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      Candidate odorant binding proteins and chemosensory proteins in the larval chemosensory tissues of two closely related noctuidae moths, Helicoverpa armigera and H. assulta

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          Abstract

          In order to acquire enough nutrients and energy for further development, larvae need to invest a large portion of their sensory equipments to identify food sources. Yet, the molecular basis of odor-driven behavior in larvae has been poorly investigated. Information on olfactory genes, particularly odorant binding proteins (OBPs) and chemosensory proteins (CSPs) which are involved in the initial steps of olfaction is very scarce. In this study, we have identified 26 OBP and 21 CSP genes from the transcriptomes of Helicoverpa armigera larval antennae and mouthparts. A comparison with the 34 OBP and 18 CSP genes of the adult antenna, revealed four novel OBPs and seven novel CSPs. Similarly, 27 OBPs (six novel OBPs) and 20 CSPs (6 novel CSPs) were identified in the transcriptomes of Helicoverpa assulta larval antennae and mouthparts. Tissue-specific profiles of these soluble proteins in H. armigera showed that 6 OBP and 4 CSP genes are larval tissue-specific, 15 OBPs and 13 CSPs are expressed in both larvae and adult, while the rest are adult- specific. Our data provide useful information for functional studies of genes involved in larval foraging.

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          Insect olfactory receptors are heteromeric ligand-gated ion channels.

          In insects, each olfactory sensory neuron expresses between one and three ligand-binding members of the olfactory receptor (OR) gene family, along with the highly conserved and broadly expressed Or83b co-receptor. The functional insect OR consists of a heteromeric complex of unknown stoichiometry but comprising at least one variable odorant-binding subunit and one constant Or83b family subunit. Insect ORs lack homology to G-protein-coupled chemosensory receptors in vertebrates and possess a distinct seven-transmembrane topology with the amino terminus located intracellularly. Here we provide evidence that heteromeric insect ORs comprise a new class of ligand-activated non-selective cation channels. Heterologous cells expressing silkmoth, fruitfly or mosquito heteromeric OR complexes showed extracellular Ca2+ influx and cation-non-selective ion conductance on stimulation with odorant. Odour-evoked OR currents are independent of known G-protein-coupled second messenger pathways. The fast response kinetics and OR-subunit-dependent K+ ion selectivity of the insect OR complex support the hypothesis that the complex between OR and Or83b itself confers channel activity. Direct evidence for odorant-gated channels was obtained by outside-out patch-clamp recording of Xenopus oocyte and HEK293T cell membranes expressing insect OR complexes. The ligand-gated ion channel formed by an insect OR complex seems to be the basis for a unique strategy that insects have acquired to respond to the olfactory environment.
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            Drosophila odorant receptors are both ligand-gated and cyclic-nucleotide-activated cation channels.

            From worm to man, many odorant signals are perceived by the binding of volatile ligands to odorant receptors that belong to the G-protein-coupled receptor (GPCR) family. They couple to heterotrimeric G-proteins, most of which induce cAMP production. This second messenger then activates cyclic-nucleotide-gated ion channels to depolarize the olfactory receptor neuron, thus providing a signal for further neuronal processing. Recent findings, however, have challenged this concept of odorant signal transduction in insects, because their odorant receptors, which lack any sequence similarity to other GPCRs, are composed of conventional odorant receptors (for example, Or22a), dimerized with a ubiquitously expressed chaperone protein, such as Or83b in Drosophila. Or83b has a structure akin to GPCRs, but has an inverted orientation in the plasma membrane. However, G proteins are expressed in insect olfactory receptor neurons, and olfactory perception is modified by mutations affecting the cAMP transduction pathway. Here we show that application of odorants to mammalian cells co-expressing Or22a and Or83b results in non-selective cation currents activated by means of an ionotropic and a metabotropic pathway, and a subsequent increase in the intracellular Ca(2+) concentration. Expression of Or83b alone leads to functional ion channels not directly responding to odorants, but being directly activated by intracellular cAMP or cGMP. Insect odorant receptors thus form ligand-gated channels as well as complexes of odorant-sensing units and cyclic-nucleotide-activated non-selective cation channels. Thereby, they provide rapid and transient as well as sensitive and prolonged odorant signalling.
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              Evolutionary ecology of the relationship between oviposition preference and performance of offspring in phytophagous insects

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                Author and article information

                Contributors
                Role: Editor
                Journal
                PLoS One
                PLoS ONE
                plos
                plosone
                PLoS ONE
                Public Library of Science (San Francisco, CA USA )
                1932-6203
                8 June 2017
                2017
                : 12
                : 6
                : e0179243
                Affiliations
                [1 ]College of Plant Protection, Nanjing Agricultural University/Key Laboratory of Integrated Management of Crop Diseases and Pests (Nanjing Agricultural University), Ministry of Education, Nanjing, China
                [2 ]State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing, China
                USDA-ARS Beltsville Agricultural Research Center, UNITED STATES
                Author notes

                Competing Interests: The authors have declared that no competing interests exist.

                • Conceptualization: SD YL GW.

                • Data curation: HC YL.

                • Formal analysis: HC DA YL.

                • Funding acquisition: SD YL GW.

                • Investigation: HC DA JZ YL.

                • Methodology: HC SD YL GW.

                • Project administration: SD YL GW.

                • Resources: HC DA JZ.

                • Software: HC YL.

                • Supervision: SD YL GW.

                • Validation: SD YL GW.

                • Visualization: HC DA.

                • Writing – original draft: HC SD YL GW.

                • Writing – review & editing: HC SD YL GW.

                Author information
                http://orcid.org/0000-0002-3424-7111
                Article
                PONE-D-17-00453
                10.1371/journal.pone.0179243
                5464669
                28594956
                7f1616dc-90d3-4aa0-96d2-5ffe7d62ebe0
                © 2017 Chang et al

                This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

                History
                : 4 January 2017
                : 28 May 2017
                Page count
                Figures: 5, Tables: 3, Pages: 19
                Funding
                Funded by: funder-id http://dx.doi.org/10.13039/501100001809, National Natural Science Foundation of China;
                Award ID: 31372264
                Award Recipient :
                Funded by: funder-id http://dx.doi.org/10.13039/501100001809, National Natural Science Foundation of China;
                Award ID: 31230062
                Award Recipient :
                Funded by: funder-id http://dx.doi.org/10.13039/501100001809, National Natural Science Foundation of China;
                Award ID: 31321004
                Award Recipient :
                Funded by: funder-id http://dx.doi.org/10.13039/501100001809, National Natural Science Foundation of China;
                Award ID: 31471833
                Award Recipient :
                This work was supported by National Natural Science Foundation of China (31372264 to SD, 31230062 to GW, 31321004 to GW&YL, and 31471833 to YL). The funder had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
                Categories
                Research Article
                Biology and Life Sciences
                Biochemistry
                Proteins
                Odorant Binding Proteins
                Biology and Life Sciences
                Zoology
                Animal Anatomy
                Antennae (Animal Physiology)
                Biology and Life Sciences
                Zoology
                Animal Physiology
                Antennae (Animal Physiology)
                Biology and Life Sciences
                Developmental Biology
                Life Cycles
                Larvae
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                Arthropoda
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