A study of gentamicin injury mechanisms using cultured mouse cochlear spiral ganglion cells

(整期优先)网络出版时间:2011-01-11
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ObjectiveTostudygentamicininjurymechanismsusingpostnatalmousecochlearspiralgangcells(SGC).MethodsSGCswereisolatedusingacombinatorialapproachofenzymaticdigestionandmechanicalseparationfromP2~6Kunmingmousecochleae.After4days,culturedSGCswerefixedwith4%paraformaldehydeatroomtemperatureforimmunocytochemicalexaminationusingthemethodsofS-Pandthemonoclonalantibodyagainstmouseneurofilamentprotein(Neurofilament-68/200Kda,NF-L+H).SGCswererandomlypidedintoablankcontrolgroupandthreegentamicintreatmentgroups(mediumgentamicinconcentrationat50mg/L,100mg/Land150mg/Lrespectively),SGCswerecollectedandexaminedunderatransmissionelectronmicroscopeafterbeingculturedfor48h.ResultsSGCprimaryculturewassuccessful.SGCcytoplasmandneuritesweredyedbrownishyellowbythemonoclonalmouseneurofilamentproteinantibody.SGCsshowedclassicalbipolarneuronappearance.Underthetransmissionelectronmicroscope,.gentamicintreatedSGCsshowedmorphologicalfeaturesdifferentcomparedtothoseintheblankcontrolgroup,whichmightindicateapoptosis.ConclusionOurresultsindicatethatgentamicinhasdirecttoxiceffectsoncochlearSGCsinmiceandtheinjurymechanismiscloselyrelatedwithapoptosis.Damagetomitochondriamayplayanimportantroleintheprocess.