如何国际权威杂志上发表科学论文.ppt

许田教授 HowtoGiveOralPresentations 11月9日 星期日 11月3日 星期一 顾玉东院士 怎样做临床科研 11月10日 星期一 袁钧瑛教授 科研与合作 8 50 306教室 邯郸校区正门内8 00有车前来韩启德院士 漫谈研究生的成长问题 1 30pm 12 30 教室 新视野系列研究生课程 科技论文的构思 撰写和发表 生物医学世纪讲坛 HowtoPublishYourPapersintheTopScientificJournals 鲁白教授 鲁白教授 1957年12月生于上海市 1982年获上海华东师范大学学士学位 1990年获康乃尔大学博士学位后 在洛克菲勒大学和哥伦比亚大学从事博士后研究 导师为PaulGreengard和蒲慕明教授 1993年加入罗氏分子生物学研究所 并任哥伦比亚大学助理教授 1996年加入NIH儿童健康和人类发育研究所 任突触发育和可塑性研究室主任 主要研究神经营养因子在突触发育和可塑性中的作用 他的研究室是最早提出并发现神经营养因子对神经系统突触传递 突触发育可塑性有调控作用的实验室之一 与几个著名实验室一起开创了神经营养因子的突触调控这一新领域 M F Egan M Kojima J H Callicott T E Goldberg B S Kolachana E Zaistev A Bertolino B Gold D Goldman M Dean B Lu co correspondingauthor andD R Weinberger 2003 TheBDNFval66metpolymorphismaffectsactivity dependentsecretionofBDNFandhumanmemoryandhippocampalfunction Cell112 257 269 L Ma Y Z Huang J Valtschanoff L Feng B Lu W Xiong R Weinberg L Mei 2003 Ligand dependentrecruitmentoftheneuregulinsignalingcomplexintoneuronallipidrafts J Neurosci 23 3164 3175 J Wang C Q Chen B Lu andC P Wu 2003 GDNFacutelypotentiatesCa2 channelsandexcitatorysynaptictransmissioninmidbraindopaminergicneurons NeuroSignals12 78 88 M Miura S Gronthos M Zhao B Lu L W Fisher P G Robey andS Shi 2003 SHED Stemcellsfromhumanexfoliateddeciduousteeth Proc Natl Acad Sci USA100 5807 5812 Y X Zhou M Zhao K Shimazu K Sakata D Li C X Deng B Lu 2003 ImpairmentsincerebellumPurkinjecellsandmotorfunctioninmicelackingSmad4inthecentralnervoussystem J Biol Chem 278 42313 42320 J Du L Feng E Zaitsev H S Je X Liu andB Lu 2003 Activity andtyrosinekinase dependentfacilitationofTrkBreceptorinternalizationinhippocampalneurons JCellBiol Inpress Z G Luo H S Je F Yang W C Xiong B Lu andL Mei 2003 ActivationofgeranylgenanyltransferaseisessentialforAgrin inducedAChRclustering NeuronInpress S X Bamji K Shimazu N Kimes J Huelsken W Birchmeier B Lu L F Reichardt 2003 Regulationofpresynapticassemblyandmaintenanceby catenin NeuronInPress F Yang X He J Russell andB Lu 2003 Ca2 influxindependenttransmitterreleasemediatedbymitochondrialNa Ca2 exchangerandproteinkinaseC J CellBiol Inpress 2003researcharticles WhatIsthePurposeofDoingResearch ItisnotaboutthenumberofpapersItisnotabouttheimpactfactorofthepapersItisnotabouttheNobelPrize Publishorperish1Nature 10JBC BasicElementsforBasicResearch Passion 6Sigma Silverbullettest GoodResearchIstheKey MyEnglishisnotgoodTheyarebiasedagainstChinese foreigners WhatIsaFirst ClassPaper Research Majoradvanceinaclassicfield干细胞是如何分化成特定组织细胞的 胆固醇在人体的正常功用Newtechniquesandmethodsthatcanbewidelyused人类基因组研究中的自动测序技术 PCR Patchclamp Ca2 Imaging GFPDiscoverieswithobviouspracticalimplicationsAIDSvirusreceptor的发现 老年痴呆症基因的发现Conceptualbreakthrough novelideas神经营养因子可以促进学习记忆 RNA干扰现象Challengetotraditionalviews breakdogma脑内有可分裂的神经干细胞 打破了传统观念Openingupnewarea crossboard 细胞凋亡 现象的发现 开辟了新的科研领域 WhatIsaMediocrePaper Research HorizontalgrowthImadethediscoveryinrats youfindthesameincat FillinggapsEGFactivatesJNKwhichisknowntoinducec Junexpression YoushowthatEFGenhancesc Junexpression WorkingoutdetailsIfoundNOinducestheproductionofcGMP youworkoutdoseresponseandtimecourse Supportexistingidea metoo EGF Rendocytosisrequiresdynamin PDGF Rtoo FollowupCREBbindstoCRE WorkingoutCREsequence Incompletestudy preliminary HowtoReadScientificPapers TheGilbertwayKeeptheseinmindwhenyoureadWhatisthemajorquestionaddressedinthispaper Isthisquestionimportantandwhy Whataretheapproachesusedinthispaper andwhethertheyareadequateforthequestions Whatarethenovelideaorusinginnovativeapproaches Whatistheconceptcomingoutofthispaper Dotheresultspresentedsupportthisnewconcept WeeklyreadingofCNStitlesCritical appreciative