Role In Olfactory Signal Transduction Research Articles

Candidate odorant-binding protein and chemosensory protein genes in the turnip aphid Lipaphis erysimi.

The turnip aphid, Lipaphis erysimi Kaltenbach, inflicts heavy damage on cruciferous crops worldwide. In these insects, olfactory perception is crucial for mating, host location, and oviposition. Both odorant-binding proteins (OBPs) and chemosensory proteins (CSPs) are responsible for the delivery of host odorants and pheromones during initial molecular interactions. In this study, antennal and body transcriptomes of L. erysimi were generated through the deep sequencing of RNA libraries. A dataset of 11 LeryOBP and four LeryCSP transcripts was identified among assembled unigenes and subjected to sequence analysis. Phylogenetic analysis found a one-to-one orthologous relationship between LeryOBP/LeryCSP and its corresponding homologs from other aphid species. Further quantitative real-time PCR analyses across developmental stages and tissues showed that five LeryOBP genes (i.e., LeryGOBP, LeryOBP6, LeryOBP7, LeryOBP9, and LeryOBP13) and LeryCSP10 were specifically or significantly elevated in the antennae compared with other tissues. Moreover, two transcripts (i.e., LeryGOBP and LeryOBP6) exhibited remarkably higher expression levels in alate aphids, implying their potentially functional role in the perception of new host plant locations. These results present the identification and expression of OBP/CSP genes in L. erysimi, providing valuable insights into their putative role in olfactory signal transduction.

Type 3 adenylyl cyclase in the main olfactory epithelium participates in depression-like and anxiety-like behaviours

BackgroundDeficiency of olfaction is thought to be associated with depression, and type 3 adenylyl cyclase (AC3) genetic knockout and forebrain knockout mice show depression-like behaviours. AC3 is expressed in the main olfactory epithelium (MOE) and hippocampus, which plays an important role in olfactory signal transduction. However, it is unclear whether AC3 in the MOE also plays a role in the pathogenesis of depression. Thus, we aimed to study the relationship between AC3 in the MOE and the pathogenesis of depression. MethodsWe obtained anosmic mice by intranasal perfusion of zinc sulphate (ZnSO4) (ZnSO4 mice), and distinctively knocked down AC3 in the MOE (AC3KD/MOE mice) by CRISPR/cas9 technology. Behavioural tests related to depression and anxiety were employed to evaluate the depression- and anxiety-like behaviours of mice. The mRNA and protein expressions of tyrosine hydroxylase (TH), dopamine receptors (Drds), and N-Methyl D-aspartate receptor subunit 2B (GluN2B) in the hippocampus of mice were investigated by qPCR and western blotting to explore the mechanism of depression and anxiety caused by AC3 in the MOE, preliminarily. ResultsCompared with NaCl mice, ZnSO4 mice exhibited depression-like behaviours in tail suspension tests (TST), forced swimming tests, and social (FST) interaction tests (SIT), but showed no anxiety-like behaviours in anxiety-related behavioural tests. The mRNA and protein expressions of Drd3 and GluN2B in the hippocampus of ZnSO4 mice were significantly downregulated. Compared with the negative control mice (NC mice), AC3KD / MOE mice showed depression-like behaviours in TST, FST, and SIT tests, anxiety-like behaviours in light/dark transition test, elevated-plus maze test, and novelty-suppressed feeding test. The protein expressions of Drd3, TH, and GluN2B were significantly downregulated in the hippocampus. LimitationsWe did not further demonstrate that AC3 in the MOE causes depression through the dopaminergic nervous system with dopamine or dopamine receptor agonists. ConclusionsOur data demonstrate that intranasal infusion of ZnSO4 can cause depression-like behaviours and has no effect on anxiety-like behaviours. Specific knockdown of AC3 in the MOE can cause depression-like and anxiety-like behaviours. The behavioural changes caused by intranasal ZnSO4 and specific knockdown of AC3 in the MOE can be related to the significant downregulation of dopaminergic system and GluN2B expressions in the hippocampus of mice.

