Estrogen Receptors: Methods and Protocols (Methods in Molecular Biology, 2418) PDF
Preview Estrogen Receptors: Methods and Protocols (Methods in Molecular Biology, 2418)
Methods in Molecular Biology 2418 Kathleen M. Eyster Editor Estrogen Receptors Methods and Protocols Second Edition M M B ETHODS IN OLECULAR IO LO GY SeriesEditor JohnM.Walker School of Lifeand MedicalSciences University ofHertfordshire Hatfield, Hertfordshire, UK Forfurther volumes: http://www.springer.com/series/7651 For over 35 years, biological scientists have come to rely on the research protocols and methodologiesinthecriticallyacclaimedMethodsinMolecularBiologyseries.Theserieswas thefirsttointroducethestep-by-stepprotocolsapproachthathasbecomethestandardinall biomedicalprotocolpublishing.Eachprotocolisprovidedinreadily-reproduciblestep-by- step fashion, opening with an introductory overview, a list of the materials and reagents neededtocompletetheexperiment,andfollowedbyadetailedprocedurethatissupported with a helpful notes section offering tips and tricks of the trade as well as troubleshooting advice. These hallmark features were introduced by series editor Dr. John Walker and constitutethekeyingredientineachandeveryvolumeoftheMethodsinMolecularBiology series. Tested and trusted, comprehensive and reliable, all protocols from the series are indexedinPubMed. Estrogen Receptors Methods and Protocols Second Edition Edited by Kathleen M. Eyster Sanford School of Medicine, University of South Dakota, Vermillion, SD, USA Editor KathleenM.Eyster SanfordSchoolofMedicine UniversityofSouthDakota Vermillion,SD,USA ISSN1064-3745 ISSN1940-6029 (electronic) MethodsinMolecularBiology ISBN978-1-0716-1919-3 ISBN978-1-0716-1920-9 (eBook) https://doi.org/10.1007/978-1-0716-1920-9 ©TheEditor(s)(ifapplicable)andTheAuthor(s),underexclusivelicensetoSpringerScience+BusinessMedia,LLC,part ofSpringerNature2022 Thisworkissubjecttocopyright.AllrightsaresolelyandexclusivelylicensedbythePublisher,whetherthewholeorpart of the material is concerned, specifically the rights of translation, reprinting, reuse of illustrations, recitation, broadcasting,reproductionon microfilmsorinanyotherphysicalway,andtransmissionorinformation storageand retrieval,electronicadaptation, computersoftware,orbysimilar ordissimilar methodologynow knownorhereafter developed. Theuseofgeneraldescriptivenames,registerednames,trademarks,servicemarks,etc.inthispublicationdoesnotimply, evenintheabsenceofaspecificstatement,thatsuchnamesareexemptfromtherelevantprotectivelawsandregulations andthereforefreeforgeneraluse. Thepublisher,theauthorsandtheeditorsaresafetoassumethattheadviceandinformationinthisbookarebelievedto betrueandaccurateatthedateofpublication.Neitherthepublishernortheauthorsortheeditorsgiveawarranty, expressedorimplied,withrespecttothematerialcontainedhereinorforanyerrorsoromissionsthatmayhavebeen made.Thepublisherremainsneutralwithregardtojurisdictionalclaimsinpublishedmapsandinstitutionalaffiliations. Coverillustration:SeeChapter6,Figure2formoredetails. ThisHumanaimprintispublishedbytheregisteredcompanySpringerScience+BusinessMedia,LLC,partofSpringer Nature. Theregisteredcompanyaddressis:1NewYorkPlaza,NewYork,NY10004,U.S.A. Preface Estrogen exerts its effects on target cells through two primary types of receptors: the genomic nuclear estrogen receptors which act as ligand-activated transcription factors, estrogen receptor α (ERα) and estrogen receptor β (ERβ), and the nongenomic membrane-bound G-protein-coupled estrogen receptor (GPER/GPR30). Traditionally, estrogenwasbestknownandmostcarefullyexaminedforitsroleinthefemalereproductive system. However, we now know that estrogen is critically important in the physiological regulationoftissuessuchasbone,brain,thecardiovascularsystem,skin,liver,adiposetissue, theimmunesystem,salivaryglands,colon,malereproductivetissues,andothers.Estrogen