WhatMakesGoodScience ImportantandsignificantOriginalandinnovativeSolidandrigorousUniqueandunusual Noveltyisessential 语不惊人誓不休 Theevaluationprocess EditorialstaffBoardofReviewingEditors 20 30 REJECT REVIEW REJECT 70 ACCEPT 10 70 20 6 4 ShouldyourpapergotoCNS Isityourbestever Willithaveabigimpact Doesitinterestscientistsinotherfields Doesitoverturnconventionalwisdom Workthatrepresentsalargestepforwardsolutiontolong standingproblemdifferentwayofthinkingbroadimplications Whathelps ConvincingdataAppropriatecontrolsCarefulpresentationConsiderationofallviablealternatives Whatdoesn thelp Theminimalpublishableunit ExcessiveorunfoundedspeculationRepeatexamplesofaknownphenomenonInsufficientadvanceoverpreviouslypublishedwork EditorialPoliciesofDifferentJournals Cell Neuron ImmunityEditorialboarddoesalotofreviews EditorsdiscussanddecideNaturesisterjournalsEditorsdiscussanddecideScienceSpacemeeting boardofrevieweditorsPNASCommunicate contribute TrackCOthers ProceduresforHighProfileJournals Pre submissioninquirySubmit coverletterInitialscreenSendoutforreviewsReject softreject reviseRebuttalReviseagainAccept significance importancegeneralinterestsunusual surprise Initialscreening suggestreviewers maytakeonefriendsmaynotalwayssupportyou nottoreview alwayshonored soft and harsh reviewers Selectionofreviewers You Editors CoverLetters mainfindingssignificancesuggestedreviewers nottoreview listwhohaveread DearEditor Wewouldliketosubmittheenclosedmanuscriptentitled GDNFAcutelyModulatesNeuronalExcitabilityandA typePotassiumChannelsinMidbrainDopaminergicNeurons whichwewishtobeconsideredforpublicationinNatureNeuroscience GDNFhaslongbeenthoughttobeapotentneurotrophicfactorforthesurvivalofmidbraindopaminergicneurons whicharedegeneratedinParkinson sdisease Inthispaper wereportanunexpected acuteeffectofGDNFonA typepotassiumchannels leadingtoapotentiationofneuronalexcitability inthedopaminergicneuronsincultureaswellasinadultbrainslices Further weshowthatGDNFregulatestheK channelsthroughamechanismthatinvolvesactivationofMAPkinase Thus thisstudyhasrevealed forthefirsttime anacutemodulationofionchannelsbyGDNF OurfindingschallengetheclassicviewofGDNFasalong termsurvivalfactorformidbraindopaminergicneurons andsuggestthatthenormalfunctionofGDNFistoregulateneuronalexcitability andconsequentlydopaminerelease TheseresultsmayalsohaveimplicationsinthetreatmentofParkinson sdisease Duetoadirectcompetitionandconflictofinterest werequestthatDrs XXXofHarvardUniv andYYofYaleUniv notbeconsideredasreviewers Withthanksforyourconsideration IamSincerelyyours DearEditor Wewouldliketosubmittheenclosedmanuscriptentitled Ca2 bindingproteinfrequeninmediatesGDNF inducedpotentiationofCa2 channelsandtransmitterrelease whichwewishtobeconsideredforpublicationinNeuron WebelievethattwoaspectsofthismanuscriptwillmakeitinterestingtogeneralreadersofNeuron First wereportthatGDNFhasalong termregulatoryeffectonneurotransmitterreleaseattheneuromuscularsynapses Thisprovidesthefirstphysiologicalevidenceforaroleofthisnewfamilyofneurotrophicfactorsinfunctionalsynaptictransmission Second weshowthattheGDNFeffectismediatedbyenhancingtheexpressionoftheCa2 bindingproteinfrequenin Further GDNFandfrequeninfacilitatesynaptictransmissionbyenhancingCa2 channelactivity leadingtoanenhancementofCa2 influx Thus thisstudyhasidentified forthefirsttime amoleculartargetthatmediatesthelong term synapticactionofaneurotrophicfactor Ourfindingsmayalsohavegeneralimplicationsinthecellbiologyofneurotransmitterrelease DearEditor Enclosedarecopiesofamanuscriptentitled BDNFandNT 4 5PromotetheDevelopmentofLong TermPotentiationintheHippocampus