Silencing the odorant co-receptor (Orco) in Anoplophora glabripennis disrupts responses to pheromones and host volatiles.

Insects rely primarily on a robust and precise olfactory recognition system to detect chemicals and environmental signals. Olfaction is mediated mainly by various odorant receptors (ORs) expressed on olfactory neurons. The odorant co-receptor (Orco) is a highly conserved and obligatory subunit of ORs, and its combination with conventional ORs to form ligand-gated ion channel heterodimeric complexes plays a crucial role in odor recognition. Anoplophora glabripennis Is a major quarantinable pest that affects broadleaved tree species worldwide. Odorant binding proteins (OBPs) and ORs have been identified in the A. glabripennis genome and the binding properties of some OBPs and their cognate ligands have been clarified. The role of the OR-mediated recognition pathway, however, remains largely uncharacterized. Here, we cloned and sequenced the full-length Orco gene sequence of A. glabripennis and performed structural characterization of the protein. We found that AglaOrco has high sequence homology with Orco from other orders of insects, and that it is highly conserved. Spatio-temporal differential expression analysis revealed that AglaOrco is highly expressed in adult antennae, and that expression at the sexually mature stage is significantly higher than at other developmental stages. There was no significant difference in expression between sexes. Silence AglaOrco using RNAi revealed that expression levels of AglaOrco mRNA fell significantly in both males and females at 72 h post-injection of 5 µg of dsOrco, with no obvious effect on expression of most other olfactory-related genes; however, some were up-or downregulated. For example, silenced Orco-expressing males and females showed a significant reduction in antennal potential responses to the odorants 3-carene, Ocimene, and 4-heptyloxy-1-butanol. Overall, the data suggest that AglaOrco plays an important role in mediating olfactory perception in A. glabripennis, and also identifies potential target genes for environmentally friendly pest control strategies.

Effects of Kv1.3 knockout on pyramidal neuron excitability and synaptic plasticity in piriform cortex of mice.

Kv1.3 belongs to the voltage-gated potassium (Kv) channel family, which is widely expressed in the central nervous system and associated with a variety of neuropsychiatric disorders. Kv1.3 is highly expressed in the olfactory bulb and piriform cortex and involved in the process of odor perception and nutrient metabolism in animals. Previous studies have explored the function of Kv1.3 in olfactory bulb, while the role of Kv1.3 in piriform cortex was less known. In this study, we investigated the neuronal changes of piriform cortex and feeding behavior after smell stimulation, thus revealing a link between the olfactory sensation and body weight in Kv1.3 KO mice. Coronal slices including the anterior piriform cortex were prepared, whole-cell recording and Ca2+ imaging of pyramidal neurons were conducted. We showed that the firing frequency evoked by depolarization pulses and Ca2+ influx evoked by high K+ solution were significantly increased in pyramidal neurons of Kv1.3 knockout (KO) mice compared to WT mice. Western blotting and immunofluorescence analyses revealed that the downstream signaling molecules CaMKII and PKCα were activated in piriform cortex of Kv1.3 KO mice. Pyramidal neurons in Kv1.3 KO mice exhibited significantly reduced paired-pulse ratio and increased presynaptic Cav2.1 expression, proving that the presynaptic vesicle release might be elevated by Ca2+ influx. Using Golgi staining, we found significantly increased dendritic spine density of pyramidal neurons in Kv1.3 KO mice, supporting the stronger postsynaptic responses in these neurons. In olfactory recognition and feeding behavior tests, we showed that Kv1.3 conditional knockout or cannula injection of 5-(4-phenoxybutoxy) psoralen, a Kv1.3 channel blocker, in piriform cortex both elevated the olfactory recognition index and altered the feeding behavior in mice. In summary, Kv1.3 is a key molecule in regulating neuronal activity of the piriform cortex, which may lay a foundation for the treatment of diseases related to piriform cortex and olfactory detection.

The Neurotranscriptome of Monochamus alternatus.