isafascinatinghormone,notonlybecauseofitsmultiplemechanismsofaffectingcells,that is,itsmanyreceptortypes,andforitsmanytargetorgansandcelltypes,butalsobecauseof its pleiotropic effects in health and disease. For example, a healthy vasculature responds positively to the presence of estrogen, but estrogen exacerbates the damage in a damaged vasculature. Physiologically, estrogen stimulates cell division in the breast and uterine endometrium. However, when these cell types become cancerous, the ability of estrogen to stimulate cell division is usurped to stimulate growth of the tumor. All of these physio- logicalandpathologicaleffectsofestrogenaremediatedbythesametypesofreceptorslisted above:ERα,ERβ,andGPER/GPR30. This volume includes chapters on a wide array of methods and protocols that are important to advancing our understanding of the receptor-mediated actions of estrogen. The protocols range from standard methods and important laboratory workhorses such as immunoblot and RT-qPCR to newer technologies such as PET imaging, reporter animal models,polysomefractionation,andmanyothers.Wehaveincludedprotocolsfromabroad range of tissue types to demonstrate the variety of estrogen receptor effects. We hope that scientistswhoareintriguedbythemanyfacesofestrogenwillfindtheprotocolsdetailedin thisbookhelpfulandinspirational.Thewiderangeofcutting-edgetechnologiesdescribed in this volume testifies to the continued vibrancy and excitement of the field of estrogen receptorsandestrogenactions. Iwanttoexpressmyappreciationtoalloftheauthorswhowrotechaptersforthebook and who not only tolerated my requests for further explanation on the fine points of their protocols but also responded expeditiously. My gratitude is especially profound as the authorsundertooktowritetheirchaptersamidthesignificantdisruptionstobothacademic life and personal life caused by the COVID-19 pandemic. I want to also express my appreciation to Dr. John Walker, series editor, who invited me to edit this volume and whoprovidedexcellentguidanceforthepreparationofthebook.Andtoyou,thereaders,I trust that you will find the protocols in this book useful and that the step-by-step instruc- tionswillgiveyoutheconfidencetotrynewprotocolsandexpandyourresearchrepertoire. Vermillion,SD,USA KathleenM.Eyster v Contents Preface ..................................................................... v Contributors................................................................. ix 1 AntibodyValidationforEstrogenReceptorBeta............................ 1 MadeleineBirgersson,BorbalaKatona,CeciliaLindskog, FredrikPonte´n,andCeciliaWilliams 2 ImmunoblotDetectionofthePhosphorylation oftheEstrogenReceptorαasanOutcome ofGPR30/GPER1Activation............................................ 25 SaraClark,KevinPollard,JenniferRainville, andNandiniVasudevan 3 ImmunoprecipitationAnalysesofEstrogenReceptor αPhosphorylatedatSerine216intheMouseLiver ......................... 41 MyeongJinYi,SawakoShindo,andMasahikoNegishi 4 ImmunofluorescentVerificationofSilencingEstrogenReceptor αwithsiRNAintheIntactRodentBrain .................................. 53 AnaC.Ribeiro, MihaelaA.Stavarache,SergeiMusatov, andDonaldW.Pfaff 5 DetectionandFunctionalAnalysisofEstrogenReceptor αPhosphorylatedatSerine216inMouseNeutrophils ...................... 63 SawakoShindo,RickMoore,MyeongJinYi,andMasahikoNegishi 6 QuantificationofProteinExpressionbyProximityLigation AssayintheNonhumanPrimateinResponsetoEstrogen ................... 77 ThomasW.Bonagura,JefferyS.Babischkin,GeraldJ.Pepe, andEugeneD.Albrecht 7 InSituHybridizationofEstrogenReceptorsαandβ intheHumanTestis..................................................... 95 KatjaHartmannandDanielaFietz 8 ExtractionofRNAandAnalysisofEstrogen-Responsive GenesbyRT-qPCR ..................................................... 