Functional Hair Cell Mechanotransducer Channels Are Required for Aminoglycoside Ototoxicity 英文参考文献.docVIP

FunctionalHairCellMechanotransducerChannelsAre RequiredforAminoglycosideOtotoxicity AbdelrahmanAlharazneh1,2.,LaurenLuk1.,MarkusHuth1,AshkanMonfared1,PeterS.Steyger3,AlanG. Cheng1*,AnthonyJ.Ricci1,4* 1Department ofOtolaryngology-Head and Neck Surgery, Stanford University, Stanford, California, United States ofAmerica, 2Department ofSpecial Surgery, Mu’tah University,Alkarak,Jordan,3DepartmentofOtolaryngology-HeadandNeckSurgery,OregonHealthScienceUniversity,Portland,Oregon,UnitedStatesofAmerica, 4DepartmentofMolecularandCellularPhysiology,StanfordUniversity,Stanford,California,UnitedStatesofAmerica Abstract Aminoglycosides (AG) are commonly prescribed antibiotics with potent bactericidal activities. One main side effect is permanent sensorineural hearingloss,inducedbyselective innerearsensoryhaircelldeath.Muchworkhasfocusedon AG’s initiating cell death processes, however, fewer studies exist defining mechanisms of AG uptake by hair cells. The currentstudyinvestigatedtwoproposedmechanismsofAGtransportinmammalianhaircells:mechanotransducer (MET) channels and endocytosis. To study these two mechanisms, rat cochlear explants were cultured as whole organs in gentamicin-containing media.Two-photonimagingofTexasRedconjugatedgentamicin(GTTR)uptakeintolivehaircells was rapid and selective. Hypocalcemia, which increases the open probability of MET channels, increased AG entry into haircells.ThreeblockersofMETchannels(curare,quinine,andamiloride)significantlyreducedGTTRuptake,whereasthe endocytosis inhibitor concanavalin A did not. Dynosore quenched the fluorescence of GTTR and could not be tested. PharmacologicblockadeofMETchannelswithcurareorquinine,butnotconcanavalinAordynosore,preventedhaircell loss when challenged with gentamicin for up to 96hours. Taken together, data indicate that the patency of MET channelsmediatedAGentryintohaircellsanditstoxicity.ResultssuggestthatlimitingpermeationofAGsthroughMET channel or preventing their entry into endolymph are potential