The Japanese pine sawyer Monochamus alternatus serves as the primary vector for pine wilt disease, a devastating pine disease that poses a significant threat to the sustainable development of forestry in the Eurasian region. Currently, trap devices based on informational compounds have played a crucial role in monitoring and controlling the M. alternatus population. However, the specific proteins within M. alternatus involved in recognizing the aforementioned informational compounds remain largely unclear. To elucidate the spatiotemporal distribution of M. alternatus chemosensory-related genes, this study conducted neural transcriptome analyses to investigate gene expression patterns in different body parts during the feeding and mating stages of both male and female beetles. The results revealed that 15 genes in the gustatory receptor (GR) gene family exhibited high expression in the mouthparts, most genes in the odorant binding protein (OBP) gene family exhibited high expression across all body parts, 22 genes in the odorant receptor (OR) gene family exhibited high expression in the antennae, a significant number of genes in the chemosensory protein (CSP) and sensory neuron membrane protein (SNMP) gene families exhibited high expression in both the mouthparts and antennae, and 30 genes in the ionotropic receptors (IR) gene family were expressed in the antennae. Through co-expression analyses, it was observed that 34 genes in the IR gene family were co-expressed across the four developmental stages. The Antenna IR subfamily and IR8a/Ir25a subfamily exhibited relatively high expression levels in the antennae, while the Kainate subfamily, NMDA subfamily, and Divergent subfamily exhibited predominantly high expression in the facial region. MalIR33 is expressed only during the feeding stage of M. alternatus, the MalIR37 gene exhibits specific expression in male beetles, the MalIR34 gene exhibits specific expression during the feeding stage in male beetles, the MalIR8 and MalIR39 genes exhibit specific expression during the feeding stage in female beetles, and MalIR8 is expressed only during two developmental stages in male beetles and during the mating stage in female beetles. The IR gene family exhibits gene-specific expression in different spatiotemporal contexts, laying the foundation for the subsequent selection of functional genes and facilitating the full utilization of host plant volatiles and insect sex pheromones, thereby enabling the development of more efficient attractants.

Neuroregulation of foraging behavior mediated by the olfactory co-receptor Orco in termites.

Olfaction is critical for survival because it allows animals to look for food and detect pheromonal cues. Neuropeptides modulate olfaction and behaviors in insects. While how the neuroregulation of olfactory recognition affects foraging behavior in termites is still unclear. Here, we analyzed the change after silencing the olfactory co-receptor gene (Orco) and the neuropeptide Y gene (NPY), and then investigated the impact of olfactory recognition on foraging behavior in Odontotermes formosanus under different predation pressures. The knockdown of Orco resulted in the reduced Orco protein expression in antennae and the decreased EAG response to trail pheromones. In addition, NPY silencing led to the damaged ability of olfactory response through downregulating Orco expression. Both dsOrco- and dsNPY-injected worker termites showed significantly reduced walking activity and foraging success. Additionally, we found that 0.1 pg/cm trail pheromone and nestmate soldiers could provide social buffering to relieve the adverse effect of predator ants on foraging behavior in worker termites with the normal ability of olfactory recognition. Our orthogonal experiments further verified that Orco/NPY genes are essential in manipulating termite olfactory recognition during foraging under different predation pressures, suggesting that the neuroregulation of olfactory recognition plays a crucial role in regulating termite foraging behavior.

Genome-wide identification of candidate chemosensory receptors in the bean bug Riptortus pedestris (Hemiptera: Alydidae) and the functional verification of its odorant receptor co-receptor (Orco) in recognizing aggregation pheromone.

A sophisticated and sensitive olfactory system plays a vital role in the survival and reproduction of insects. Chemosensory receptors are indispensable for the molecular recognition and discrimination of semiochemicals. Riptortus pedestris is a notorious pest of legume plants, resulting in yield losses and quality decreases in soybeans. It is well accepted that R. pedestris highly relies on its olfactory system in detecting aggregation pheromones, host volatiles, and pesticides; however, little research focused on its chemosensory receptors. In the present study, we identified 237 odorant receptors (ORs), 42 gustatory receptors (GRs), and 31 ionotropic receptors (IRs) from the reported genome of R. pedestris, and analyzed their phylogenetic relationship with other hemipteran species. Through the results of RNA-seq and real-time quantitative PCR (qRT-PCR), we found that RpedORs displayed different expression levels in the antennae of R. pedestris at different development stages. To further verify the function of odorant receptor co-receptor (Orco), an obligate and unique insect OR, we silenced RpedOrco by RNA interference (RNAi) method. The results showed that silencing RpedOrco could significantly impair the response to aggregation pheromone in R. pedestris, indicating that RpedOrco plays an essential role in odorant detection. Our results can provide the theoretical foundations for revealing the olfactory recognition mechanism of R. pedestris and help explore and develop novel olfactory-based agents against this pest.

