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The biosynthesis and termination of insect sex pheromones should be accurately regulated. In most moths, the biosynthesis and release of sex pheromones are regulated by a class of neuropeptides known as pheromone biosynthesis activating neuropeptides (PBANs). However, endogenous mechanisms underlying the termination of sex pheromone biosynthesis in moths remain elusive. In the present study, Helicoverpa armigera was employed as a model to investigate the role of octopamine (OA) in the inhibition of sex pheromone biosynthesis. Results demonstrated that the release of sex pheromones decreased with an increase in OA titres in older female moths. Moreover, OA treatment led to a significant decrease in sex pheromone production, female capability to attract male counterparts and subsequent female acceptance, indicating its inhibitory role in sex pheromone release. Subsequent qPCR and RNAi analyses revealed that OctβR was a key receptor of OA that regulated sex pheromone biosynthesis. In addition, the OA/OctβR signal suppressed intracellular Ca2+ levels and attenuated PBAN‐mediated increase in the enzyme activities of calcineurin and acetyl‐CoA carboxylase as demonstrated by OA treatment and OctβR‐RNAi. Altogether, these results revealed a mechanism underlying the inhibition of sex pheromone production by OA via suppression of PBAN signalling in moths. [ABSTRACT FROM AUTHOR]

Insect octopamine (OA) receptors are G‐protein coupled receptors (GPCRs) that play essential roles in physiological and behavioural processes. However, there is little information about the function of OA receptors in the aphids' response to stress. From the genome sequence of Rhopalosiphum padi genome sequence, a cosmopolitan cereal pest, we identified six OA receptor genes RpOAMB, RpOctR, RpOctβ1R, RpOctβ2R, RpOctβ3R, RpOctR‐like with two, one, one, four, four, seven exons, respectively. All the OA receptors contain seven transmembrane domains, which were the signature of GPCRs. Our results showed that (1) the contents of OA increased significantly after food starvation, (2) the transcription levels of RpOAMB, RpOctR, RpOctβ2R and RpOctβ3R increased after starvation and were restored after re‐feeding, and (3) the expression levels of these four genes decreased significantly 48 h post‐injection of dsRNA that targeted the respective genes. Knockdown of RpOctR, RpOctβ2R or RpOctβ3R genes significantly increased aphid mortality under 24 h starvation conditions. Mortality of R. padi injected with dsRpOctR or dsRpOctβ2R was significantly higher than control under 48 h starvation treatments. This is the first report on the role of OA receptors in the starvation response of aphids. The current study provides knowledge for a better understanding the physiological roles of insect OA receptors. [ABSTRACT FROM AUTHOR]

The brown planthopper (BPH), Nilaparvata lugens (Stål) is a resurgent pest of rice crops throughout Asia. We recently discovered that octopamine (OA) and OA2B2 operate in the BPH mating system, where it mediates a wide range of molecular, physiological and behavioural changes. Here, we report on outcomes of experiments designed to test the hypothesis that OA/OA2B2 signalling mediates responses to three abiotic stressors, starvation, high temperature (37 °C), and induced oxidative stress. We found per os RNAi‐mediated OA2B2 silencing led to significantly decreased survival, measured in days, following exposure to each of these stressors. We selected a biologically costly process, reproductive biology, as a biotic stressor. Silencing of OA2B2 led to decreased total protein content in ovaries and fat bodies, downregulated expression of vitellogenin (Vg) and Vg receptor (VgR), inhibited fat body Vg protein synthesis, shortened the oviposition period, prolonged the preoviposition period, reduced the number of laid eggs, body weight and female longevity. In addition, the silencing treatments also led to inhibited ovarian development, and ovarian Vg uptake, reduced numbers of egg masses and offspring and lower hatching rates and population growth index. These data support our hypothesis that OA2B2 acts in mediating BPH resistance to biotic and abiotic stressors. [ABSTRACT FROM AUTHOR]