113 MichelleL.BoozeandKathleenM.Eyster 9 PETImagingofEstrogenReceptorsUsing18F-BasedRadioligands........... 129 ManojKumar,KelleySalem,JustinJ.Jeffery,andAmyM.Fowler 10 TheUseofERE-LucReporterMicetoMonitorEstrogen ReceptorTranscriptionalActivityinaSpatio-TemporalDimension............ 153 SaraDellaTorre,ElisabettaVegeto,andPaoloCiana 11 UseofReporterGenestoAnalyzeEstrogenResponse: The7spi2;TransgenicZebrafishModel..................................... 173 DanielA.Gorelick,CarolineLucia,RuixinHao, SilviaKarim,andMariaBondesson vii viii Contents 12 AssessingEstrogenicActivityofClassicalEstrogen Receptor-BindingCompounds ........................................... 187 RichardA.PepermansandEricR.Prossnitz 13 AnOptimizedChIP-SeqProtocoltoDetermineChromatin BindingofEstrogenReceptorBeta ....................................... 203 RajithaIndukuri,AnastasiosDamdimopoulos, andCeciliaWilliams 14 PolysomeFractionationforTranscriptome-Wide Studies ofmRNATranslation ................................................... 223 JohannesRistau,KathleenWatt,ChristianOertlin, andOlaLarsson 15 Anota2seqAnalysisforTranscriptome-WideStudies ofmRNATranslation ................................................... 243 ChristianOertlin,KathleenWatt,JohannesRistau, andOlaLarsson 16 Whole-GenomeGenotypingUsingDNAMicroarrays forPopulationGenetics ................................................. 269 AustinJ.VanAsseltandErikA.Ehli 17 ProteomicsAnalysisoftheEstrogenEffectsintheRat UterusUsingGel-LCandTandemMassSpectrometryApproaches........... 289 MariaD.PaezandEduardoA.Callegari 18 ExpressionProfilesofEstrogen-RegulatedMicroRNAsinCancerCells ....... 313 AmenaArcher,ClaudiaKutter,andCeciliaWilliams 19 MolecularCloningandPurificationoftheProteinLysine MethyltransferaseSMYD2anditsCo-crystallizationwith aTargetPeptidefromEstrogenReceptorAlpha............................ 345 YingxueZhangandZheYang 20 MolecularDesign,Synthesis,andEvaluationofSNIPER(ER) thatInducesTargetedProteinDegradationofERα ......................... 363 NobumichiOhoka,HidetomoYokoo,KeiichiroOkuhira, YosukeDemizu,andMikihikoNaito 21 AssessmentofMetabolicRegulationbyEstrogenReceptors ................. 383 GeetanjaliSharmaandEricR.Prossnitz 22 RNA-SeqExperimentandDataAnalysis................................... 405 MiyurajHarishchandraHikkaduwaWithanage, HanquanLiang,andErliangZeng Index ...................................................................... 425 Contributors EUGENED.ALBRECHT • DepartmentofObstetricsandGynecology,ReproductiveSciences, UniversityofMarylandSchoolofMedicine,Baltimore,MD,USA AMENA ARCHER • SciLifeLab,DepartmentofProteinScience,KTH—RoyalInstituteof Technology,Solna,Sweden;DepartmentofBiosciencesandNutrition,Karolinska Institutet,Huddinge,Sweden JEFFERYS.BABISCHKIN • DepartmentofObstetricsandGynecology,ReproductiveSciences, UniversityofMarylandSchoolofMedicine,Baltimore,MD,USA MADELEINEBIRGERSSON • SciLifeLab,DepartmentofProteinScience,KTH—RoyalInstitute ofTechnology,Solna,Sweden;DepartmentofBiosciencesandNutrition,Karolinska Institutet,Huddinge,Sweden THOMASW.BONAGURA • DepartmentofBiology,BuenaVistaUniversity,StormLake,IA, USA MARIABONDESSON • DepartmentofIntelligentSystemsEngineering,IndianaUniversity, Bloomington,IN,USA MICHELLEL.BOOZE • SanfordFertilityandReproductiveMedicine,SiouxFalls,SD,USA EDUARDOA.CALLEGARI • DivisionofBasicBiomedicalSciences,SanfordSchoolofMedicine oftheUniversityofSouthDakota,Vermillion,SD,USA PAOLOCIANA • DepartmentofHealthSciences,UniversityofMilan,Milan,Italy SARACLARK • DepartmentofCellandMolecularBiology,TulaneUniversity,NewOrleans, LA,USA ANASTASIOS DAMDIMOPOULOS • Bioinformatics andExpressionCore,Departmentof BiosciencesandNutrition,KarolinskaInstitutet,Huddinge,Sweden SARADELLATORRE • DepartmentofPharmaceuticalSciences,UniversityofMilan,Milan, Italy YOSUKEDEMIZU • DivisionofOrganicChemistry,NationalInstituteofHealthSciences, Kawasaki,Kanagawa,Japan