transcription factor (FruM). Low-dose alcohol induces FruM expression and promotes aggression, whereas higher doses of alcohol suppress FruM and suppress aggression.
Assuntos
Agressão , Etanol/metabolismo , Comportamento Sexual Animal/efeitos dos fármacos , Animais , Drosophila melanogaster , Feminino , Regulação da Expressão Gênica , Masculino , Proteínas do Tecido Nervoso/metabolismo , Neurônios/metabolismo , Caracteres Sexuais , Fatores de TranscriçãoRESUMO
Drosophila odorant receptors (Ors) are ligand gated ion channels composed of a common receptor subunit Or co-receptor (ORCO) and one of 62 "tuning" receptor subunits that confer odorant specificity to olfactory neuron responses. Like other sensory systems studied to date, exposing Drosophila olfactory neurons to activating ligands results in reduced responses to subsequent exposures through a process called desensitization. We recently showed that phosphorylation of serine 289 on the common Or subunit ORCO is required for normal peak olfactory neuron responses. Dephosphorylation of this residue occurs on prolonged odorant exposure, and underlies the slow modulation of olfactory neuron responses we term "slow desensitization." Slow desensitization results in the reduction of peak olfactory neuron responses and flattening of dose-response curves, implicating changes in ORCOS289 phosphorylation state as an important modulator of olfactory neuron responses. Here, we report the identification of the primary kinase responsible for ORCOS289 phosphorylation, PKC98E. Antiserum localizes the kinase to the dendrites of the olfactory neurons. Deletion of the kinase from olfactory neurons in the naive state (the absence of prolonged odor exposure) reduces ORCOS289 phosphorylation and reduces peak odorant responses without altering receptor localization or expression levels. Genetic rescue with a PKC98E predicted to be constitutively active restores ORCO S289 phosphorylation and olfactory neuron sensitivity to the PKC98E mutants in the naive state. However, the dominant kinase is defective for slow desensitization. Together, these findings reveal that PKC98E is an important regulator of ORCO receptors and olfactory neuron function.SIGNIFICANCE STATEMENT We have identified PKC98E as the kinase responsible for phosphorylation of the odorant receptor co-receptor (ORCO) at S289 that is required for normal odorant response kinetics of olfactory neurons. This is a significant step toward revealing the enzymology underlying the regulation of odorant response regulation in insects.
Assuntos
Proteínas de Drosophila/fisiologia , Drosophila melanogaster/fisiologia , Neurônios Receptores Olfatórios/fisiologia , Proteína Quinase C/fisiologia , Animais , Dendritos/enzimologia , Relação Dose-Resposta a Droga , Proteínas de Drosophila/genética , Fenômenos Eletrofisiológicos , Deleção de Genes , Mutação/genética , Odorantes , Neurônios Receptores Olfatórios/enzimologia , Fosforilação , Proteína Quinase C/genética , Interferência de RNA , Receptores Odorantes/genética , Receptores Odorantes/metabolismoRESUMO
This study reports outcome in adolescents with autism who in their childhood received Early and Intensive Behavioral Intervention (EIBI). Nineteen children (16 boys) who had received two years of EIBI starting at a mean age of 2-years-and-11-months were followed up, on average, 12 years later. Results showed the participants significantly increased their cognitive and adaptive standard scores during the two years of EIBI, and that these gains were maintained at follow-up, 10 years after the EIBI had ended. Participants also showed a significant reduction in autism symptoms between intake and follow-up. At follow-up, none of the participants had received any additional psychiatric diagnoses, and none were taking any psychotropic medication. Results indicate that treatment gains achieved in EIBI are maintained into adolescence.
Assuntos
Transtorno Autístico , Intervenção Médica Precoce , Adolescente , Transtorno Autístico/terapia , Terapia Comportamental , Criança , Pré-Escolar , Intervenção Educacional Precoce , Humanos , MasculinoRESUMO
For decades, numerous researchers have documented the presence of the fruit fly or Drosophila melanogaster on alcohol-containing food sources. Although fruit flies are a common laboratory model organism of choice, there is relatively little understood about the ethological relationship between flies and ethanol. In this study, we find that when male flies inhabit ethanol-containing food substrates they become more aggressive. We identify a possible mechanism for this behavior. The odor of ethanol potentiates the activity of sensory neurons in response to an aggression-promoting pheromone. Finally, we observed that the odor of ethanol also promotes attraction to a food-related citrus odor. Understanding how flies interact with the complex natural environment they inhabit can provide valuable insight into how different natural stimuli are integrated to promote fundamental behaviors.
