RNAi-based demonstration of direct link between specific odorant receptors and mosquito oviposition behavior
The Southern house mosquito, Culex quinquefasciatus – a vector of West Nile virus – is equipped with 130 odorant receptors (ORs), which enable young females to locate plants and blood-meal sources and older females to find suitable sites for oviposition. In our attempts to de-orphanize ORs expressed...
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description | The Southern house mosquito, Culex quinquefasciatus – a vector of West Nile virus – is equipped with 130 odorant receptors (ORs), which enable young females to locate plants and blood-meal sources and older females to find suitable sites for oviposition. In our attempts to de-orphanize ORs expressed in female antennae, we identified CquiOR37 and CquiOR99, which were narrowly tuned to two phenolic compounds, 4-methylphenol and 4-ethylphenol. When tested in the Xenopus oocyte recording system the observed EC50s for 4-methylphenol and 4-ethylphenol were 6.4 and 18.2 μM for CquiOR37 and 14.4 and 0.74 μM for CquiOR99 (goodness of fit, R2 = 0.88–0.99), respectively. Indoor behavioral assays demonstrated that gravid female mosquitoes laid significantly more eggs in water trays spiked with these compounds than in control water trays. Field studies with gravid traps corroborated that 4-ethylphenol is active in a wide range of doses from 0.1 to 10 μg/l, as required for practical applications. A dsRNA construct based on the two genes, CquiOR37/99-dsRNA was stable in pupa hemolymph for up to 3 h. Pupae injected with CquiOR37/99-dsRNA, β-galactosidase-dsRNA or water had more than 40% survival rate at the peak of oviposition (day-9). qPCR analysis showed individual variation, but significant mean reduction in CquiOR37 and CquiOR99 transcript levels in CquiOR37/99-dsRNA-treated mosquitoes. Water-injected females and those treated with the control gene laid significantly more eggs in trays containing 4-ethylphenol than in water trays, whereas CquiOR37/99-dsRNA-treated mosquitoes laid normal number of eggs, but could not discriminate treatment from control. This study linked for the first time specific receptors for 4-ethylphenol with increased oviposition in the important vector Cx. quinquefasciatus.
[Display omitted]
•Identification and characterization of Culex odorant receptors narrowly tuned to phenolic compounds.•Evaluation of oviposition attractants by indoor assays and field tests.•Silencing of odorant receptor genes and behavioral evaluation of the phenotype. |
doi_str_mv | 10.1016/j.ibmb.2013.07.008 |
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[Display omitted]
•Identification and characterization of Culex odorant receptors narrowly tuned to phenolic compounds.•Evaluation of oviposition attractants by indoor assays and field tests.•Silencing of odorant receptor genes and behavioral evaluation of the phenotype.</description><identifier>ISSN: 0965-1748</identifier><identifier>EISSN: 1879-0240</identifier><identifier>DOI: 10.1016/j.ibmb.2013.07.008</identifier><identifier>PMID: 23911547</identifier><language>eng</language><publisher>England: Elsevier Ltd</publisher><subject>4-Ethylphenol ; 4-Methylphenol ; Animals ; antennae ; Behavior, Animal ; blood meal ; Cloning, Molecular ; CquiOR37 ; CquiOR99 ; Cresols - metabolism ; Culex - metabolism ; Culex quinquefasciatus ; double-stranded RNA ; eggs ; Female ; females ; genes ; hemolymph ; Insect