Gene Targeting of the Cysteine Peptidase Cathepsin H Impairs Lung Surfactant in Mice 英文参考文献.docVIP

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Gene Targeting of the Cysteine Peptidase Cathepsin H Impairs Lung Surfactant in Mice 英文参考文献.doc

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Gene Targeting of the Cysteine Peptidase Cathepsin H Impairs Lung Surfactant in Mice 英文参考文献

GeneTargetingoftheCysteinePeptidaseCathepsinH ImpairsLungSurfactantinMice FrankBu¨hling1.,MartinKouadio2.,CarolineE.Chwieralski3.,UrsulaKern2,4,5.,JensM.Hohlfeld6, NicoleKlemm2,NicoleFriedrichs2,WeraRoth2,JanM.Deussing2¤,ChristophPeters2,7 ,Thomas Reinheckel2,7 * 1Institute of Laboratory Medicine, Carl-Thiem-Klinikum, Cottbus, Germany, 2Institut of Molecular Medicine and Cell Research, Albert-Ludwigs-University, Freiburg, Germany,3InsituteofMolecularandClinicalImmunology,Otto-von-GuerickeUniversity,Magdeburg,Germany,4FacultyofBiology,Albert-Ludwigs-University,Freiburg, Germany,5SpemannGraduateSchoolofBiologyandMedicine(SGBM),Albert-Ludwigs-University,Freiburg,Germany,6DivisionofImmunology,AllergologyandAirway Research,FraunhoferInstituteofToxicologyandExperimentalMedicine,Hannover,Germany,7BIOSSCentreofBiologicalSignallingStudies,Albert-Ludwigs-University, Freiburg,Germany Abstract Background:The11humancysteinecathepsinsareproteasesmainlylocatedintheendolysosomalcompartmentofallcells and within the exocytosis pathways of some secretory cell types. Cathepsin H (Ctsh) has amino- and endopeptidase activities.InvitrostudieshavedemonstratedCtshinvolvementintheprocessingandsecretionofthepulmonarysurfactant proteinB(SP-B).Furthermore,CtshishighlyexpressedinthesecretoryorganellesofalveolartypeIIpneumocyteswherethe surfactantproteinsareprocessed. Methodology/Principal Findings: Hence, we generated Ctsh null mice by gene targeting in embryonic stem cells to investigate the role of this protease in surfactant processing in vivo. The targeting construct contains a ?-galactosidase (lacZ) reporter enabling the visualisation of Ctsh expression sites. Ctsh-deficiency was verified by northern blot, western blot,andmeasurementoftheCtshaminopeptidaseactivity.Ctsh2/2miceshownogrossphenotypeandtheirdevelopment 2/2 is normal without growth retardation. Broncho-alveolar lavage (BAL) from Ctsh mice contained lower levels of SP-B indicatingreducedSP-Bsecretion.TheBALphospholipidconcentra