星形膠質(zhì)細(xì)胞課件

NeuronNeuroglia(glia)

PNS:SchwanncellsSatellitecells

CNS:AstrocytesOligodendrocytesMicroglialcellsEpendymalcells

CompositionofNervousTissueNeuronCompositionofNervous11.Neuronshave

TWO

kindsof“processes”calledaxonsanddendrites，butGlialcellsonlyhave

ONE.2.Neurons

CANgenerateactionpotentials,Glialcells

CANNOT.however,Glia

dohavearestingpotential.

3.Neurons

HAVE

synapsesthatuseneurotransmitters.GlialcellsDONOT

havechemicalsynapses.

4.NeuronsDO

NOT

continuetodivide.Glialcells

DO

continuetodivide.Gliaaredifferentfromneurons:1.NeuronshaveTWOkindsof“2Importanceofglialcellsinnervoussystemfunction

isVASTLYunderestimated!Evidenceismounting:NurturingroleSignalingrole(Memory?)InflammationPainManymoretocome!Importanceofglialcellsinn3TherearetwomainkindsinPNSSchwanncells（雪旺細(xì)胞）Satellitecells（衛(wèi)星細(xì)胞）TherearetwomainkindsinPN4星形膠質(zhì)細(xì)胞課件5SchwanncellSchwanncell6TheSchwanncellmakingthemyelinsheath(M)aroundtheaxon(A)wascutatthelevelofthenucleus(N).E-endoneurium.Scale=300nm.(Rat,trigeminalnerve.)TheSchwanncellmakingthemy7Schwann細(xì)胞功能：形成周圍神經(jīng)系統(tǒng)的有髓神經(jīng)纖維的髓鞘合成和分泌多種營養(yǎng)因子、細(xì)胞外基質(zhì)成分和細(xì)胞黏附分子。周圍神經(jīng)損傷后的再生和修復(fù)Schwann細(xì)胞功能：形成周圍神經(jīng)系統(tǒng)的有髓神經(jīng)纖維的髓8TherearefourmainkindsinCNSAstrocytes(星形膠質(zhì)細(xì)胞）Oligodendrocytes(少突膠質(zhì)細(xì)胞）Ependymalcells

(室管膜細(xì)胞）Microglialcells

(小膠質(zhì)細(xì)胞）TherearefourmainkindsinC9星形膠質(zhì)細(xì)胞課件10Astrocytes

(AS,星形膠質(zhì)細(xì)胞）Astrocytes

(AS,星形膠質(zhì)細(xì)胞）11Mostnumerouscelltypeinbrain

Constitute~30-50%ofbrainvolumeSomeastrocytesdevelopprocesseswithexpandedend-feet

（終足）thatarelinkedtoendothelialcells,ortoependymalcells，etal.ExpressGFAP-anintermediatefilamentproteinProperties

ofASMostnumerouscelltypeinbra12突起的形態(tài)的不同：

纖維性星形細(xì)胞：白質(zhì)中，突起細(xì)長，分支少。

原漿性星形細(xì)胞：灰質(zhì)中，突起粗短，分支多?？乖瓨?biāo)記物的不同：

1型星形膠質(zhì)細(xì)胞：GFAP+/A2B5-2型星形膠質(zhì)細(xì)胞：GFAP+/A2B5+

分類突起的形態(tài)的不同：分類13FibrousastrocyteFibrousastrocyte14protoplasmicastrocyteprotoplasmicastrocyte15星形細(xì)胞的功能：為神經(jīng)細(xì)胞提供支持、隔離的作用合成和分泌多種細(xì)胞因子（黏連蛋白，生長因子和營養(yǎng)因子），參與神經(jīng)損傷后再生和修復(fù)；參與神經(jīng)元的遷移；參與血腦屏障的構(gòu)成參與內(nèi)環(huán)境的穩(wěn)定：具有多數(shù)神經(jīng)遞質(zhì)的受體，參與神經(jīng)元及其突觸功能的離子、遞質(zhì)和代謝的調(diào)節(jié),突觸的形成和可塑性星形細(xì)胞的功能：為神經(jīng)細(xì)胞提供支持、隔離的作用16功能：新的觀點(diǎn)：星形膠質(zhì)細(xì)胞負(fù)責(zé)大腦信息傳送

2007《science》“大腦中與記憶和學(xué)習(xí)有關(guān)的海馬區(qū)中的星形膠質(zhì)細(xì)胞能夠釋放一種谷氨酸化學(xué)信號，這種信號可以提高神經(jīng)元間的信號效率”