Genome-Wide Identification of the Odorant Receptor Gene Family and Revealing Key Genes Involved in Sexual Communication in Anoplophora glabripennis.

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.

Are you my mummy? Long-term olfactory memory of mother's body odour by offspring in the domestic cat.

Longevity of odour memories, particularly those acquired during early development, has been documented in a wide range of taxa. Here, we report that kittens of the domestic cat retained a memory into adult life of their mother´s body odour experienced before weaning. Kittens from 15 litters were tested when permanently separated from their mother at weaning on postnatal week 8, and tested again when 4 and 6 months and over 1 year of age. When presented with a simultaneous three-way choice between body odour of their own mother, of an unknown female of similar reproductive condition and a blank stimulus, weaning-age kittens sniffed the cotton swab with the odour of an unknown female longer. This preference, however, changed when as adults the subjects sniffed the cotton swab with their own mother's odour longer. We conclude that kittens form a long-lasting memory of the body odour of their mother, and by implication, that mothers retain an individual odour signature sufficiently stable across age and changes in their reproductive state to be distinguishable by their adult offspring. What this means in functional or cognitive terms is not yet clear. Does such "recognition" have a specific biological function and a specific cognitive representation? Or is it rather part of a more general phenomenon well known in (human) olfaction of odours that are familiar generally being judged more pleasant, and that might then influence olfactory-guided behaviour in a variety of contexts?

Characterizing the Role of Orco Gene in Detecting Aggregation Pheromone and Food Resources in Protaetia brevitarsis Leiws (Coleoptera: Scarabaeidae).

An accurate olfactory system for recognizing semiochemicals and environmental chemical signals plays crucial roles in survival and reproduction of insects. Among all olfaction-related proteins, olfactory receptors (ORs) contribute to the conversion of chemical stimuli to electric signals and thereby are vital in odorant recognition. Olfactory receptor co-receptor (Orco), one of the most conserved ORs, is extremely essential in recognizing odorants through forming a ligand-gated ion channel complex with conventional ligand-binding odorant receptors. We have previously identified aggregation pheromone in Protaetia brevitarsis (Coleoptera: Scarabaeidae), a native agricultural and horticultural pest in East-Asia. However, to our best knowledge, its olfaction recognition mechanisms are still veiled. To illustrate how P. brevitarsis recognize aggregation pheromone and host plants, in the present study we cloned and sequenced the full-length Orco gene from P. brevitarsis antennae (named PbreOrco) and found that PbreOrco is highly conserved and similar to Orcos from other Coleoptera insects. Our real-time quantitative PCR (qRT-PCR) results showed that PbreOrco is mainly expressed in antenna. We also demonstrated that silencing PbreOrco using RNA interference through injecting dsOrco fragment significantly inhibited PbreOrco expression in comparison with injecting control dsGFP and subsequently revealed using electroantennogram and behavioral bioassays that decreasing PbreOrco transcript abundance significantly impaired the responses of P. brevitarsis to intraspecific aggregation pheromone and prolonged the time of P. brevitarsis spending on food seeking. Overall, our results demonstrated that PbreOrco is crucial in mediating odorant perception in P. brevitarsis.

Electroantennogram reveals a strong correlation between the passion of honeybee and the properties of the volatile.