Intranasal sodium citrate solution improves olfaction in post-viral hyposmia

Calcium plays an integral role in olfactory signal transduction, including feedback inhibition. Sodium citrate acts as a calcium sequestrant and when applied intranasally, reduces free calcium available for feedback inhibition, which should theoretically improve olfaction. We aimed to investigate the utility of intranasal sodium citrate in improving the olfactory function of hyposmic patients, by performing this prospective placebo controlled, single-blind trial. Monorhinal olfactory testing for odour identification and threshold was performed in hyposmic patients using Sniffin Sticks, before and after treatment. Treatment consisted of one-off sodium citrate solution application to the olfactory cleft. Sodium chloride solution was applied to the contralateral olfactory cleft, which therefore acted as placebo control. Patients were blinded to the side of sodium citrate application, and side of treatment was randomized between patients. 57 patients participated, aged 22-79. Causes of hyposmia included: post-viral (7); posttraumatic (10); sinonasal disease (30) and idiopathic (10). Compared with placebo, there was significant improvement in the identification scores of participants with post-viral hyposmia, following sodium citrate treatment. No significant change in olfactory function occurred for either identification or threshold in any other aetiological subgroup. Intranasal sodium citrate may be of benefit to patients with post-viral hyposmia.

Intranasal sodium citrate solution improves olfaction in post-viral hyposmia.

Calcium plays an integral role in olfactory signal transduction, including feedback inhibition. Sodium citrate acts as a calcium sequestrant and when applied intranasally, reduces free calcium available for feedback inhibition, which should theoretically improve olfaction. We aimed to investigate the utility of intranasal sodium citrate in improving the olfactory function of hyposmic patients, by performing this prospective placebo controlled, single-blind trial. Monorhinal olfactory testing for odour identification and threshold was performed in hyposmic patients using Sniffin Sticks, before and after treatment. Treatment consisted of one-off sodium citrate solution application to the olfactory cleft. Sodium chloride solution was applied to the contralateral olfactory cleft, which therefore acted as placebo control. Patients were blinded to the side of sodium citrate application, and side of treatment was randomized between patients. 57 patients participated, aged 22-79. Causes of hyposmia included: post-viral (7); posttraumatic (10); sinonasal disease (30) and idiopathic (10). Compared with placebo, there was significant improvement in the identification scores of participants with post-viral hyposmia, following sodium citrate treatment. No significant change in olfactory function occurred for either identification or threshold in any other aetiological subgroup. Intranasal sodium citrate may be of benefit to patients with post-viral hyposmia.

Adenylyl cyclase 3 haploinsufficiency confers susceptibility to diet-induced obesity and insulin resistance in mice.

Adenylyl cyclase 3 (Adcy3), a member of the mammalian adenylyl cyclase family responsible for generating the second messenger cAMP, has long been known to play an essential role in olfactory signal transduction. Here, we demonstrated that Adcy3 heterozygous null mice displayed increased visceral adiposity in the absence of hyperphagia and developed abnormal metabolic features characterized by impaired insulin sensitivity, dyslipidemia, and increased plasma levels of proinflammatory cytokines on both chow and high-fat diet (HFD). Of note, HFD decreased the Adcy3 expression in white adipose tissue, liver, and muscle. We also report for the first time that Adcy3 haploinsufficiency resulted in reduced expression of genes involved in thermogenesis, fatty acid oxidation, and insulin signaling, with enhanced expression of genes related to adipogenesis in peripheral tissues of mice. In conclusion, these findings suggest that cAMP signals generated by Adcy3 in peripheral tissues may play a pivotal role in modulating obesity and insulin sensitivity.

The first mutation in CNGA2 in two brothers with anosmia.

Isolated congenital anosmia (ICA) is a rare disorder, where otherwise healthy individuals present with an inability to smell since birth. A list of studies have described the genes involved in syndromic anosmia; however, the genetics of ICA is still in its infancy. Studies in mice show that the cyclic nucleotide-gated channel subunit CNGA2, expressed in the olfactory epithelium has a crucial role in olfactory signal transduction. We have identified a novel X-linked stop mutation in CNGA2 (c.634C>T, p.R212*) in two brothers with ICA using exome sequencing. No additional mutations in CNGA2 were identified in a cohort of 31 non-related ICA individuals. Magnetic resonance brain imaging revealed diminished olfactory bulbs and flattened olfactory sulci. This is the first report of a mutation in the cyclic nucleotide-gated gene CNGA2 and supports the critical role of this gene in human olfaction.