whichwewishtobeconsideredforpublicationinNature Asyouknow thereisagreatdealofinterestandexcitementrecentlyinunderstandingtheroleofneurotrophinsinsynapsedevelopmentandplasticity Ourmanuscriptprovides forthefirsttime thephysiologicalevidencethatneurotrophinsregulatelong termpotentiation LTP ThemainpointofthepaperisthattheneurotrophinsBDNFandNT 4induceanearlierappearanceofLTPindevelopinghippocampus IncontrasttorecentSciencearticlebyXX sgroup we andseveralotherLTPgroups didnotseethatBDNFenhancebasalsynaptictransmissioninadullthippocampus However wefoundthatinadulthippocampus inhibitionofBDNF TrkBactivityattenuatedLTP andweaktetanusthatnormallycannotinduceLTPproducedenduringLTP Thesefindingsmayhaveimplicationsinthebasicmechanismforregulationofsynapsedevelopmentandlong termmodulationofsynapticefficacy Becauseoftherathercompetitivenatureofthefieldandtheimportantimplicationofourfindings wehavenotyetpresentedthisworkinanypublicforum However confidentialdiscussionwithseveralprominentneuroscientistssuchas111and222havegeneratedtremendousexcitement Thus wefeelthatthisworkisofgeneralinterestandissuitableforpublicationinNature WewouldliketosuggestDrs aaaofYaleUniv bbbofHarvardMedicalSchool andcccofUniv ofCalifornia Berkeley asreviewersforthismanuscript Duetoadirectcompetitionandconflictofinterest werequestthatDr XXandYY notbeconsideredasreviewers Thankyouverymuchforyourconsideration Titles Important significantUnexpected unusualFunctionMechanismsSimpleStraightforwardSpecific Structure mechanism anregulationoftheNeurosporaplasmamembraneH ModulationofpostendocyticsortingofG protein coupledreceptors DistinctmolecularmechanismforinitiatingTRAF6signaling Identificationof Roleof Involvementof Sequenceofwriting AbstractFigurelayoutFigurelegendMaterialandmethodsResultsIntroductionDiscussion Abstract Rationale remainunknown Todetermine Summarystatement Hereweshow BodyDon tgointodetails don tusemanyspecialtermsSignificanceMustpointout butdon tclaimtoomuch FormationofthenormalmammaliancerebralcortexrequiresthemigrationofGABAergicinhibitoryinterneuronsfromanextracorticalorigin thelateralganglioniceminence LGE Mechanismsguidingthemigratorydirectionoftheseneurons orotherneuronsintheneocortex arenotwellunderstood WehaveusedanexplantassaytostudyGABAergicneuronalmigrationandfoundthattheventricularzone VZ oftheLGEisrepulsivetoGABAergicneurons Furthermore thesecretedproteinSlitisachemorepellentguidingthemigratorydirectionofGABAergicneurons andblockadeofendogenousSlitsignalinginhibitstherepulsiveactivityintheVZ TheseresultshaverevealedacellularsourceofguidanceforGABAergicneurons demonstratedamolecularcueimportantforcorticaldevelopment andsuggestedaguidancemechanismforthemigrationofextracorticalneuronsintotheneocortex Ithasnotescapedournoticethatthespecificpairingwehavepostulatedimmediatelysuggestsapossiblecopyingmechanismforthegeneticmaterial J D WatsonandF H C Crick Neuronalresponsestobrain derivedneurotrophicfactor BDNF areinitiatedbytheactivationofthereceptorTrkBtyrosinekinase 1 Inthisstudyweexaminedwhethercholesterol andglycolipid richmicrodomains lipidrafts provideafunctionalplatformforBDNF dependentsignaltransduction 2 Usingprimarycultureofcorticalneurons wedemonstratedthatTrkBwasdramaticallytranslocatedintolipidraftsinBDNF dependentmanner 3 ThistranslocationwasblockedbythepharmacologicaleffectofgeneralTrkinhibitors indicatingthatTrkBactivationisrequiredforthetranslocationmechanism WealsoshowedthatBDNFandTrkB FLwerebothconcentratedinlipidraftsduringdevelopmentofcerebralcortex concomitantwiththatofsynapticvesicleproteins includingsolubleN ethylmaleimide