Assuntos
Drosophila melanogaster/fisiologia , Etanol/metabolismo , Feromônios/metabolismo , Agressão , Animais , Comportamento Animal , Feminino , Masculino , Odorantes/análiseRESUMO
Insect odorant-binding proteins (OBPs) are a large, diverse group of low-molecular weight proteins secreted into the fluid bathing olfactory and gustatory neuron dendrites. The best-characterized OBP, LUSH (OBP76a) enhances pheromone sensitivity enabling detection of physiological levels of the male-specific pheromone, 11-cis vaccenyl acetate. The role of the other OBPs encoded in the Drosophila genome is largely unknown. Here, using clustered regularly interspaced short palindromic repeats/Cas9, we generated and characterized the loss-of-function phenotype for two genes encoding homologous OBPs, OS-E (OBP83b) and OS-F (OBP83a). Instead of activation defects, these extracellular proteins are required for normal deactivation of odorant responses to a subset of odorants. Remarkably, odorants detected by the same odorant receptor are differentially affected by the loss of the OBPs, revealing an odorant-specific role in deactivation kinetics. In stark contrast to lush mutants, the OS-E/F mutants have normal activation kinetics to the affected odorants, even at low stimulus concentrations, suggesting that these OBPs are not competing for these ligands with the odorant receptors. We also show that OS-E and OS-F are functionally redundant as either is sufficient to revert the mutant phenotype in transgenic rescue experiments. These findings expand our understanding of the roles of OBPs to include the deactivation of odorant responses.
Assuntos
Proteínas de Transporte/genética , Proteínas de Drosophila/genética , Mutação com Perda de Função , Fenótipo , Animais , Proteínas de Transporte/metabolismo , Proteínas de Drosophila/metabolismo , Drosophila melanogaster , Ligação ProteicaRESUMO
Insects and other arthropods transmit devastating human diseases, and these vectors use chemical senses to target humans. Understanding how these animals detect, respond, and adapt to volatile odorants may lead to novel ways to disrupt host localization or mate recognition in these pests. The past decade has led to remarkable progress in understanding odorant detection in arthropods. Insects use odorant-gated ion channels, first discovered in Drosophila melanogaster, to detect volatile chemicals. In flies, 60 "tuning" receptor subunits combine with a common subunit, Orco (odorant receptor coreceptor) to form ligand-gated ion channels. The mechanisms underlying odorant receptor desensitization in insects are largely unknown. Recent work reveals that dephosphorylation of serine 289 on the shared Orco subunit is responsible for slow, odor-induced receptor desensitization. Dephosphorylation has no effect on the localization of the receptor protein, and activation of the olfactory neurons in the absence of odor is sufficient to induce dephosphorylation and desensitization. These findings reveal a major component of receptor modulation in this important group of disease vectors, and implicate a second messenger feedback mechanism in this process.
RESUMO
Insects have evolved sophisticated olfactory reception systems to sense exogenous chemical signals. Odorant receptors (ORs) on the membrane of chemosensory neurons are believed to be key molecules in sensing exogenous chemical cues. ORs in different species of insects are diverse and should tune a species to its own specific semiochemicals relevant to their survival. The orthopteran insect, locust (Locusta migratoria), is a model hemimetabolous insect. There is very limited knowledge on the functions of locust ORs although many locust OR genes have been identified in genomic sequencing experiments. In this paper, a locust OR, LmigOR3 was localized to neurons housed in trichoid sensilla by in situ hybridization. LmigOR3 was expressed as a transgene in Drosophila trichoid olfactory neurons (aT1) lacking the endogenous receptor Or67d and the olfactory tuning curve and dose-response curves were established for this locust receptor. The results show that LmigOR3 sensitizes neurons to ketones, esters and heterocyclic compounds, indicating that LmigOR3 is a broadly tuned receptor. LmigOR3 is the first odorant receptor from Orthoptera that has been functionally analyzed in the Drosophila aT1 system. This work demonstrates the utility of the Drosophila aT1 system for functional analysis of locust odorant receptors and suggests that LmigOR3 may be involved in detecting food odorants, or perhaps locust body volatiles that may help us to develop new control methods for locusts.