Vectors - metabolism ; Mosquito Control ; odor compounds ; oocytes ; Oviposition ; oviposition sites ; phenolic compounds ; Phenols - metabolism ; Pheromones - metabolism ; pupae ; quantitative polymerase chain reaction ; receptors ; Receptors, Odorant - metabolism ; RNA Interference ; RNAi ; survival rate ; traps ; trays ; West Nile virus</subject><ispartof>Insect biochemistry and molecular biology, 2013-10, Vol.43 (10), p.916-923</ispartof><rights>2013 The Authors</rights><rights>Copyright © 2013 The Authors. Published by Elsevier Ltd.. All rights reserved.</rights><rights>2013 The Authors, Published by Elsevier Ltd. All rights reserved. 2013</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c479t-be00e08a62e2c33e22723fb072d56ed22a70f5aac8a5e4630e7bc64e971e8d653</citedby><cites>FETCH-LOGICAL-c479t-be00e08a62e2c33e22723fb072d56ed22a70f5aac8a5e4630e7bc64e971e8d653</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://dx.doi.org/10.1016/j.ibmb.2013.07.008$$EHTML$$P50$$Gelsevier$$Hfree_for_read</linktohtml><link.rule.ids>230,314,780,784,885,3550,27924,27925,45995</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/23911547$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Zhu, Fen</creatorcontrib><creatorcontrib>Xu, Pingxi</creatorcontrib><creatorcontrib>Barbosa, Rosângela M.R.</creatorcontrib><creatorcontrib>Choo, Young-Moo</creatorcontrib><creatorcontrib>Leal, Walter S.</creatorcontrib><title>RNAi-based demonstration of direct link between specific odorant receptors and mosquito oviposition behavior</title><title>Insect biochemistry and molecular biology</title><addtitle>Insect Biochem Mol Biol</addtitle><description>The Southern house mosquito, Culex quinquefasciatus – a vector of West Nile virus – is equipped with 130 odorant receptors (ORs), which enable young females to locate plants and blood-meal sources and older females to find suitable sites for oviposition. In our attempts to de-orphanize ORs expressed in female antennae, we identified CquiOR37 and CquiOR99, which were narrowly tuned to two phenolic compounds, 4-methylphenol and 4-ethylphenol. When tested in the Xenopus oocyte recording system the observed EC50s for 4-methylphenol and 4-ethylphenol were 6.4 and 18.2 μM for CquiOR37 and 14.4 and 0.74 μM for CquiOR99 (goodness of fit, R2 = 0.88–0.99), respectively. Indoor behavioral assays demonstrated that gravid female mosquitoes laid significantly more eggs in water trays spiked with these compounds than in control water trays. Field studies with gravid traps corroborated that 4-ethylphenol is active in a wide range of doses from 0.1 to 10 μg/l, as required for practical applications. A dsRNA construct based on the two genes, CquiOR37/99-dsRNA was stable in pupa hemolymph for up to 3 h. Pupae injected with CquiOR37/99-dsRNA, β-galactosidase-dsRNA or water had more than 40% survival rate at the peak of oviposition (day-9). qPCR analysis showed individual variation, but significant mean reduction in CquiOR37 and CquiOR99 transcript levels in CquiOR37/99-dsRNA-treated mosquitoes. Water-injected females and those treated with the control gene laid significantly more eggs in trays containing 4-ethylphenol than in water trays, whereas CquiOR37/99-dsRNA-treated mosquitoes laid normal number of eggs, but could not discriminate treatment from control. This study linked for the first time specific receptors for 4-ethylphenol with increased oviposition in the important vector Cx. quinquefasciatus.