INTRODUCTION: Insects use their antennae to detect food, mates, and predators, mainly via olfactory recognition of specific volatile compounds. Honeybees also communicate, learn complex tasks, and show adaptable behavior by recognizing and responding to specific odors. However, the relationship between the electroantennogram and the passion of honeybee has not been determined. METHODS: We established a four-channel maze system to detect the degree of sensitivity of the honeybee's antenna to different odors. In addition, electroantennography (EAG) signal was recorded from the right antennae of the honeybees in our experiments to explore electrophysiological responses to different volatiles. RESULTS: The olfactory sensilla on the antennae of honeybees engender distinct electrophysiological responses to different volatiles. The bees were exposed to honey, 1-hexanol and formic acid, and EAG parameters like depolarization time, falling slope, and amplitude were measured. The EAG indicators varied significantly between honey and formic acid, indicating either "happy" or "anxious" moods. CONCLUSIONS: Honeybee can express its passion by the characteristic changes of EAG parameters. We defined a preference factor (F) to quantify the preference of bees to varying concentrations of different compounds, where greater positive values indicate an increased passion. Our findings provide novel insights into the understanding of odor recognition in insects.

Chemical Profiles of Integumentary and Glandular Substrates in Australian Sea Lion Pups (Neophoca cinerea).

Recognition of individuals or classes of individuals plays an important role in the communication systems of many mammals. The ability of otariid (i.e., fur seal and sea lion) females to locate and identify their offspring in colonies after returning from regular foraging trips is essential to successful pup rearing. It has been shown that olfaction is used to confirm the identity of the pup by the mother when they reunite, yet the processes by which this chemical recognition occurs remain unclear. Using gas chromatography-mass spectrometry, we examined chemical profiles of integumentary and glandular secretions/excretions from pre- and post-molt Australian sea lion pups (Neophoca cinerea) and compared fur and swab samples to assess data collection methods. Multivariate statistics were applied to assess differences in chemical composition between body regions and sexes. We found differences among secretions from various body regions, driven by the distinctiveness of the oral odor mixture. The fine-scale trends in pre- and post-molt pups seem to differ due to changes in the behavior of pups and consequent decrease in the transfer of compounds among adjacent body regions in older pups. Volatile compounds from exocrine substrates were not distinct for different sexes. We also show that swab samples provide better data for exploring social olfaction than fur samples for this species. Obtaining fundamental chemical information, in this case chemical profiles of animals, and discerning differences in chemical composition is an important step toward fully exploring the intricacies of mother-offspring olfactory recognition and its underlying processes.

Olfactory Memory Impairment Differs by Sex in a Rodent Model of Pediatric Radiotherapy.

Although an effective treatment for pediatric brain tumors, cranial radiation therapy (CRT) damages surrounding healthy tissue, thereby disrupting brain development. Animal models of pediatric CRT have primarily relied on visual tasks to assess cognitive impairment. Moreover, there has been a lack of sex comparisons as most research on the cognitive effects of pediatric CRT does not include females. Therefore, we utilized olfaction, an ethologically relevant sensory modality, to assess cognitive impairment in an animal model of CRT that included both male and female mice. Specifically, we used the novel odor recognition (NOdorR) task with social odors to test recognition memory, a cognitive parameter that has been associated with olfactory neurogenesis, a form of cellular plasticity damaged by CRT. In addition to odor recognition memory, olfactory ability or discrimination of non-social and social odors were assessed both acutely and 3 months after CRT. Magnetic resonance imaging (MRI) and histology were performed after behavioral testing to assess long-term damage by CRT. Long-term but not acute radiation-induced impairment in odor recognition memory was observed, consistent with delayed onset of cognitive impairment in human patients. Males showed greater exploration of social odors than females, but general exploration was not affected by irradiation. However, irradiated males had impaired odor recognition memory in adulthood, compared to non-irradiated males (or simply male controls). Female olfactory recognition memory, in contrast, was dependent on estrus stage. CRT damage was demonstrated by (1) histological evaluation of olfactory neurogenesis, which suggested a reduction in CRT versus control, and (2) imaging analyses which showed that the majority of brain regions were reduced in volume by CRT. Specifically, two regions involved in social odor processing (amygdala and piriform cortex) were damaged by cranial irradiation in males but not females, paralleling olfactory recognition findings.

A cytochrome P450 gene plays a role in the recognition of sex pheromones in the tobacco cutworm, Spodoptera litura.

