JournalofVisualizedExperimentswww.jove.comCopyright©2011CreativeCommonsAttribution-NonCommercialLicenseApril2011|50|e2732|Page1of6VideoArticleACalciumBioluminescenceAssayforFunctionalAnalysisofMosquito(Aedesaegypti)andTick(Rhipicephalusmicroplus)GProtein-coupledReceptorsHsiao-LingLu1,CymonN.Kersch1,SuparnaTaneja-Bageshwar1,2,PatriciaV.Pietrantonio11DepartmentofEntomology,TexasA&MUniversity(TAMU)2DepartmentofMolecularandCellularMedicine,TexasA&MUniversity(TAMU)Correspondenceto:PatriciaV.Pietrantonioatp-pietrantonio@tamu.eduURL:http://www.jove.com/video/2732DOI:doi:10.3791/2732Keywords:Immunology,Issue50,Aequorincalciumreporter,coelenterazine,Gprotein-coupledreceptor(GPCR),CHO-K1cells,mammaliancellculture,neuropeptideSARstudies(SAR=structure-activityrelationships),receptor-neuropeptideinteraction,bioluminescence,drugdiscovery,semi-throughputscreeninginplatesDatePublished:4/20/2011Citation:Lu,H.L.,Kersch,C.N.,Taneja-Bageshwar,S.,Pietrantonio,P.V.ACalciumBioluminescenceAssayforFunctionalAnalysisofMosquito(Aedesaegypti)andTick(Rhipicephalusmicroplus)GProtein-coupledReceptors.J.Vis.Exp.(50),e2732,doi:10.3791/2732(2011).AbstractArthropodhormonereceptorsarepotentialtargetsfornovelpesticidesastheyregulatemanyessentialphysiologicalandbehavioralprocesses.ThemajorityofthembelongtothesuperfamilyofGprotein-coupledreceptors(GPCRs).Wehavefocusedoncharacterizingarthropodkininreceptorsfromthetickandmosquito.Arthropodkininsaremultifunctionalneuropeptideswithmyotropic,diuretic,andneurotransmitterfunction.Here,amethodforsystematicanalysesofstructure-activityrelationshipsofinsectkininsontwoheterologouskininreceptor-expressingsystemsisdescribed.Weprovideimportantinformationrelevanttothedevelopmentofbiostablekininanalogswiththepotentialtodisruptthediuretic,myotropic,and/ordigestiveprocessesinticksandmosquitoes.Thekininreceptorsfromthesoutherncattletick,Boophilusmicroplus(Canestrini),andthemosquitoAedesaegypti(Linnaeus),werestablyexpressedinthemammaliancelllineCHO-K1.Functionalanalyseso...