Developmental expression of the calcium‐activated chloride channels TMEM16A and TMEM16B in the mouse olfactory epithelium

Calcium-activated chloride channels are involved in several physiological processes including olfactory perception. TMEM16A and TMEM16B, members of the transmembrane protein 16 family (TMEM16), are responsible for calcium-activated chloride currents in several cells. Both are present in the olfactory epithelium of adult mice, but little is known about their expression during embryonic development. Using immunohistochemistry we studied their expression in the mouse olfactory epithelium at various stages of prenatal development from embryonic day (E) 12.5 to E18.5 as well as in postnatal mice. At E12.5, TMEM16A immunoreactivity was present at the apical surface of the entire olfactory epithelium, but from E16.5 became restricted to a region near the transition zone with the respiratory epithelium, where localized at the apical part of supporting cells and in their microvilli. In contrast, TMEM16B immunoreactivity was present at E14.5 at the apical surface of the entire olfactory epithelium, increased in subsequent days, and localized to the cilia of mature olfactory sensory neurons. These data suggest different functional roles for TMEM16A and TMEM16B in the developing as well as in the postnatal olfactory epithelium. The presence of TMEM16A at the apical part and in microvilli of supporting cells is consistent with a role in the regulation of the chloride ionic composition of the mucus covering the apical surface of the olfactory epithelium, whereas the localization of TMEM16B to the cilia of mature olfactory sensory neurons is consistent with a role in olfactory signal transduction.

A functional role for Anopheles gambiae Arrestin1 in olfactory signal transduction

Insect sensory arrestins act to desensitize visual and olfactory signal transduction pathways, as evidenced by the phenotypic effects of mutations in the genes encoding both Arr1 and Arr2 in Drosophila melanogaster. To assess whether such arrestins play similar roles in other, more medically relevant dipterans, we examined the ability of Anopheles gambiae sensory arrestin homologs AgArr1 and AgArr2 to rescue phenotypes associated with an olfactory deficit observed in D. melanogaster arrestin mutants. Of these, only AgArr1 facilitated significant phenotypic rescue of the corresponding Drosophila arr mutant olfactory phenotype, consistent with the view that functional orthology is shared by these Arr1 homologs. These results represent the first step in the functional characterization of AgArr1, which is highly expressed in olfactory appendages of An. gambiae in which it is likely to play an essential role in olfactory signal transduction. In addition to providing insight into the common elements of the peripheral olfactory system of dipterans, this work validates the importance of AgArr1 as a potential target for novel anti-malaria strategies that focus on olfactory-based behaviors in An. gambiae.

Calmodulin-regulated adenylyl cyclases: cross-talk and plasticity in the central nervous system.

Gene disruption studies have shown that the Ca(2+)-stimulated adenylyl cyclases, AC1 and AC8, are critical for some forms of synaptic plasticity, including long-term potentiation as well as long-term memory formation (LTM). It is hypothesized that these enzymes are required for LTM to support the increased expression of a family of genes regulated through the cAMP/Ca(2+) response element-binding protein/cAMP response element transcriptional pathway. In contrast to AC1 and AC8, AC3 is a Ca(2+)-inhibited adenylyl cyclase that plays an essential role in olfactory signal transduction. Coupling of odorant receptors to AC3 stimulates cAMP transients that function as the major second messenger for olfactory signaling. These cAMP transients are caused, at least in part, by Ca(2+) inhibition of AC3, which is mediated through calmodulin-dependent protein kinase II. The unique structure and regulatory properties of these adenylyl cyclases make them attractive drug target sites for modulation of a number of physiological processes including memory formation and olfaction.

Amplification of odor-induced Ca(2+) transients by store-operated Ca(2+) release and its role in olfactory signal transduction.