※星形膠質(zhì)細(xì)胞可以讓高度興奮的神經(jīng)元保持適當(dāng)“冷靜”，免受疾病侵襲；※星形膠質(zhì)細(xì)胞能使神經(jīng)元產(chǎn)生長時間增強(qiáng)反應(yīng)，使人學(xué)得進(jìn)、記得牢。

星形膠質(zhì)細(xì)胞與腦組織的神經(jīng)干細(xì)胞密切相關(guān)功能：新的觀點(diǎn)：星形膠質(zhì)細(xì)胞負(fù)責(zé)大腦信息傳送17Astrocytescanabsorblocalexcessofneurotransmitters,andreleasemetabolicandneuroactivemolecules.Astrocytescaninfluenceneuronalsurvivalandactivitythroughtheirabilitytoregulateconstituentsoftheextracellularenvironment。

AS參與維持中樞神經(jīng)系統(tǒng)內(nèi)環(huán)境穩(wěn)定Astrocytescanabsorblocalex18葡萄糖葡萄糖乳糖酵解normaldisease儲存糖原

1.參與葡萄糖代謝葡萄糖葡萄糖乳糖酵解normaldisease儲存糖原1.192.參與腦內(nèi)谷氨酸的代謝2.參與腦內(nèi)谷氨酸的代謝20星形膠質(zhì)細(xì)胞課件21Nature411,261-268(2001)

AChBPasanAChRdecoy星形膠質(zhì)細(xì)胞對突觸的調(diào)節(jié)作用Nature411,AChBPasa星形膠質(zhì)細(xì)胞對突22Astrocytes

AstrocytescontactvirtuallyeverycellcomponentinbrainOtherastrocytes(gapjunctions)EpendymalcellsNeurons(somas,processes,synapses)OligodendrogliaCapillaryendothelialcellsAstrocytes

Astrocytescontact23

肌萎縮側(cè)索硬化癥(ALS)

《nature》美國的兩個科研小組報告說:一種名為“神經(jīng)膠質(zhì)細(xì)胞”的變異神經(jīng)細(xì)胞所分泌的毒素，可導(dǎo)致肌萎縮側(cè)索硬化癥(ALS).

神經(jīng)系統(tǒng)疾病的關(guān)系肌萎縮側(cè)索硬化癥(ALS)《nature》美國的兩個24Oligodendrocytes

(少突膠質(zhì)細(xì)胞）束間少突膠質(zhì)細(xì)胞血管周少突膠質(zhì)細(xì)胞神經(jīng)元周少突膠質(zhì)細(xì)胞鍍銀染色發(fā)現(xiàn)突起很少；通過特異性的GC單抗做免疫細(xì)胞化學(xué)標(biāo)記，則可見少突細(xì)胞有很多突起，而且分支很多。根據(jù)在中樞神經(jīng)系統(tǒng)中的位置和分布：Oligodendrocytes

(少突膠質(zhì)細(xì)胞25功能形成中樞神經(jīng)系統(tǒng)的髓鞘抑制神經(jīng)元軸突的生長和再生（神經(jīng)突起生長抑制蛋白）攝取神經(jīng)遞質(zhì)，參與神經(jīng)系統(tǒng)內(nèi)的吞噬活動等功能形成中樞神經(jīng)系統(tǒng)的髓鞘26星形膠質(zhì)細(xì)胞課件27星形膠質(zhì)細(xì)胞課件28星形膠質(zhì)細(xì)胞課件29Ependymalcells

(室管膜細(xì)胞）室管膜細(xì)胞分布于腦室和脊髓中央管腔面，形似單層立方或柱狀上皮，細(xì)胞表面有纖毛和微絨毛Ependymalcells

(室管膜細(xì)胞）30EpendymalcellsEpendymalcells31Whoarethestemcellsoftheadultbrain?

astrocytesvs.ependymalcellsastrocytesEpendymalcellsJohanssonetal.,"Identificationofaneuralstemcellintheadultmammaliancentralnervoussystem,"Cell,96:25-34,1999.Doetschetal.,"Subventricularzoneastrocytesareneuralstem

cellsintheadultmammalianbrain,"Cell,97:703-16,1999.ThesubventricularzoneDrJonasFrisencontendsthatneuralstemcellsarecellsthatlineventriclesDrArturoAlvarez-Buyllacontendsneuralstemareastrocytesthatlieonelayerinfromventriclelining;Whoarethestemcellsofthe32Microglialcells