Cytochrome P450 (P450 or CYP) genes are involved in fundamental physiological functions, and might be also associated with the olfactory recognition of sex pheromones in beetles and moths. A P450 gene, Spodoptera litura CYP4L4 (SlituCYP4L4), was cloned for the first time from the antennae of S. litura. SlituCYP4L4 was almost exclusively expressed in the adult stage and predominantly expressed in the adult antennae. In situ hybridization showed that SlituCYP4L4 localized mainly at the base of the long sensilla trichoidea, which responds to sex pheromone components. Pretreatment with an S. litura sex pheromone significantly reduced the expression levels of SlituCYP4L4, consistent with other genes involved in sex pheromone recognition. The expression level of SlituCYP4L4 was different in moths collected with different ratios of sex pheromone lures and collected in different geographical locations. After gene knockdown of SlituCYP4L4 in the antennae, the electroantennogram (EAG) responses of male and female moths to (9Z,11E)-tetradecadienyl acetate or (9Z,12E)-tetradecadienyl acetate were significantly decreased. In contrast, EAG responses to plant volatiles and sex pheromones of other moth species were not significantly influenced in these moths. SlituCYP4L4 was also expressed in the gustatory tissues and sensilla, which suggests that SlituCYP4L4 may have other functions in the chemosensory system. Our results have shown for the first time the function of a CYP gene with appendage-specific expression in insect sex pheromone recognition, especially in adult moths.

Olfactory recognition memory is disrupted in young mice with chronic low-level lead exposure.

Chronic developmental lead exposure yielding very low blood lead burden is an unresolved child public health problem. Few studies have attempted to model neurobehavioral changes in young animals following very low level exposure, and studies are needed to identify tests that are sensitive to the neurobehavioral changes that may occur. Mechanisms of action are not yet known however results have suggested that hippocampus/dentate gyrus may be uniquely vulnerable to early chronic low-level lead exposure. This study examined the sensitivity of a novel odor recognition task to differences in pre-adolescent C57BL/6J mice chronically exposed from birth to PND 28, to 0 ppm (control), 30 ppm (low-dose), or 330 ppm (higher-dose) lead acetate (N=33). Blood lead levels (BLLs) determined by ICP-MS ranged from 0.02 to 20.31 µg/dL. Generalized linear mixed model analyses with litter as a random effect showed a significant interaction of BLL×sex. As BLLs increased olfactory recognition memory decreased in males. Among females, non-linear effects were observed at lower but not higher levels of lead exposure. The novel odor detection task is sensitive to effects associated with early chronic low-level lead exposure in young C57BL/6J mice.

Markers of brain illness may be hidden in your olfactory ability: a Japanese perspective.

There is evidence that impaired human cognitive abilities are reflected by loss of olfactory abilities. Declining olfactory perception may be a biomarker for impairment of cognitive function and of impending neurogenerative disorders. As olfactory perception may differ between culture and ethnic group, we sought to confirm this relationship with Japanese participants. In this study, we examined possible relationships between age and olfactory abilities in healthy Japanese subjects (control subjects) over a wide range of ages and compared this relationship with that observed in three neurodegenerative disorders; patients with Parkinson's disease (PD), Type 1 myotonic dystrophy (DM1) and Alzheimer's disease (AD). In control subjects, both threshold and recognition abilities decreased with age. Ability to detect odors was generally intact in most control subjects, however, we found that the abilities of individuals in the three different patient populations to recognize odors were impaired relative to control subjects. All three types of patients exhibited decreased or impaired odor-recognition compared with age-matched controls. Previous studies showed the causes of olfactory impairments in PD and AD patients were attributable to pathological changes and MRI signal abnormalities in limbic areas, including the amygdala (AMG), entorhinal cortex (ENT), hippocampus (HI), and orbitofrontal cortex (OFC). Another study reported that DM1 patients have bilateral lesions in anterior temporal areas, including the subcortical white matter, AMG, ENT and insula. Our findings underscore the need to pay careful attention to significant decreases of odor identification abilities caused by diverse forms of abnormal brain function, especially in the AMG, ENT and HI.