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1、©2012.PublishedbyTheCompanyofBiologistsLtd.1Roleoflow-densitylipoproteinreceptorrelatedprotein1(LRP1)in2CCN2/connectivetissuegrowthfactor(CTGF)proteintransportinchondrocytes311124KazumiKawata,SatoshiKubota,TakanoriEguchi,ErikoAoyama,NorifumiH.3111,*5Moritani,Se
2、ijiKondo,TakashiNishidaandMasaharuTakigawa617DepartmentofBiochemistryandMolecularDentistry,OkayamaUniversityGraduate8SchoolofMedicine,DentistryandPharmaceuticalSciences,2-5-1Shikata-Cho,9Kita-ku,Okayama-City,Okayama700-8525,Japan210BiodentalResearchCenter,Okaya
3、maUniversityDentalSchool,2-5-1Shikata-Cho,11Kita-ku,Okayama-City,Okayama700-8525,Japan312DepartmentofOralandMaxillofacialReconstructiveSurgery,OkayamaUniversity13GraduateSchoolofMedicine,DentistryandPharmaceuticalSciences,2-5-114Shikata-Cho,Kita-ku,Okayama-City
4、,Okayama700-8525,Japan1516*Authorforcorrespondence.(e-mail:takigawa@md.okayama-u.ac.jp)17JournalofCellScienceAcceptedmanuscript1JCSonlinepublicationdate27March20121Summary2LRP1isknowntobeareceptorforsignaltransmissionandendocytosis.Weformerly3reportedthatLRP1re
5、gulatesWNT/β-cateninandproteinkinaseCsignalingin4chondrocytesandrepressesthehypertrophyofchondrocytesduringendochondral5ossification,andthatLRP1isco-localizedwithaligand,CCN2,whichconducts6endochondralossification,onchondrocytes.However,theroleofLRP1inendocytot
6、ic7transportofCCN2inchondrocytesisnotyetunderstood.Inthepresentstudy,we8investigatedtheinteractionbetweenLRP1andCCN2duringendocytotictrafficking.9RNAi-mediatedknockdownofLRP1inchondrocyticHCS-2/8cellsshowedthatthe10amountofexogenousCCN2binding/incorporationwasd
7、ecreasedintheLRP111down-regulatedcells.Importantly,weobservedthatCCN2internalizationin12chondrocyteswasdependentonclathrinandinternalizatedCCN2wasco-localizedwith13anearlyorrecyclingendosomemarker.TranscytosisofCCN2throughHCS-2/8cells14wasconfirmedbyperforminge
8、xperimentswithatrans-wellapparatus,andtheamount15oftranscytosedCCN2wasdecreasedbyanLRP1antagonist.Thesefindingsruleout16possibleleakageandconfirmthecriticalinvolvementofLRP1