(小膠質(zhì)細(xì)胞）Microglialcells

(小膠質(zhì)細(xì)胞）33microgliamicroglia34Typicalfeaturesofneuronophagia,aclusterofmicrogliaaroundanecroticneuron,areshown(HE,highpower).Typicalfeaturesofneuronopha35Ontheleftsideofthediagram,cellslabeled"B1","B2",and"B3"representchronicallyactivatedmicroglialcells."B3"isamarkedlyelongatedcell,thenucleusinparticularhavingalong,rod-shapedform.Thisrodcellisclassicallyseeningeneralparesis(pareticneurosyphilis)butmayalsobeseeninotherchronicdiseasestatesofthebrain.Ontheleftsideofthediagra36功能吞噬功能靜止態(tài)——反應(yīng)態(tài)小膠質(zhì)細(xì)胞免疫功能神經(jīng)元變性——小膠質(zhì)細(xì)胞——表達(dá)MHC抗原——呈遞給T淋巴細(xì)胞——發(fā)揮免疫作用功能吞噬功能37膠質(zhì)細(xì)胞培養(yǎng)/content/pw6137808v577k16/fulltext.pdf

膠質(zhì)細(xì)胞培養(yǎng)http://www.springerlink.38材料儀器設(shè)備材料儀器設(shè)備39星形膠質(zhì)細(xì)胞課件40步驟PREPARATIONOFNEWBORNRATSFORCULTURING步驟PREPARATIONOFNEWBORNRATS41步驟(細(xì)胞懸液的制備）Dissection:1.Removebrainstem,cerebellumanddiencephalons.Immediatelyplaceincolddissectionbuffer2.Peeloffthemeningesandtransfercortextoatubecontainingcolddissectionbuffer.Immediatelyplaceonice3.Pourtissueintoadishandwash,usingasyringe,withmodifiedDMEM/F12culturemediumwith10%FBS,1%glutamine,andgentamicinantibiotic4MechanicallydigesttissuebymincingwithasterilerazorbladeTransfertissuebacktoatubecontaining5ml1Xtrypsinand50uLDNAse.Incubateat37oCfor25minutes.Removeonlytoswirltubeevery5minutes5.WashtissuewithGlialMediumtwice步驟(細(xì)胞懸液的制備）Dissection:426.Dissociatetissueintoacellsuspensionbygentlytitratingthrougha5mLpipettefollowedbyafire-polishedPasteurpipette.Transfersupernatanttoafreshtubeduringeachtitration.Doatotalofthreetitrationsusingbothpipettes7.Dilutesuspendedcellsin10mLofGlialMedium,andpassthesolutionthrougha40uMstrainer8.Centrifugecellsat1700rpmfor5minutes9.Removesupernatant,andresuspendpelletwith10mLGlialMediumSeed2x106cells/T75in15mlGlialmedium10.Incubatetheflasksat37oCin5%CO2for2-3dayswithoutdisruption.Changethemediumineachflaskevery2-3daysbyaspiratingandadding15mLfreshGlialMediumuntilconfluencyisachieved(afterapproximately6-7days)40uMstrainer步驟(細(xì)胞懸液的制備）6.Dissociatetissueintoace43步驟(純化）步驟(純化）44步驟(鑒定）CD11b/DAPI星形膠質(zhì)細(xì)胞小膠質(zhì)細(xì)胞步驟(鑒定）CD11b/DAPI星形膠質(zhì)細(xì)胞小膠質(zhì)細(xì)胞45少突膠質(zhì)細(xì)胞少突膠質(zhì)細(xì)胞46Thanks！Thanks！47NeuronNeuroglia(glia)

PNS:SchwanncellsSatellitecells

CNS:AstrocytesOligodendrocytesMicroglialcellsEpendymalcells

CompositionofNervousTissueNeuronCompositionofNervous481.Neuronshave

TWO

kindsof“processes”calledaxonsanddendrites，butGlialcellsonlyhave

ONE.2.Neurons

CANgenerateactionpotentials,Glialcells

CANNOT.however,Glia

dohavearestingpotential.

3.Neurons

HAVE

synapsesthatuseneurotransmitters.GlialcellsDONOT

havechemicalsynapses.