Assuntos
Locusta migratoria/genética , Receptores Odorantes/genética , Animais , Animais Geneticamente Modificados/genética , Animais Geneticamente Modificados/metabolismo , Drosophila melanogaster/genética , Drosophila melanogaster/metabolismo , Locusta migratoria/metabolismo , Neurônios/metabolismo , Receptores Odorantes/metabolismo , Sensilas/metabolismoRESUMO
The purpose of the present study was to assess whether intraverbal behavior, in the form of answers to questions, emerges as a result of listener training for five children diagnosed with autism. Listener responses were targeted and taught using prompting and differential reinforcement. Following successful acquisition of listener responses, the intraverbal form of the response was probed. Data were evaluated via a nonconcurrent multiple-baseline design that included a control series. Results showed listener-to-intraverbal transfer for four of the five participants. One participant required additional teaching that involved tacting the items selected during listener training.
RESUMO
In Drosophila melanogaster, the male-specific pheromone cVA (11-cis-vaccenyl acetate) functions as a sex-specific social cue. However, our understanding of the molecular mechanisms underlying cVA pheromone transduction and its regulation are incomplete. Using a genetic screen combined with an electrophysiological assay to monitor pheromone-evoked activity in the cVA-sensing Or67d neurons, we identified an olfactory sensitivity factor encoded by the dATP8B gene, the Drosophila homolog of mammalian ATP8B. dATP8B is expressed in all olfactory neurons that express Orco, the odorant receptor coreceptor, and the odorant responses in most Orco-expressing neurons are reduced. Or67d neurons are severely affected, with strongly impaired cVA-induced responses and lacking spontaneous spiking in the mutants. The dATP8B locus encodes a member of the P4-type ATPase family thought to flip aminophospholipids such as phosphatidylserine and phosphatidylethanolamine from one membrane leaflet to the other. dATP8B protein is concentrated in the cilia of olfactory neuron dendrites, the site of odorant transduction. Focusing on Or67d neuron function, we show that Or67d receptors are mislocalized in dATP8B mutants and that cVA responses can be restored to dATP8B mutants by misexpressing a wild-type dATP8B rescuing transgene, by expressing a vertebrate P4-type ATPase member in the pheromone-sensing neurons or by overexpressing Or67d receptor subunits. These findings reveal an unexpected role for lipid translocation in olfactory receptor expression and sensitivity to volatile odorants.
Assuntos
Proteínas de Drosophila/metabolismo , Drosophila melanogaster/fisiologia , Ácidos Oleicos/metabolismo , Feromônios/metabolismo , Proteínas de Transferência de Fosfolipídeos/metabolismo , Receptores de Superfície Celular/metabolismo , Receptores Odorantes/metabolismo , Olfato/fisiologia , Animais , Sequência de Bases , Primers do DNA/genética , Proteínas de Drosophila/genética , Processamento de Imagem Assistida por Computador , Imuno-Histoquímica , Dados de Sequência Molecular , Proteínas de Transferência de Fosfolipídeos/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Análise de Sequência de DNARESUMO
We set out to deorphanize a subset of putative Drosophila odorant receptors expressed in trichoid sensilla using a transgenic in vivo misexpression approach. We identified farnesol as a potent and specific activator for the orphan odorant receptor Or83c. Farnesol is an intermediate in juvenile hormone biosynthesis, but is also produced by ripe citrus fruit peels. Here, we show that farnesol stimulates robust activation of Or83c-expressing olfactory neurons, even at high dilutions. The CD36 homolog Snmp1 is required for normal farnesol response kinetics. The neurons expressing Or83c are found in a subset of poorly characterized intermediate sensilla. We show that these neurons mediate attraction behavior to low concentrations of farnesol and that Or83c receptor mutants are defective for this behavior. Or83c neurons innervate the DC3 glomerulus in the antennal lobe and projection neurons relaying information from this glomerulus to higher brain centers target a region of the lateral horn previously implicated in pheromone perception. Our findings identify a sensitive, narrowly tuned receptor that mediates attraction behavior to farnesol and demonstrates an effective approach to deorphanizing odorant receptors expressed in neurons located in intermediate and trichoid sensilla that may not function in the classical "empty basiconic neuron" system.