sensitivefactorattachmentproteinreceptor SNARE proteinsandsynaptophysin 4 Thisresult togetherwiththefindingsthatBDNFstimulationcausedtranslocationofsynaptophysinintolipidrafts 5 andthatBDNF enhancedglutamatereleaseandexocytosiswerebothattenuatedbydepletionofcholesterolfromthecellsurfacewithmethyl beta cyclodextrin MCD indicatesthatlipidraftsareessentialforBDNFregulationofneurotransmitterrelease 6 Brain derivedneurotrophicfactor BDNF playsanimportantroleinsynapticplasticitybuttheunderlyingsignalingmechanismsremainunknown HereweshowthatBDNFrapidlyrecruitsfull lengthTrkB TrkB FL receptorintocholesterol richlipidraftsfromnon raftregionsofneuronalplasmamembranes TruncatedTrkBlackingtheintracellularkinasedomainwasnottranslocated andthetranslocationofTrkB FLwasblockedbyTrkinhibitors suggestingaroleforTrkBtyrosinekinaseinthetranslocation DisruptionoflipidraftsbydepletingcholesterolfromthecellsurfaceblockedBDNF dependentTrkBtranslocation DisruptionofraftsalsopreventedthepotentiatingeffectofBDNFontransmitterreleaseinculturedneurons aswellasthatonsynapticresponsetotetanusinhippocampalslices Incontrast lipidraftsarenotrequiredforBDNFregulationofneuronalsurvival Thus ligand inducedTrkBtranslocationintolipidraftsmayrepresentaselectivesignalingmechanismforsynapticmodulationbyBDNFintheCNS Acalcium independentbutvoltage dependentsecretion CIVDS coexistswiththecalciumdependentexocytosisindorsalrootganglion DRG neurons 1 HerewehaveinvestigatedtheCIVDS coupledendocytosis 2 Usingopticalandmembranecapacitancemeasurements weshowthat incalcium freemedium eitherstepdepolarizationoratrainofaction potential likestimulationinduceanovelformofrapidendocytosis whichoccursimmediatelyaftertheCIVDS Surprisingly thiscalcium independentendocytosisisstronglydependentonthestimulationfrequency 3 H7suppresstheendocytosis whilePKAagonistsenhanceit 4 Biochemicalexperimentsshowthatmembranedepolarizationdirectlyup regulatePKAinDRGneurons OurexperimentsalsoshowedthatthefrequencydependencyofCIVDS REisdynamin independent 5 Thus ourdataindicatethatneuronalactivitymodulatesarapidendocytosisviaaCa2 anddynamin independentphosphorylation dependentmannerinDRGneurons 6 SynapticvesicleendocytosisisbelievedtorequireCa2 andtheGTPasedynamin Herewereportanovelformofrapidendocytosis RE thatisindependentofCa2 anddynaminindorsalrootganglion DRG neurons UsingFMdyelabelingandmembranecapacitancemeasurements weshowthatbothstepdepolarizationandrepetitivestimulationinduceREinCa2 freemedium REalsooccursinthepresenceofaCa2 chelator BAPTA InhibitionofdynaminfunctionbythreedifferentapproachesdoesnotaffectRE ProteinkinaseA PKA inhibitorssuppresstheendocytosis whilePKAactivatorsenhanceit Biochemicalexperimentsdemonstratethatmembranedepolarizationdirectlyup regulatedPKAactivity TheseresultsrevealaCa2 anddynamin independentformofendocytosisthatiscontrolledbyneuronalactivityandPKA dependentphosphorylationinDRGneurons Introduction Whatdoweknowaboutthesubjects Onlyrelevantinformationshouldbeprovided don twriteareviewWhatwedon tknowRationaleWhyyouwanttodoit Don trepeatabstractApproachesHowyouaregoingtodoit SignificanceMakeanappealtogeneralreaders InthisstudywehaveexaminedtheroleofchromograninsCGAandCGB indense coresecretorygranulebiogenesis WeanalyzedtheeffectofspecificdepletionofeitherCGAorCGB usinganantisenseRNAstrategy ondense coresecretorygranuleformationinratpheochromocytoma PC12 cells amodelneuroendocrinecellline WealsoexpressedCGAinapituitarycellline 6T3 lackingtheregulatedsecretorypathwayandnonendocrinefibroblastcellstodetermineitseffectoninductionofdense coresecretorygranulebiogenesisandregulatedsecretion Finally