4.NeuronsDO

NOT

continuetodivide.Glialcells

DO

continuetodivide.Gliaaredifferentfromneurons:1.NeuronshaveTWOkindsof“49Importanceofglialcellsinnervoussystemfunction

isVASTLYunderestimated!Evidenceismounting:NurturingroleSignalingrole(Memory?)InflammationPainManymoretocome!Importanceofglialcellsinn50TherearetwomainkindsinPNSSchwanncells（雪旺細(xì)胞）Satellitecells（衛(wèi)星細(xì)胞）TherearetwomainkindsinPN51星形膠質(zhì)細(xì)胞課件52SchwanncellSchwanncell53TheSchwanncellmakingthemyelinsheath(M)aroundtheaxon(A)wascutatthelevelofthenucleus(N).E-endoneurium.Scale=300nm.(Rat,trigeminalnerve.)TheSchwanncellmakingthemy54Schwann細(xì)胞功能：形成周圍神經(jīng)系統(tǒng)的有髓神經(jīng)纖維的髓鞘合成和分泌多種營養(yǎng)因子、細(xì)胞外基質(zhì)成分和細(xì)胞黏附分子。周圍神經(jīng)損傷后的再生和修復(fù)Schwann細(xì)胞功能：形成周圍神經(jīng)系統(tǒng)的有髓神經(jīng)纖維的髓55TherearefourmainkindsinCNSAstrocytes(星形膠質(zhì)細(xì)胞）Oligodendrocytes(少突膠質(zhì)細(xì)胞）Ependymalcells

(室管膜細(xì)胞）Microglialcells

(小膠質(zhì)細(xì)胞）TherearefourmainkindsinC56星形膠質(zhì)細(xì)胞課件57Astrocytes

(AS,星形膠質(zhì)細(xì)胞）Astrocytes

(AS,星形膠質(zhì)細(xì)胞）58Mostnumerouscelltypeinbrain

Constitute~30-50%ofbrainvolumeSomeastrocytesdevelopprocesseswithexpandedend-feet

（終足）thatarelinkedtoendothelialcells,ortoependymalcells，etal.ExpressGFAP-anintermediatefilamentproteinProperties

ofASMostnumerouscelltypeinbra59突起的形態(tài)的不同：

纖維性星形細(xì)胞：白質(zhì)中，突起細(xì)長，分支少。

原漿性星形細(xì)胞：灰質(zhì)中，突起粗短，分支多?？乖瓨?biāo)記物的不同：

1型星形膠質(zhì)細(xì)胞：GFAP+/A2B5-2型星形膠質(zhì)細(xì)胞：GFAP+/A2B5+

分類突起的形態(tài)的不同：分類60FibrousastrocyteFibrousastrocyte61protoplasmicastrocyteprotoplasmicastrocyte62星形細(xì)胞的功能：為神經(jīng)細(xì)胞提供支持、隔離的作用合成和分泌多種細(xì)胞因子（黏連蛋白，生長因子和營養(yǎng)因子），參與神經(jīng)損傷后再生和修復(fù)；參與神經(jīng)元的遷移；參與血腦屏障的構(gòu)成參與內(nèi)環(huán)境的穩(wěn)定：具有多數(shù)神經(jīng)遞質(zhì)的受體，參與神經(jīng)元及其突觸功能的離子、遞質(zhì)和代謝的調(diào)節(jié),突觸的形成和可塑性星形細(xì)胞的功能：為神經(jīng)細(xì)胞提供支持、隔離的作用63功能：新的觀點(diǎn)：星形膠質(zhì)細(xì)胞負(fù)責(zé)大腦信息傳送

2007《science》“大腦中與記憶和學(xué)習(xí)有關(guān)的海馬區(qū)中的星形膠質(zhì)細(xì)胞能夠釋放一種谷氨酸化學(xué)信號，這種信號可以提高神經(jīng)元間的信號效率”

※星形膠質(zhì)細(xì)胞可以讓高度興奮的神經(jīng)元保持適當(dāng)“冷靜”，免受疾病侵襲；※星形膠質(zhì)細(xì)胞能使神經(jīng)元產(chǎn)生長時間增強(qiáng)反應(yīng)，使人學(xué)得進(jìn)、記得牢。

星形膠質(zhì)細(xì)胞與腦組織的神經(jīng)干細(xì)胞密切相關(guān)功能：新的觀點(diǎn)：星形膠質(zhì)細(xì)胞負(fù)責(zé)大腦信息傳送64Astrocytescanabsorblocalexcessofneurotransmitters,andreleasemetabolicandneuroactivemolecules.Astrocytescaninfluenceneuronalsurvivalandactivitythroughtheirabilitytoregulateconstituentsoftheextracellularenvironment。

AS參與維持中樞神經(jīng)系統(tǒng)內(nèi)環(huán)境穩(wěn)定Astrocytescanabsorblocalex65葡萄糖葡萄糖乳糖酵解normaldisease儲存糖原