[Display omitted]
•Identification and characterization of Culex odorant receptors narrowly tuned to phenolic compounds.•Evaluation of oviposition attractants by indoor assays and field tests.•Silencing of odorant receptor genes and behavioral evaluation of the phenotype.</description><subject>4-Ethylphenol</subject><subject>4-Methylphenol</subject><subject>Animals</subject><subject>antennae</subject><subject>Behavior, Animal</subject><subject>blood meal</subject><subject>Cloning, Molecular</subject><subject>CquiOR37</subject><subject>CquiOR99</subject><subject>Cresols - metabolism</subject><subject>Culex - metabolism</subject><subject>Culex quinquefasciatus</subject><subject>double-stranded RNA</subject><subject>eggs</subject><subject>Female</subject><subject>females</subject><subject>genes</subject><subject>hemolymph</subject><subject>Insect Vectors - metabolism</subject><subject>Mosquito Control</subject><subject>odor compounds</subject><subject>oocytes</subject><subject>Oviposition</subject><subject>oviposition sites</subject><subject>phenolic compounds</subject><subject>Phenols - metabolism</subject><subject>Pheromones - metabolism</subject><subject>pupae</subject><subject>quantitative polymerase chain reaction</subject><subject>receptors</subject><subject>Receptors, Odorant - metabolism</subject><subject>RNA Interference</subject><subject>RNAi</subject><subject>survival rate</subject><subject>traps</subject><subject>trays</subject><subject>West Nile virus</subject><issn>0965-1748</issn><issn>1879-0240</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2013</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp9kU1v1DAQhi0EokvhD3AAH7kkjL9iR0JIVcWXVIEE9Gw59qT1ksRbO7uIf0-2KRVcOM1hnnln7IeQ5wxqBqx5va1jN3Y1ByZq0DWAeUA2zOi2Ai7hIdlA26iKaWlOyJNStgAgpdKPyQkXLWNK6g0Zvn4-i1XnCgYacExTmbObY5po6mmIGf1Mhzj9oB3OPxEnWnboYx89TSFlN810QXA3p1yomwIdU7nZxznRdIi7VOJtVIfX7hBTfkoe9W4o-OyunpLL9---n3-sLr58-HR-dlF5qdu56hAAwbiGI_dCIOeai74DzYNqMHDuNPTKOW-cQtkIQN35RmKrGZrQKHFK3q65u303YvA4LY8a7C7H0eVfNrlo_-1M8dpepYMVBkApswS8ugvI6WaPZbZjLB6HwU2Y9sUyKZSRrRLHXXxFfU6lZOzv1zCwR012a4-a7FGTBW0XTcvQi78PvB_542UBXq5A75J1VzkWe_ltSVCLQwPtLfFmJXD5yEPEbIuPOHlcpdmQ4v8u-A1v7rBt</recordid><startdate>20131001</startdate><enddate>20131001</enddate><creator>Zhu, Fen</creator><creator>Xu, Pingxi</creator><creator>Barbosa, Rosângela M.R.</creator><creator>Choo, Young-Moo</creator><creator>Leal, Walter S.</creator><general>Elsevier Ltd</general><scope>6I.</scope><scope>AAFTH</scope><scope>FBQ</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope><scope>5PM</scope></search><sort><creationdate>20131001</creationdate><title>RNAi-based demonstration of direct link between specific odorant receptors and mosquito oviposition behavior</title><author>Zhu, Fen ; Xu, Pingxi ; Barbosa, Rosângela M.R. ; Choo, Young-Moo ; Leal, Walter S.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c479t-be00e08a62e2c33e22723fb072d56ed22a70f5aac8a5e4630e7bc64e971e8d653</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2013</creationdate><topic>4-Ethylphenol</topic><topic>4-Methylphenol</topic><topic>Animals</topic><topic>antennae</topic><topic>Behavior, Animal</topic><topic>blood meal</topic><topic>Cloning, Molecular</topic><topic>CquiOR37</topic><topic>CquiOR99</topic><topic>Cresols - metabolism</topic><topic>Culex - metabolism</topic><topic>Culex quinquefasciatus</topic><topic>double-stranded RNA</topic><topic>eggs</topic><topic>Female</topic><topic>females</topic><topic>genes</topic><topic>hemolymph</topic><topic>Insect Vectors - metabolism</topic><topic>Mosquito Control</topic><topic>odor compounds</topic><topic>oocytes</topic><topic>Oviposition</topic><topic>oviposition sites</topic><topic>phenolic compounds</topic><topic>Phenols - metabolism</topic><topic>Pheromones - metabolism</topic><topic>pupae</topic><topic>quantitative polymerase chain reaction</topic><topic>receptors</topic><topic>Receptors, Odorant - metabolism</topic><topic>RNA Interference</topic><topic>RNAi</topic><topic>survival rate</topic><topic>traps</topic><topic>trays</topic><topic>West Nile virus</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Zhu, Fen</creatorcontrib><creatorcontrib>Xu, Pingxi</creatorcontrib><creatorcontrib>Barbosa, Rosângela M.R.