25722.大麻素受體對神經(jīng)干細胞分化后神經(jīng)突起生長的影響及其信號轉(zhuǎn)導機制_第1頁
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1、浙江大學碩士學位論文大麻素受體對神經(jīng)干細胞分化后神經(jīng)突起生長的影響及其信號轉(zhuǎn)導機制姓名:鐘婷婷申請學位級別:碩士專業(yè):生物化學與分子生物學指導教師:孫益201009浙江大學碩士學位論文EffectsandsignaltransductionmechanismofcannabinoidreceptorsonneuriteoutgrowthofdifferentiatedneuralstemcellsAbstractWefocusedour

2、studyontheeffectsofcannabinoidreceptoronneuriteoutgrowthofneuronscomingfrommouseneuralstemcellinvitroandthepossiblesignaltransductionmechanismWefirstlyculturedneuralstemcells(NSCs、insuspensioninserumfreemediumcontainingb

3、asicflbroblastgrowthfactor(bFGF)andepidermalgrowthfactor(EGF)111eNSCsarefloatedasmulti—cellaggregatescalledneurospheresinvitronleneurosphereswhicharenestinpositiveCanbepassagedandCandifferentiatetoneuronsandgliacellsInor

4、dertostudytheeffectsofcananbinoidreceptorsonneuriteoutgrowth,weinducedtheNSCstodifferentiatemostlyintoneUmnSTheendogenouscannabinoidsystemhasbeenfoundwidelyexpressedinvarioustissuesandorgansofmammalianTodate,twomajorcann

5、abinoidreceptorshavebeendescribed,CB1receptors(CB1R)andCB2receptors(CB2R)CB1RispresentmainlyinCNSandmodulatekindsofneurophysiologyfunctionsTheCB2RisthoughttobeexpressedonlyinperipheralsystemHoweverithasbeenreportedrecent

6、lythatCB2RisalsolocalizedindifferentcellpopulationsinCNSTodoSO,weinvestigatedthepresenceoftheCBlRandCB2RmRNAsintheneurons,astrocytes,NSCsanddifferentiatedcellsfromNSCsbyRT—PCRWefoundthatCB1RandCB2RmRNAswereexpressedbya11

7、thekindsofthesecellsSubsequently,WestudiedtheeffectsofWIN552122(WIN),amixedCB1R/CB2Ragonist,onneuriteoutgrowthofdifferentiatedcellsfromNSCsinvitroThroughtheneuriteoutgrowthassay,WINhadbeenshowedtoimproveneuriteoutgrowthi

8、naconcentration—dependentmannerTobettercharacterizetheinvolvementofeachsingleCBreceptorinWINenhancedneuriteoutgrowth,wecoincubatedthedifferentiatedcellswithWINandeithertheCB1RantagonistSRl41716AortheCB2RantagonistAM630As

9、aresult,botllofSRl41716AandAM630wereabletosignificantlydecreasetheaveragelengthofneuritetreatedwithWINItsuggustedthatWINenhancedtheneuriteoutgrowthbyactivatingbothCB1RandCB2RTodelineatehowWINenhancedneudteoutgrowthweexam

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