The migratory locust ( Locusta migratoria) shows aggregative traits in nymph marching bands and swarm formations through mutual olfactory attraction of conspecifics. However, olfactory preference in different nymph stages in gregarious locusts is not sufficiently explored. In this study, we found that the nymph olfactory preference for gregarious volatiles exhibited obvious variations at different developmental stages. The gregarious locusts show attractive response to conspecific volatiles from the third stadium. Transcriptome comparison between third- and fourth-stadium nymphs showed that the G protein-coupled receptor (GPCR) pathways are significantly enriched. Amongst the genes present in GPCR pathways, the expression level of calmodulin in locust brains significantly increased from the third- to the fourth-stadium nymphs. Amongst the four octopamine receptors (OARs) belonging to the GPCR family, only OAR α1 showed similar expression patterns to those of calmodulin, and knockdown of OAR α1 reduced the expression level of calmodulin. RNA interference of calmodulin decreased locomotion and induced the loss of olfactory attraction in gregarious locusts. Moreover, the activation of OAR α1 in calmodulin-knockdown locusts did not induce olfactory attraction of the nymphs to gregarious volatiles. Thus, calmodulin as a downstream gene of octopamine-OAR α1 (OA-OAR α1) signalling mediates olfactory attraction in gregarious locusts. Overall, this study provides novel insights into the mechanism of OA-OAR α1 signalling involved in olfactory attraction of gregarious locusts. [ABSTRACT FROM AUTHOR]

Nosema ceranae, an emerging pathogen of the western honeybee ( Apis mellifera), is implicated in recent pollinator losses and causes severe energetic stress. However, whether precocious foraging and accelerated behavioural maturation in infected bees are caused by the infection itself or via indirect energetic stress remains unknown. Using a combination of nutritional and infection treatments, we investigated how starvation and infection alters the regulation of adipokinetic hormone (AKH) and octopamine, two highly conserved physiological pathways that respond to energetic stress by mobilizing fat stores and increasing search activity for food. Although there was no response from AKH when bees were experimentally infected with N. ceranae or starved, supporting the notion that honeybees have lost this pathway, there were significant regulatory changes in the octopamine pathway. Significantly, we found no evidence of acute energetic stress being the only cause of symptoms associated with N. ceranae infection. Therefore, the parasite itself appears to alter regulatory components along a highly conserved physiological pathway in an infection-specific manner. This indicates that pathogen-induced behavioural alteration of chronically infected bees should not just be viewed as a coincidental short-term by-product of pathogenesis (acute energetic stress) and may be a result of a generalist manipulation strategy to obtain energy for reproduction. [ABSTRACT FROM AUTHOR]

The honey bee ( Apis mellifera L.) has developed into an important ethological model organism for social behaviour and behavioural plasticity. Bees perform a complex age-dependent division of labour with the most pronounced behavioural differences occurring between in-hive bees and foragers. Whereas nurse bees, for example, stay inside the hive and provide the larvae with food, foragers leave the hive to collect pollen and nectar for the entire colony. The biogenic amine octopamine appears to play a major role in division of labour but the molecular mechanisms involved are unknown. We here investigated the role of two characterized octopamine receptors in honey bee division of labour. Am Octα R1 codes for a Ca2+-linked octopamine receptor. Am Octβ R3/4 codes for a cyclic adenosine monophosphate-coupled octopamine receptor. Messenger RNA expression of Am Octα R1 in different brain neuropils correlates with social task, whereas expression of Am Octβ R3/4 changes with age rather than with social role per se. Our results for the first time link the regulatory role of octopamine in division of labour to specific receptors and brain regions. They are an important step forward in our understanding of complex behavioural organization in social groups. [ABSTRACT FROM AUTHOR]