</creatorcontrib><creatorcontrib>Choo, Young-Moo</creatorcontrib><creatorcontrib>Leal, Walter S.</creatorcontrib><collection>ScienceDirect Open Access Titles</collection><collection>Elsevier:ScienceDirect:Open Access</collection><collection>AGRIS</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>Insect biochemistry and molecular biology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Zhu, Fen</au><au>Xu, Pingxi</au><au>Barbosa, Rosângela M.R.</au><au>Choo, Young-Moo</au><au>Leal, Walter S.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>RNAi-based demonstration of direct link between specific odorant receptors and mosquito oviposition behavior</atitle><jtitle>Insect biochemistry and molecular biology</jtitle><addtitle>Insect Biochem Mol Biol</addtitle><date>2013-10-01</date><risdate>2013</risdate><volume>43</volume><issue>10</issue><spage>916</spage><epage>923</epage><pages>916-923</pages><issn>0965-1748</issn><eissn>1879-0240</eissn><abstract>The Southern house mosquito, Culex quinquefasciatus – a vector of West Nile virus – is equipped with 130 odorant receptors (ORs), which enable young females to locate plants and blood-meal sources and older females to find suitable sites for oviposition. In our attempts to de-orphanize ORs expressed in female antennae, we identified CquiOR37 and CquiOR99, which were narrowly tuned to two phenolic compounds, 4-methylphenol and 4-ethylphenol. When tested in the Xenopus oocyte recording system the observed EC50s for 4-methylphenol and 4-ethylphenol were 6.4 and 18.2 μM for CquiOR37 and 14.4 and 0.74 μM for CquiOR99 (goodness of fit, R2 = 0.88–0.99), respectively. Indoor behavioral assays demonstrated that gravid female mosquitoes laid significantly more eggs in water trays spiked with these compounds than in control water trays. Field studies with gravid traps corroborated that 4-ethylphenol is active in a wide range of doses from 0.1 to 10 μg/l, as required for practical applications. A dsRNA construct based on the two genes, CquiOR37/99-dsRNA was stable in pupa hemolymph for up to 3 h. Pupae injected with CquiOR37/99-dsRNA, β-galactosidase-dsRNA or water had more than 40% survival rate at the peak of oviposition (day-9). qPCR analysis showed individual variation, but significant mean reduction in CquiOR37 and CquiOR99 transcript levels in CquiOR37/99-dsRNA-treated mosquitoes. Water-injected females and those treated with the control gene laid significantly more eggs in trays containing 4-ethylphenol than in water trays, whereas CquiOR37/99-dsRNA-treated mosquitoes laid normal number of eggs, but could not discriminate treatment from control. This study linked for the first time specific receptors for 4-ethylphenol with increased oviposition in the important vector Cx. quinquefasciatus.
[Display omitted]
•Identification and characterization of Culex odorant receptors narrowly tuned to phenolic compounds.•Evaluation of oviposition attractants by indoor assays and field tests.•Silencing of odorant receptor genes and behavioral evaluation of the phenotype.</abstract><cop>England</cop><pub>Elsevier Ltd</pub><pmid>23911547</pmid><doi>10.1016/j.ibmb.2013.07.008</doi><tpages>8</tpages><oa>free_for_read</oa></addata></record> |
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subjects | 4-Ethylphenol 4-Methylphenol Animals antennae Behavior, Animal blood meal Cloning, Molecular CquiOR37 CquiOR99 Cresols - metabolism Culex - metabolism Culex quinquefasciatus double-stranded RNA eggs Female females genes hemolymph Insect Vectors - metabolism Mosquito Control odor compounds oocytes Oviposition oviposition sites phenolic compounds Phenols - metabolism Pheromones - metabolism pupae quantitative polymerase chain reaction receptors Receptors, Odorant - metabolism RNA Interference RNAi survival rate traps trays West Nile virus |
title | RNAi-based demonstration of direct link between specific odorant receptors and mosquito oviposition behavior |
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