wedeterminedwhetherCGAcouldregulatethelevelofothersecretorygranuleproteinsinneuroendocrineandendocrinecells PC12and6T3 ThesestudiesidentifiedCGAasakeyregulatorofdense coresecretorygranulebiogenesisandstorageofothergranuleproteinsinendocrinecells Results LogicNeedtoexplaintherationalesinthebeginningConnectionsbetweenparagraphsDon tjump Previousstudieshaveshownthatmembranedepolarization triggeredCa2 influxthroughL typeVSCCsinducesanincreaseinBDNFmRNAexpressioninculturedneurons Zafraetal 1990 Ghoshetal 1994 ThisincreaseinBDNFmRNAcouldbetheresultofincreasedtranscriptioninitiation orincreasedBDNFmRNAstability orboth TodetermineifmembranedepolarizationstimulatesBDNFtranscription we GiventhefindingthatCa2 influxthroughL typeVSCCsinducesBDNFtranscription experimentswerecarriedouttodeterminewhichofthefourBDNFpromotersiscapableofmediatingaCa2 response Asdescribedabove theratBDNFgeneconsistsoffourdistinct5 exonseachdrivenbyaspecificpromoterandeachsplicedtoacommon3 exonthatencodestheBDNFprotein SinceeachofthefourprimaryBDNFtranscriptscanbepolyadenylatedatoneoftwosites atotalofeightBDNFtranscriptsaregenerated Inprinciple theeighttranscriptscanbedistinguishedbyNorthernblottingusing5 exon specificprobes sinceeachofthefour5 exonprobesshoulddetectashortandalongBDNFtranscript ByidentifyingthespecificBDNFmRNAsinduceduponCa2 influxthroughL typeVSCCs itshouldbepossibletoidentifywhichofthefourBDNFpromotersisCa2 responsive sincetheCa2 responsivepromoter s wouldbeexpectedtobelocatedjust5 oftheinitiationsiteofBDNFmRNAsynthesis Discussion SummaryofmainfindingsPapersthatsupportyou butdon tdowngradeyournoveltyPitfallsandwhySignificance Don tspeculatetoomuch Theresultsinthepresentstudymayhaveanumberofimplicationsinthecellbiologyoftyrosinekinasereceptors First wereportthe Toourknowledge thisisthefirstdemonstrationfor Thus ourresultssuggestacross talkbetweenCa2 andtyrosinekinasesignalingpathways Second thepresentstudyrevealsanimportantregulatoryeffectof Itwillbeinterestingtodeterminewhether Finally weshowthat Xxx Takentogether theseresultssuggestageneralroleoftyrosinekinaseintheendocytosisofgrowthfactorreceptors Therearethreemainfindingsinthepresentstudy First wereportaGDNF inducedlong termfacilitationofneurotransmitterreleaseattheneuromuscularsynapses Second weshowthattheeffectofGDNFonsynaptictransmissionismediatedbyanincreaseintheexpressionoftheCa2 bindingproteinfrequenin Finally wedemonstratethatGDNFandfrequeninfacilitatesynaptictransmissionbyenhancingN typeCa2 channelactivation leadingtoanenhancementofCa2 influx Thus thisstudyhasidentified forthefirsttime amoleculartargetthatmediatesthelong term synapticactionofaneurotrophicfactor Ourfindingsmayalsoprovidenewinsightsintotheregulatorymechanismsofneurotransmitterrelease Invitethoroughcritique RunyourownreviewprocessfirstaskfeedbackfromsomeoneinyourownspecialtysomeoneinanunrelatedspecialtyagoodeditorfortheEnglishlanguageAssessbothresearchandpresentation Commonreasonsforrejection BelongsinaspecializedjournalToosmallofanadvanceoverpreviouslypublishedworkUnconvincingdataObservationswithoutinterpretationsInterpretationswithoutdata Reviseyourpaper Becalmaboutreviewerscriticisms AlwaysmakeeditoryourfriendNeverarguewithreviewersTrytodoeverythingthatreviewersaskSeizetheopportunitywhenreviewersmakemistakes Whenyourpapergetsrejected withoutreview DearEditor Iwouldappreciateifyoucouldreconsidertoreviewourmanuscript 111 Wefeelstronglythatthisisanimportantsubjectthattouchesoneofthecentraldogmasinneuroscience xxx Itisalsoverytimely giventhepublicationofthepaperbyXandYentitled 222 inthelatestissueofNatureNeuroscience Inthispaper theauth