Flexible behaviour allows organisms to respond appropriately to changing environmental and social conditions. In the subsocial beetle Nicrophorus vespilloides, females tolerate conspecifics when mating, become aggressive when defending resources, and return to social tolerance when transitioning to parenting. Given the association between octopamine and aggression in insects, we hypothesized that genes in the octopaminergic system would be differentially expressed across different social and reproductive contexts. To test this in N. vespilloides, we first obtained the sequences of orthologues of the synthetic enzymes and receptors of the octopaminergic system. We next compared relative gene expression from virgin females, mated females, mated females alone on a resource required for reproduction and mated females on a resource with a male. Expression varied for five receptor genes. The expression of octopamine β receptor 1 and octopamine β receptor 2 was relatively higher in mated females than in other social conditions. Octopamine β receptor 3 was influenced by the presence or absence of a resource and less by social environment. Octopamine α receptor and octopamine/tyramine receptor 1 gene expression was relatively lower in the mated females with a resource and a male. We suggest that in N. vespilloides the octopaminergic system is associated with the expression of resource defence, alternative mating tactics, social tolerance and indirect parental care. [ABSTRACT FROM AUTHOR]

A queen honey bee mates at ∼6 days of age, storing the sperm in her spermatheca for life. Mating is associated with profound changes in the behaviour and physiology of the queen but the mechanisms underlying these changes are poorly understood. What is known is that the presence of semen in the oviducts and spermatheca is insufficient to initiate laying, and that copulation or CO2 narcosis is necessary for ovary activation. In this study we use real-time quantitative PCR to investigate the expression of biogenic amine receptor genes in the brain and ovarian tissue of queens in relation to their reproductive status. We show that dopamine, octopamine and serotonin receptor genes are expressed in the ovaries of queens, and that natural mating, CO2 narcosis, and the presence of semen in the spermatheca differentially affect their expression. We suggest that these changes may be central to the hormonal cascades that are necessary to initiate oogenesis. [ABSTRACT FROM AUTHOR]

To elucidate the molecular mechanisms underlying honeybee social behaviours, we identified a novel gene, Nb-1, whose expression in the worker brain changes according to the age-dependent division of labour in normal colonies. The open reading frames contained in the Nb-1 cDNA were not conserved in the homologue of a related species, suggesting that the Nb-1 gene product is a non-coding RNA. The distribution of Nb-1-expressing cells partially overlapped that of octopamine-immunoreactive cells and neurosecretory cells, the latter of which are involved in the synthesis and secretion of juvenile hormone (JH). Octopamine and JH control worker task transition, and thus Nb-1 might be involved in task transition through the modulation of octopamine/JH synthesis and secretion. [ABSTRACT FROM AUTHOR]

A cDNA encoding an octopamine (OA) receptor (BmOAR1) was isolated from the nerve tissue of silkworm ( Bombyx mori) larvae. Comparison of amino acid sequences showed that BmOAR1 is highly identical to OA receptors isolated from Periplaneta americana (Pa oa1), Apis mellifera (AmOA1), and Drosophila melanogaster (OAMB or DmOA1A). BmOAR1 was stably expressed in HEK-293 cells. OA above 1 µm led to an increase in intracellular cyclic AMP concentration ([cAMP]i). The synthetic OA-receptor agonist demethylchlordimeform also elevated [cAMP]i to the same maximal level (≈ 5-fold over the basal level) as that induced by OA. However, other biogenic amines, tyramine and dopamine, and chlordimeform were without effects. The [cAMP]i level raised by OA was lowered by antagonists; the rank order of antagonist activity was chlorpromazine > mianserin = yohimbine. Cyproheptadine and metoclopramide had little effect. OA above 100 nm induced a transient or sustained increase in intracellular Ca2+ concentration ([Ca2+]i), depending on the concentration of OA. Sequence homology and functional analysis data indicate that BmOAR1 is an α-adrenergic-like OA receptor of B. mori. [ABSTRACT FROM AUTHOR]

