为大脑半球结构中主要的灰质部分之一课件

IntrastriataltargetsofprojectionfibersfromthecentrallateralnucleusoftheratthalamusNoritakalchinohe,Hiroyasulwatsuki,KazuhikoShoumuraNeuroscienceLetters302(2001)105-108Intrastriataltargetsofproje1IntroductionThebasalgangliaandthecerebellumarethemajorsubcorticalstructuresthatareimportantforregulationandcontrolofvoluntarymovementProjectionfromthedeepcerebellarnucleitothemotor-relatedStriatum,whichreceivesinputsfromthemotorcorticalareasHavebeenreportedtoberelayedatseveralthalamicnuclei;theventrallateralnucleusparscaudails,ventralposteriorlateralnucleusparsoralis,areaXandcentrallateralnucleus(CL)inthemonkey,andtheventrallateralnucleusandCLintheratIntroductionThebasalganglia2Basalganglia為大腦半球結構中主要的灰質部分之一，也就是neuron之所在分為：Caudatenucleus（尾狀核）&Putamen(被殼）Basalganglia為大腦半球結構中主要的灰質部分之一3StriatumCaudatenucleus&putamen合稱striatum（紋狀體）因經過的axon把他們弄出了條紋的外觀臨床上，basalganglia是指striatum,視丘下部神經核,黑質與運動功能異常有關，特徵，不自主，無目的的運動StriatumCaudatenucleus&puta4IntroductionPreciseunderstandingofthefunctionofthesecerebello-thalamo-striatalprojectionsrequiresdetailedknowledgeofintrastriataltargetsofprojectionfibersfromthecerebello-recipientthalamicnucleiIntroductionPreciseunderstand5InthepreviousstudyWedemostratedthatthalamostriatalneuronsintheCLreceivedirectstrongprojectionsfromthelateralcerebellarnucleusintheratInthepreviousstudyWedemost6Specialattentiontoa.whetherornotprojectionfibersfromtheCLmakesynapsesondendriticspines,whichareagoodmakerforstriatalprojectionneuronsb.whetherornotprojectionfibersfromtheCLareinsynapticcontactwithparvalbumin(PV)immunopostivestriatalinterneuron,whichrepresentthemajorclassofGABAergicinterneuronsinthestriatumSpecialattentiontoa.whether7Methoda.TenmaleWistarratswereanesthetizedwithsodiumPentobarbital(70mg/kg,I.P)b.10%solutionofbiotindextranamine(BDA)in0.01MPB(pH7.2)wasiontophoreticallyinjectedintotheCLc.Theglassmicropipette(tipdiameter:15μm)usingapositivecurrent(5-7μA,7son/7soff)for20~30minsMethoda.TenmaleWistarratsw8SurvivalTheperiodof7days,theratswerere-anesthetizedandperfusedtranscardiallywith100mlofsalinefollowedby500mlofparaformaldehydeand0.1%glutaraldehydein0.1MPB(pH7.4)Post-fixedinthesamefixativeandstoredovernightat4℃inPB,thefollowingday,thebrainwasseriallycut50μmthickonamicroslicerSurvivalTheperiodof7days,t9Staining(fordextran)1.wash2.ABCkit(1:100~1:200dilute)for2hatroomtemperature3.wash4.0.1%DABin0.05MTrisand0.003%H2O2BDA-labeledaxonswerevisualizedasbrownStaining(fordextran)1.wash10Staining(forPV)PVimmunohistochemistryusingVIPasachromogen1.wash2.mouseanti-PVmonoclonalantibodydiluted1:1000for24hat4℃3.1:200biotinylatedhorseanti-mouseIgGantibody2h4.1:100ABCkitfor2h5.VIPkitPVimmunopositivestructureswerevisualizedaspurpleStaining(forPV)PVimmunohisto11ResultInjectionsiteResultInjectionsite12ToevaluteTherelativefrequencyoftheCLboutonstoallexcitatoryinputsonPVimmunopositiveinterneurons,werecordedbothBDA-labeledandunlabeledboutonswhichmadeasymmerticalsynapticspecializationswithPVimmunopositivestructuresToevaluteTherelativefrequen13ResultBDA-labeledfibersweredistributedmainlyinthelaterodorsalpartofthestriatumandintermingledwithPVimmunopositiveneuronsAllBDA-labeledfibersweresmallindiameter(0.1~0.3μm)andexhibitedanextendedtypeofarborizationwithlong,straight,relativelyunbranchedtrajectoriesthroughneuropilResultBDA-labeledfiberswere14为大脑半球结构中主要的灰质部分之一课件15ResultOf408BDA-labeledterminals,371(90.9%)wereinsynapticcontractwithPVimmunonegativedendriticspines,35(8.6%)madesynapsesonPVimmunonegativedendriticshafts,andonlytwo(0.5%)formedsynapsesonPVimmunopositivedendriticshaftsResultOf408BDA-labeledtermi16DiscussionInthepresentstudy,wedemostratedthatalargeproportion(91%)ofthalamostriatalfibersfromtheCLterminatedonPVimmunonegativedendriticspines,whichwereassumedtobethoseofstriatalprojectionneuronsDiscussionInthepresentstudy17ThusThetargetpreferenceofthalamostriatalfibersfromtheCLseemsnottohaveasignificantbiastointerneuronsoverprojectionneuronsinthestriatumThusThetargetpreferenceoft18InsummaryThecerebello-CL-striatalprojectionhavepredominantaccess(probablyexcitatory)tostriatalprojectionneurons,andalsominoraccesstoPVimmunopositivestriatalinterneurons,whichareastrongGABAnergicinhibitorysourcetostriatalprojectionneuronsInsummaryThecerebello-CL-str19GABAGABA神經元的作用是在提供負回饋迴路(negativefeedbackloop)目的，增加運動控制系統的穩定性GABAGABA神經元的作用是在提供負回饋迴路20IntrastriataltargetsofprojectionfibersfromthecentrallateralnucleusoftheratthalamusNoritakalchinohe,Hiroyasulwatsuki,KazuhikoShoumuraNeuroscienceLetters302(2001)105-108Intrastriataltargetsofproje21IntroductionThebasalgangliaandthecerebellumarethemajorsubcorticalstructuresthatareimportantforregulationandcontrolofvoluntarymovementProjectionfromthedeepcerebellarnucleitothemotor-relatedStriatum,whichreceivesinputsfromthemotorcorticalareasHavebeenreportedtoberelayedatseveralthalamicnuclei;theventrallateralnucleusparscaudails,ventralposteriorlateralnucleusparsoralis,areaXandcentrallateralnucleus(CL)inthemonkey,andtheventrallateralnucleusandCLintheratIntroductionThebasalganglia22Basalganglia為大腦半球結構中主要的灰質部分之一，也就是neuron之所在分為：Caudatenucleus（尾狀核）&Putamen(被殼）Basalganglia為大腦半球結構中主要的灰質部分之一23StriatumCaudatenucleus&putamen合稱striatum（紋狀體）因經過的axon把他們弄出了條紋的外觀臨床上，basalganglia是指striatum,視丘下部神經核,黑質與運動功能異常有關，特徵，不自主，無目的的運動StriatumCaudatenucleus&puta24IntroductionPreciseunderstandingofthefunctionofthesecerebello-thalamo-striatalprojectionsrequiresdetailedknowledgeofintrastriataltargetsofprojectionfibersfromthecerebello-recipientthalamicnucleiIntroductionPreciseunderstand25InthepreviousstudyWedemostratedthatthalamostriatalneuronsintheCLreceivedirectstrongprojectionsfromthelateralcerebellarnucleusintheratInthepreviousstudyWedemost26Specialattentiontoa.whetherornotprojectionfibersfromtheCLmakesynapsesondendriticspines,whichareagoodmakerforstriatalprojectionneuronsb.whetherornotprojectionfibersfromtheCLareinsynapticcontactwithparvalbumin(PV)immunopostivestriatalinterneuron,whichrepresentthemajorclassofGABAergicinterneuronsinthestriatumSpecialattentiontoa.whether27Methoda.TenmaleWistarratswereanesthetizedwithsodiumPentobarbital(70mg/kg,I.P)b.10%solutionofbiotindextranamine(BDA)in0.01MPB(pH7.2)wasiontophoreticallyinjectedintotheCLc.Theglassmicropipette(tipdiameter:15μm)usingapositivecurrent(5-7μA,7son/7soff)for20~30minsMethoda.TenmaleWistarratsw28SurvivalTheperiodof7days,theratswerere-anesthetizedandperfusedtranscardiallywith100mlofsalinefollowedby500mlofparaformaldehydeand0.1%glutaraldehydein0.1MPB(pH7.4)Post-fixedinthesamefixativeandstoredovernightat4℃inPB,thefollowingday,thebrainwasseriallycut50μmthickonamicroslicerSurvivalTheperiodof7days,t29Staining(fordextran)1.wash2.ABCkit(1:100~1:200dilute)for2hatroomtemperature3.wash4.0.1%DABin0.05MTrisand0.003%H2O2BDA-labeledaxonswerevisualizedasbrownStaining(fordextran)1.wash30Staining(forPV)PVimmunohistochemistryusingVIPasachromogen1.wash2.mouseanti-PVmonoclonalantibodydiluted1:1000for24hat4℃3.1:200biotinylatedhorseanti-mouseIgGantibody2h4.1:100ABCkitfor2h5.VIPkitPVimmunopositivestructureswerevisualizedaspurpleStaining(forPV)PVimmunohisto31ResultInjectionsiteResultInjectionsite32ToevaluteTherelativefrequencyoftheCLboutonstoallexcitatoryinputsonPVimmunopositiveinterneurons,werecordedbothBDA-labeledandunlabeledboutonswhichmadeasymmerticalsynapticspecializationswithPVimmunopositivestructuresToevaluteTherelativefrequen33ResultBDA-labeledfibersweredistributedmainlyinthelaterodorsalpartofthestriatumandintermingledwithPVimmunopositiveneuronsAllBDA-labeledfibersweresmallindiameter(0.1~0.3μm)andexhibitedanextendedtypeofarborizationwithlong,straight,relativelyunbranchedtrajectoriesthroughneuropilResultBDA-labeledfiberswere34为大脑半球结构中主要的灰质部分之一课件35ResultOf408BDA-labeledterminals,371(90.9%)wereinsynapticcontractwithPVimmunonegativedendriticspines,35(8.6%)madesynapses