Assuntos
Proteínas de Drosophila/fisiologia , Drosophila melanogaster/fisiologia , Farneseno Álcool , Neurônios Receptores Olfatórios/fisiologia , Receptores Odorantes/fisiologia , Animais , Animais Geneticamente Modificados , Comportamento Animal/fisiologia , Proteínas de Drosophila/genética , Drosophila melanogaster/anatomia & histologia , Feminino , Gânglios dos Invertebrados/citologia , Gânglios dos Invertebrados/fisiologia , Masculino , Proteínas de Membrana/fisiologia , Proteínas do Tecido Nervoso/fisiologia , Odorantes , Receptores Odorantes/genética , Olfato/fisiologiaRESUMO
BACKGROUND: Sensory neuron diversity ensures optimal detection of the external world and is a hallmark of sensory systems. An extreme example is the olfactory system, as individual olfactory receptor neurons (ORNs) adopt unique sensory identities by typically expressing a single receptor gene from a large genomic repertoire. In Drosophila, about 50 different ORN classes are generated from a field of precursor cells, giving rise to spatially restricted and distinct clusters of ORNs on the olfactory appendages. Developmental strategies spawning ORN diversity from an initially homogeneous population of precursors are largely unknown. RESULTS: Here we unravel the nested and binary logic of the combinatorial code that patterns the decision landscape of precursor states underlying ORN diversity in the Drosophila olfactory system. The transcription factor Rotund (Rn) is a critical component of this code that is expressed in a subset of ORN precursors. Addition of Rn to preexisting transcription factors that assign zonal identities to precursors on the antenna subdivides each zone and almost exponentially increases ORN diversity by branching off novel precursor fates from default ones within each zone. In rn mutants, rn-positive ORN classes are converted to rn-negative ones in a zone-specific manner. CONCLUSIONS: We provide a model describing how nested and binary changes in combinations of transcription factors could coordinate and pattern a large number of distinct precursor identities within a population to modulate the level of ORN diversity during development and evolution.
Assuntos
Proteínas de Drosophila/fisiologia , Drosophila/fisiologia , Receptores Odorantes/fisiologia , Olfato/fisiologia , Animais , Antenas de Artrópodes/citologia , Antenas de Artrópodes/metabolismo , Antenas de Artrópodes/fisiologia , Drosophila/citologia , Drosophila/genética , Proteínas de Drosophila/genética , Proteínas de Drosophila/metabolismo , Modelos Biológicos , Receptores Odorantes/metabolismo , Olfato/genética , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo , Fatores de Transcrição/fisiologiaRESUMO
Sweet and bitter taste distinguishes good food sources from potential toxins, but what happens when these tastants are mixed? In this issue of Neuron, Jeong et al. (2013) show that in Drosophila, bitter compounds act through an extracellular odorant-binding protein to inhibit sweet-responsive neurons and block the response to sweet taste.
Assuntos
Odorantes , Receptores Odorantes/genética , Receptores Odorantes/metabolismo , Paladar/efeitos dos fármacos , Paladar/genética , AnimaisRESUMO
A common characteristic of the language deficits experienced by children with autism (and other developmental disorders) is their failure to acquire a complex intraverbal repertoire. The difficulties with learning intraverbal behaviors may, in part, be related to the fact that the stimulus control for such behaviors usually involves highly complex verbal stimuli. The antecedent verbal control of intraverbal behavior may involve discriminative stimuli (i.e., discriminated operants), conditional stimulus control, and/or control by compound stimuli. Distinctions among these different types of antecedent control are presented, along with recommendations for intervention procedures that may facilitate the acquisition of intraverbal behavior.
RESUMO
Once captured by the antenna, 11-cis vaccenyl acetate (cVA) binds to an extracellular binding protein called LUSH that undergoes a conformational shift upon cVA binding. The stable LUSH-cVA complex is the activating ligand for pheromone receptors present on the dendrites of the aT1 neurones, comprising the only neurones that detect cVA pheromone. This mechanism explains the single molecule sensitivity of insect pheromone detection systems. The receptor that recognizes activated LUSH consists of a complex of several proteins, including Or67d, a member of the tuning odourant receptor family, Orco, a co-receptor ion channel, and SNMP, a CD36 homologue that may be an inhibitory subunit. In addition, genetic screens and reconstitution experiments reveal additional factors that are important for pheromone detection. Identification and functional dissection of these factors in Drosophila melanogaster Meigen should permit the identification of homologous factors in pathogenic insects and agricultural pests, which, in turn, may be viable candidates for novel classes of compounds to control populations of target insect species without impacting beneficial species.
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In two experiments, the authors evaluated the extent to which (a) individuals preferred engaging in object stereotypy versus observing an experimenter while the experimenter engaged in object stereotypy and (b) an experimenter's engagement in object stereotypy decreased the participants' engagement in object stereotypy. Results of Experiment 1 indicated that behaver-controlled (BC) object stereotypy was preferred over experimenter-controlled (EC) object stereotypy by three of four participants. Results of Experiment 2 indicated that EC object stereotypy decreased object stereotypy for only one of three participants. Implications of these findings for determining the relative importance of control over stimulation generated by stereotypy are briefly discussed.