To identify amino acid residues interacting with tyramine (TA) in theBombyx moriTA (BmTA) receptor, several mutant receptors were expressed in HEK-293 cells and examined for their abilities to bind TA and to attenuate forskolin-stimulated cAMP production in response to TA. The D134A BmTA receptor showed no specific[3H]TA binding and no TA-attenuation of cAMP levels. Although the S218A and S222A BmTA receptors showed no specific[3H]TA binding, they still had the ability to mediate the attenuation of cAMP levels in response to the high concentration (100 µm) of TA. The double mutation of Ser218 and Ser222 to Ala, however, led to the loss of TA-attenuation of cAMP levels. The present study thus confirms that at least three amino acid residues play key roles in interaction with TA in the BmTA receptor. [ABSTRACT FROM AUTHOR]

Abstract A cDNA encoding a biogenic amine receptor (B96Bom) was isolated from silkworm (Bombyx mori ) larvae, and the ligand response of the receptor stably expressed in HEK-293 cells was examined. Tyramine (TA) at 0.1–100 µm reduced forskolin (10 µm)-stimulated intracellular cAMP levels by approximately 40%. The inhibitory effect of TA at 1 µm was abolished by yohimbine and chlorpromazine (each 10 µm). Although octopamine (OA) also reduced the cAMP levels, the potency was at least two orders of magnitude lower than that of TA. Furthermore, unlabelled TA (IC50 = 5.2 nm) inhibited specific [3 H]TA binding to the membranes of B96Bom-transfected HEK-293 cells more potently than did OA (IC50 = 1.4 µm) and dopamine (IC50 = 1.7 µm). Taken together with the result of phylogenetic analysis, these findings indicate that the B96Bom receptor is a B. mori TA receptor, which is negatively coupled to adenylate cyclase. The use of this expression system should facilitate physiological studies of TA receptors as well as structure–activity studies of TA receptor ligands. [ABSTRACT FROM AUTHOR]

The biogenic amine receptor genes constitute an ancient and highly divergent family within the larger superfamily of G-protein-coupled receptors. These receptors play a central role in modulating nerve cell activity and thus behaviour. Because the honey bee offers numerous advantages for behavioural studies we endeavoured to isolate as many members of this gene family as possible from the bee. We compared numerous approaches to gene isolation and found that PCR amplification from small subfractions of cDNA or genomic DNA libraries enabled us to isolate clones that are otherwise undetectable. In total we isolated seven biogenic amine receptor clones and identified five additional related sequences by low-stringency Southern hybridization. Two clones, AmBAR4 and AmBAR6, are 84% and 72% identical to the Drosophila 5-HT2 and D1b receptors, respectively, and probably represent orthologous genes. Phylogenetic analysis indicates that AmBAR5 clusters loosely with a variety of tyramine and octopamine receptors with which it shares <66% identity. The other four clones, AmBAR1, AmBAR2, AmBAR3 and AmBAR7, are weakly to moderately related (28–45% identical) to Drosophila dopaminergic or mammalian adrenergic receptors and probably represent receptors of these classes whose orthologues have not previously been isolated from any insect. The honey bee clones expand the size of the known insect biogenic amine receptor gene family to sixteen members. Therefore the size of the biogenic amine receptor gene family of insects approaches that of vertebrates. This is true despite the reduced behavioural and genetic complexity of the insects relative to vertebrate animals. [ABSTRACT FROM AUTHOR]

We have previously reported the cloning of a 5-hydroxytryptamine receptor (Aedes 5-HT7-like receptor) from adult Aedes aegypti. For functional expression of the Aedes 5-HT7-like receptor, CHO-K1 cells were stably transfected with a receptor expression construct, pC5-HT7. The Aedes 5-HT7-like receptor positively coupled to Gs protein, increasing intracellular cAMP in response to 5-HT; adenylyl cyclase activity was induced in a concentration-dependent, saturable manner. Only 5-HT, and not octopamine, dopamine or tyramine, caused the induction of cAMP. At 10 nM 5-HT a weak synergism was observed between octopamine and 5-HT. Other known agonists of the mammalian 5-HT7 receptor were tested. Their order of potency was: 5-HT >> 5-CT = 8-OH-DPAT >> pimozide. This is the first report on the functional expression of a mosquito neurohormone receptor.