Insects have sophisticated olfactory systems that enable them to detect and respond to complex exogenous chemical cues. The encoding mechanisms of these chemical signals have been studied both in their peripheral and central nervous systems (CNS). While many neuromodulators have been shown to play significant roles in olfactory processing within the antennal lobes of the brain, their roles in peripheral olfactory sensory systems, such as the antennae, are less understood. This review focuses on the role of serotonin (5-HT) receptor in the locust antenna, specifically the modulatory function of the serotonin receptor2 on odour inputs. We also review recent studies on the modulation of olfaction in the peripheral nervous systems of other insects and discuss potential directions for future research on the role of neuromodulators in insect peripheral olfactory systems.

Introduction:Serotonin (5-hydroxytryptamine; 5-HT) and GABA (γ-aminobutyric acid) are involved in the regulation of behaviors in the central nervous system. However, it remains unclear whether they modulate olfaction in the peripheral nervous system, and how they modulate olfaction.Methods and results:One 5-HT receptor sequence (Lmig5-HT2) and one GABA receptor sequence (LmigGABAb) were identified in locust antennae by transcriptome analysis and polymerase chain reaction experiments. In situ hybridization localized Lmig5-HT2 to accessory cells, while LmigGABAb was localized to olfactory receptor neurons (ORNs) in locust chemosensilla. Single-unit electrophysiological recordings combined with RNA interference (RNAi) experiments indicated ORNs of locusts with knockdown of Lmig5-HT2 (ds-Lmig5-HT2) and LmigGABAb (ds-LmigGABAb) to some odors had significantly higher responses than wild-type and control locusts in the dose-dependent responses. Moreover, the gaps between the responses of ORNs of RNAi ones and those of wild-type and ds-GFP enlarged with an increase in concentrations of odors.Discussion:Taken together, our findings suggest that 5-HT, GABA, and their receptors exist in the insect peripheral nervous system and that they may function as negative feedback to ORNs and contribute to a fine-tuning mechanism for olfaction in the peripheral nervous system.

The auditory system of female crickets allows them to specifically recognize and approach the species-specific male calling song, defined by sound pulses and silent intervals. Auditory brain neurons form a delay-line and coincidence detector network tuned to the pulse period of the male song. We analyzed the impact of changes in pulse duration on the behavior and the responses of the auditory neurons and the network. We confirm that the ascending neuron AN1 and the local neuron LN2 copy the temporal structure of the song. During ongoing long sound pulses, the delay-line neuron LN5 shows additional rebound responses and the coincidence detector neuron LN3 can generate additional bursts of activity, indicating that these may be driven by intrinsic oscillations of the network. Moreover, the response of the feature detector neuron LN4 is shaped by a combination of inhibitory and excitatory synaptic inputs, and LN4 responds even to long sound pulses with a short depolarization and burst of spikes, like to a sound pulse of natural duration. This response property of LN4 indicates a selective auditory pulse duration filter mechanism of the pattern recognition network, which is tuned to the duration of natural pulses. Comparing the tuning of the phonotactic behavior with the tuning of the local auditory brain neurons to the same test patterns, we find no evidence that a modulation of the phonotactic behavior is reflected at the level of the feature detector neurons. This rather suggests that steering to nonattractive pulse patterns is organized at the thoracic level. NEW & NOTEWORTHY Pulse period selectivity has been reported for the cricket delay-line and coincidence detector network, whereas pulse duration selectivity is evident from behavioral tests. Pulses of increasing duration elicit responses in the pattern recognition neurons, which do not parallel the behavioral responses and indicate additional processing mechanisms. Long sound pulses elicit rhythmic rebound activity and additional bursts, whereas the feature detector neuron reveals a pulse duration filter, expanding our understanding of the pattern recognition process.

In the bispotted field cricket auditory pulse pattern recognition of the species-specific calling song is based on a delay-line and coincidence detection network, established by the activity and synaptic connections of only 5 auditory neurons in the brain. To obtain a more detailed understanding of the network and the dynamic of the neural activity over time we analyzed the response properties of these neurons to test patterns, in which the pulse duration was kept constant while the duration of specific pulse intervals was systematically altered. We confirm that the ascending interneuron AN1 and the local interneuron LN2 copy the structure of the pulse pattern, however with limited resolution at short pulse intervals, further evident in downstream neural responses. In the non-spiking delay-line interneuron LN5 during long pulse intervals full-blown rebound potentials develop over a time course of 35-70 ms. LN5 also reveals an overall increase in its membrane potential tuned to chirps of the calling song pulse pattern. This may contribute to the pattern recognition process by driving the activity of the coincidence-detector LN3 and may indicate a further function of the delay-line neuron LN5. The activity of LN3 and of the feature detector LN4 match the tuning of the phonotactic behavior and demonstrate an increasingly sparse coding of the calling song pulse patterns as evident in the response of the feature detector LN4. The circuitry reveals a fundamental mechanism of auditory pattern recognition and demonstrates a principle of neuronal coding.