A critical role of Ca(2+) in vertebrate olfactory receptor neurons (ORNs) is to couple odor-induced excitation to intracellular feedback pathways that are responsible for the regulation of the sensitivity of the sense of smell, but the role of intracellular Ca(2+) stores in this process remains unclear. Using confocal Ca(2+) imaging and perforated patch recording, we show that salamander ORNs contain a releasable pool of Ca(2+) that can be discharged at rest by the SERCA inhibitor thapsigargin and the ryanodine receptor agonist caffeine. The Ca(2+) stores are spatially restricted; emptying produces compartmentalized Ca(2+) release and capacitative-like Ca(2+) entry in the dendrite and soma but not in the cilia, the site of odor transduction. We deplete the stores to show that odor stimulation causes store-dependent Ca(2+) mobilization. This odor-induced Ca(2+) release does not seem to be necessary for generation of an immediate electrophysiological response, nor does it contribute significantly to the Ca(2+) transients in the olfactory cilia. Rather, it is important for amplifying the magnitude and duration of Ca(2+) transients in the dendrite and soma and is thus necessary for the spread of an odor-induced Ca(2+) wave from the cilia to the soma. We show that this amplification process depends on Ca(2+)-induced Ca(2+) release. The results indicate that stimulation of ORNs with odorants can produce Ca(2+) mobilization from intracellular stores without an immediate effect on the receptor potential. Odor-induced, store-dependent Ca(2+) mobilization may be part of a feedback pathway by which information is transferred from the distal dendrite of an ORN to its soma.

Localization of the olfactory cyclic nucleotide-gated channel subunit 1 in normal, embryonic and regenerating olfactory epithelium

The spatial and temporal expression of subunit 1 of the olfactory cyclic nucleotide-gated channel was investigated using affinity-purified anti-fusion protein antibodies. Immunoreactivity was most prominent in the ciliary layer of the olfactory epithelium, but high protein expression was also seen along the entire length of olfactory receptor neuronal axons to the level of the glomeruli. Electron microscopy showed that the long, thin distal compartments of olfactory cilia labeled more prominently than their thicker proximal segments. This was true as soon as these distal parts began to develop. Using light microscopy, developmental expression of olfactory cyclic nucleotide-gated channel subunit 1 could be detected in discrete populations of olfactory receptor neurons by embryonic day 14. Other signaling molecules are expressed either later (G olf) or only at the level of the epithelial surface and not in axons (adenylyl cyclase type III). Following unilateral lesions of the olfactory bulb, olfactory cyclic nucleotide-gated channel subunit 1 immunoreactivity was present early and throughout developing olfactory receptor neurons; adenylyl cyclase type III immunoreactivity, in contrast, was detectable only later, and again present only in the cilial layer. These results support the hypothesis that this subunit of the olfactory cyclic nucleotide-gated channel may be involved in olfactory axon guidance, in addition to its well-described role in olfactory signal transduction.

Topochemistry of moth olfactory sensilla

Fine structure immunocytochemistry permitted the localization of proteins, which are presumably relevant to olfactory signal transduction in the olfactory sensilla of the silkmoths Bombyx mori (Lepidoptera: Bombycidae), Antheraea pernyi and Antheraea polyphemus (Lepidoptera: Saturniidae). Odorant-binding proteins of different classes, pheromone-binding protein and general odorant-binding protein 2, were localized in different olfactory sensilla. The expression pattern of these proteins among the population of olfactory sensilla was analysed and correlated with the olfactory specificity of sensilla. The results support the notion that odorant-binding proteins may play a multifunctional role in olfactory signal transduction. Further immunocytochemical experiments concerned the intracellular signalling pathways. A G-protein of the G q-family and an inositol 1,4,5-trisphosphate-receptor, which is an inositol 1,4,5-trisphosphate-dependent Ca 2+ —channel, were localized in the receptive dendrites of olfactory receptor cells and other parts of the neuroepithelium. Both molecules are involved in the inositol 1,4, 5-trisphosphate/diacylglycerol second messenger pathway, which is supposed to mediate olfactory signal transduction in insects. Calmodulin, an ubiquitous calcium-binding protein, and calcineurin, a Ca 2+/calmodulin-regulated phosphatase, were also localized at high labelling densities in the dendrites of olfactory receptor cells. This co-expression and the finding that a Ca 2+-channel (the inositol 1,4,5-trisphosphate-receptor) is localized in the receptor cell dendrites supports the notion that calcium plays a role in olfactory signal transduction. Finally, a possible nitric oxide-synthase was localized in the haemolymph, which suggests that the localized form of the enzyme has no functional relevance for olfactory signal transduction.