ERIK A.EHLI • AveraInstituteforHumanGenetics,AveraMcKennanHospitaland UniversityHealthCenter,SiouxFalls,SD,USA;DepartmentofPsychiatry,SanfordSchool ofMedicine,UniversityofSouthDakota,SiouxFalls,SD,USA KATHLEENM.EYSTER • DivisionofBasicBiomedicalSciences,SanfordSchoolofMedicine, UniversityofSouthDakota,Vermillion,SD,USA DANIELAFIETZ • InstituteforVeterinaryAnatomy,HistologyandEmbryology,JustusLiebig University,Giessen,Germany AMYM.FOWLER • DepartmentofRadiology,UniversityofWisconsinSchoolofMedicineand PublicHealth,Madison,WI,USA;UniversityofWisconsinCarboneCancerCenter, Madison,WI,USA;DepartmentofMedicalPhysics,UniversityofWisconsinSchoolof MedicineandPublicHealth,Madison,WI,USA DANIELA.GORELICK • Center forPrecisionEnvironmentalHealth,BaylorCollegeof Medicine,Houston,TX,USA RUIXIN HAO • DepartmentofBiologyandBiochemistry,Center forNuclearReceptorsand CellSignaling,UniversityofHouston,Houston,TX,USA KATJAHARTMANN • InstituteforVeterinaryAnatomy,HistologyandEmbryology,Justus LiebigUniversity,Giessen,Germany ix x Contributors MIYURAJHARISHCHANDRAHIKKADUWAWITHANAGE • DivisionofBiostatisticsand ComputationalBiology,CollegeofDentistry,UniversityofIowa,IowaCity,IA,USA RAJITHAINDUKURI • SciLifeLab,DepartmentofProteinScience,KTH—RoyalInstituteof Technology,Solna,Sweden;DepartmentofBiosciencesandNutrition,Karolinska Institutet,Huddinge,Sweden JUSTINJ.JEFFERY • UniversityofWisconsinCarboneCancerCenter,Madison,WI,USA SILVIAKARIM • DepartmentofIntelligentSystemsEngineering,IndianaUniversity, Bloomington,IN,USA BORBALAKATONA • DepartmentofImmunology,GeneticsandPathology,Uppsala University,Uppsala,Sweden MANOJKUMAR • DepartmentofRadiology,UniversityofWisconsinSchoolofMedicineand PublicHealth,Madison,WI,USA;DepartmentofRadiology,CanaryCenteratStanford forCancerEarlyDetection,StanfordSchoolofMedicine,PaloAlto,CA,USA CLAUDIAKUTTER • SciLifeLab,DepartmentofMicrobiology,TumorandCellBiology, KarolinskaInstitutet,Solna,Sweden OLALARSSON • DepartmentofOncology-Pathology,ScienceforLifeLaboratory,Karolinska Institute,Stockholm,Sweden HANQUANLIANG • McDermottCenter forHumanGrowthandDevelopment,Universityof TexasSouthwesternMedicalCenter,Dallas,TX,USA CECILIALINDSKOG • DepartmentofImmunology,GeneticsandPathology,Uppsala University,Uppsala,Sweden CAROLINELUCIA • DepartmentofBiologyandBiochemistry,Center forNuclearReceptors andCellSignaling,UniversityofHouston,Houston,TX,USA RICKMOORE • PharmacogeneticsSection,ReproductiveandDevelopmentalBiology Laboratory,NationalInstituteofEnvironmentalHealthSciences,NationalInstitutesof Health,Durham,NC,USA SERGEIMUSATOV • LaboratoryofMolecularNeurosurgery,WeillMedicalCollegeofCornell University,NewYork,NY,USA MIKIHIKONAITO • DivisionofMolecularTargetandGeneTherapyProducts,National InstituteofHealthSciences,Kawasaki,Kanagawa,Japan;DivisionofOrganicChemistry, NationalInstituteofHealthSciences,Kawasaki,Kanagawa,Japan;Laboratoryof TargetedProteinDegradation,GraduateSchoolofPharmaceuticalSciences,The UniversityofTokyo,Tokyo,Japan MASAHIKO NEGISHI • PharmacogeneticsSection,ReproductiveandDevelopmentalBiology Laboratory,NationalInstituteofEnvironmentalHealthSciences,NationalInstitutesof Health,Durham,NC,USA CHRISTIAN OERTLIN • DepartmentofOncology-Pathology,ScienceforLifeLaboratory, KarolinskaInstitute,Stockholm,Sweden NOBUMICHIOHOKA • DivisionofMolecularTargetandGeneTherapyProducts,National InstituteofHealthSciences,Kawasaki,Kanagawa,Japan KEIICHIROOKUHIRA • DivisionofMolecularTargetandGeneTherapyProducts,National InstituteofHealthSciences,Kawasaki,Kanagawa,Japan;DepartmentofEnvironment andHealthSciences,OsakaUniversityofPharmaceuticalSciences,Osaka,Japan MARIAD.PAEZ • DivisionofBasicBiomedicalSciences,SanfordSchoolofMedicineofthe UniversityofSouthDakota,Vermillion,SD,USA GERALD J.PEPE • DepartmentofPhysiologicalSciences,EasternVirginiaMedicalSchool, Norfolk,VA,USA
The list of books you might like