Although vomiting is commonly recognized as a protective reaction in response to toxic stimuli, the elaborate sensory processes and necessary molecular components are not fully clear, which is due to a lack of appropriate experimental animal models. Vomiting reflex to volatile chemicals renders locust one candidate for vomiting model. Here, we identified a panel of chemical cues that evoked evident vomiting in locust nymphs and demonstrated the selected combinatorial coding strategy that palps but not antennae olfactory receptor neurons (ORNs) employed. Specifically, knocking down individual palp odorant receptors (ORs) such as OR17, OR21, and OR22 attenuated the vomiting intensity evoked by E-2-hexenal and hexanal, while suppression of OR12 and OR22 augmented vomiting to E-2-hexenal and 2-hexanone, respectively. Furthermore, dual-RNAi treatment against OR17 or OR21 together with OR22 resulted in a much lower response intensity than that of individual OR suppression. Furthermore, OR12 was revealed in palp sensilla basiconica (pb) subtype 3 to tune the neuronal decaying activity to E-2-hexenal. Finally, anterograde labeling indicated that palp ORNs primarily projected into the lobus glomerulatus (LG), and the projection neurons (PNs) in the LG further projected into the accessary calyx (ACA). Together, the establishment of an olfaction-inducible vomiting model in locusts deepens the understanding of olfactory coding logics and provides an opportunity to clarify the neural basis underlying animal vomiting.

Directional hearing is crucial for animals depending on acoustic signals to locate a mate. We focused on crickets to explore the reliability of directional information forwarded to the brain by the ascending auditory interneuron AN1, which is crucial for phonotactic behavior. We presented calling song from −45° to +45° in steps of 3° and compared the phonotactic steering of females walking on a trackball with the directional responses of AN1. Forty percent of females showed good steering behavior and changed their walking direction when the speaker passed the body’s longitudinal axis. The bilateral latency difference between right and left AN1 responses was small and may not be reliable for auditory steering. In respect to spike count, all AN1 recordings presented significant bilateral differences for angles larger than ±18°, yet 35% showed a mean significant difference of 1–3 action potentials per chirp when the frontal stimulus deviated by 3° from their length axis. For small angles, some females had a very similar AN1 activity forwarded to the brain, but the accuracy of their steering behavior was substantially different. Our results indicate a correlation between directional steering and the response strength of AN1, especially for large angles. The reliable steering of animals at small angles would have to be based on small bilateral differences of AN1 activity, if AN1 is the only source providing directional information. We discuss whether such bilateral response difference at small angles can provide a reliable measure to generate auditory steering commands descending from the brain, as pattern recognition is intensity independent.NEW & NOTEWORTHY The ascending auditory interneuron AN1 has been implicated in cricket auditory steering, but at small acoustic stimulation angles, it does not provide reliable directional information. We conclude that either the small bilateral auditory activity differences of the AN1 neurons are enhanced to generate reliable descending steering commands or, more likely, directional auditory steering is mediated via a thoracic pathway, as indicated by the reactive steering hypothesis.

We studied bilateral processing in the auditory ON neurons of crickets using reversible cold-deactivation of the hearing organs by means of Peltier elements. Intracellular recordings of the neurons' activity in response to acoustic stimuli were obtained, while either the ipsilateral or the contralateral hearing organ was cold-deactivated. Afferent activity was abolished at a temperature of approximately 10°C. In ON1, contralateral inhibition had no effect on the latency and amplitude of the phasic onset activity, it enhanced the decline of the onset activity and it decreased the subsequent tonic spiking response to acoustic stimuli. As a consequence, the phasic onset activity became more salient and reciprocal inhibition may support the detection of sound pulses. Contralateral inhibition had a significant impact on the tonic ON1 response, in line with its presumed function to enhance the bilateral auditory contrast. In ON2, experiments confirmed a bilateral excitatory input, with the ipsilateral input dominating the response, and no inhibitory coupling between the ON2 neurons.

Aedes aegypti, the principal vector of yellow fever and dengue fever, is responsible for more than 30,000 deaths annually. Compounds such as carbon dioxide, amino acids, fatty acids and other volatile organic compounds (VOCs) have been widely studied for their role in attracting Ae. aegypti to hosts. Many VOCs from humans are produced by associated skin microbiota. Staphyloccocus epidermidis, although not the most abundant bacteria according to surveys of relative 16S ribosomal RNA abundance, commonly occurs on human skin. Bacteria demonstrate population level decision-making through quorum sensing. Many quorum sensing molecules, such as indole, volatilize and become part of the host odor plum. To date, no one has directly demonstrated the link between quorum sensing (i.e., decision-making) by bacteria associated with a host as a factor regulating arthropod vector attraction. This study examined this specific question with regards to S. epidermidis and Ae. aegypti. Pairwise tests were conducted to examine the response of female Ae. aegypti to combinations of tryptic soy broth (TSB) and S. epidermidis wildtype and agr- strains. The agr gene expresses an accessory gene regulator for quorum sensing; therefore, removing this gene inhibits quorum sensing of the bacteria. Differential attractiveness of mosquitoes to the wildtype and agr- strains was observed. Both wildtype and the agr- strain of S. epidermidis with TSB were marginally more attractive to Ae. aegypti than the TSB alone. Most interestingly, the blood-feeder treated with wildtype S. epidermidis/TSB attracted 74% of Ae. aegypti compared to the agr- strain of S. epidermidis/TSB (P ≤ 0.0001). This study is the first to suggest a role for interkingdom communication between host symbiotic bacteria and mosquitoes. This may have implications for mosquito decision-making with regards to host detection, location and acceptance. We speculate that mosquitoes “eavesdrop” on the chemical discussions occurring between host-associated microbes to determine suitability for blood feeding. We believe these data suggest that manipulating quorum sensing by bacteria could serve as a novel approach for reducing mosquito attraction to hosts, or possibly enhancing the trapping of adults at favored oviposition sites.