1.參與葡萄糖代謝葡萄糖葡萄糖乳糖酵解normaldisease儲存糖原1.662.參與腦內(nèi)谷氨酸的代謝2.參與腦內(nèi)谷氨酸的代謝67星形膠質(zhì)細(xì)胞課件68Nature411,261-268(2001)

AChBPasanAChRdecoy星形膠質(zhì)細(xì)胞對突觸的調(diào)節(jié)作用Nature411,AChBPasa星形膠質(zhì)細(xì)胞對突69Astrocytes

AstrocytescontactvirtuallyeverycellcomponentinbrainOtherastrocytes(gapjunctions)EpendymalcellsNeurons(somas,processes,synapses)OligodendrogliaCapillaryendothelialcellsAstrocytes

Astrocytescontact70

肌萎縮側(cè)索硬化癥(ALS)

《nature》美國的兩個科研小組報告說:一種名為“神經(jīng)膠質(zhì)細(xì)胞”的變異神經(jīng)細(xì)胞所分泌的毒素，可導(dǎo)致肌萎縮側(cè)索硬化癥(ALS).

神經(jīng)系統(tǒng)疾病的關(guān)系肌萎縮側(cè)索硬化癥(ALS)《nature》美國的兩個71Oligodendrocytes

(少突膠質(zhì)細(xì)胞）束間少突膠質(zhì)細(xì)胞血管周少突膠質(zhì)細(xì)胞神經(jīng)元周少突膠質(zhì)細(xì)胞鍍銀染色發(fā)現(xiàn)突起很少；通過特異性的GC單抗做免疫細(xì)胞化學(xué)標(biāo)記，則可見少突細(xì)胞有很多突起，而且分支很多。根據(jù)在中樞神經(jīng)系統(tǒng)中的位置和分布：Oligodendrocytes

(少突膠質(zhì)細(xì)胞72功能形成中樞神經(jīng)系統(tǒng)的髓鞘抑制神經(jīng)元軸突的生長和再生（神經(jīng)突起生長抑制蛋白）攝取神經(jīng)遞質(zhì)，參與神經(jīng)系統(tǒng)內(nèi)的吞噬活動等功能形成中樞神經(jīng)系統(tǒng)的髓鞘73星形膠質(zhì)細(xì)胞課件74星形膠質(zhì)細(xì)胞課件75星形膠質(zhì)細(xì)胞課件76Ependymalcells

(室管膜細(xì)胞）室管膜細(xì)胞分布于腦室和脊髓中央管腔面，形似單層立方或柱狀上皮，細(xì)胞表面有纖毛和微絨毛Ependymalcells

(室管膜細(xì)胞）77EpendymalcellsEpendymalcells78Whoarethestemcellsoftheadultbrain?

astrocytesvs.ependymalcellsastrocytesEpendymalcellsJohanssonetal.,"Identificationofaneuralstemcellintheadultmammaliancentralnervoussystem,"Cell,96:25-34,1999.Doetschetal.,"Subventricularzoneastrocytesareneuralstem

cellsintheadultmammalianbrain,"Cell,97:703-16,1999.ThesubventricularzoneDrJonasFrisencontendsthatneuralstemcellsarecellsthatlineventriclesDrArturoAlvarez-Buyllacontendsneuralstemareastrocytesthatlieonelayerinfromventriclelining;Whoarethestemcellsofthe79Microglialcells

(小膠質(zhì)細(xì)胞）Microglialcells

(小膠質(zhì)細(xì)胞）80microgliamicroglia81Typicalfeaturesofneuronophagia,aclusterofmicrogliaaroundanecroticneuron,areshown(HE,highpower).Typicalfeaturesofneuronopha82Ontheleftsideofthediagram,cellslabeled"B1","B2",and"B3"representchronicallyactivatedmicroglialcells."B3"isamarkedlyelongatedcell,thenucleusinparticularhavingalong,rod-shapedform.Thisrodcellisclassicallyseeningeneralparesis(pareticneurosyphilis)butmayalsobeseeninotherchronicdiseasestatesofthebrain.Ontheleftsideofthediagra83功能吞噬功能靜止態(tài)——反應(yīng)態(tài)小膠質(zhì)細(xì)胞免疫功能神經(jīng)元變性——小膠質(zhì)細(xì)胞——表達(dá)MHC抗原——呈遞給T淋巴細(xì)胞——發(fā)揮免疫作用功能吞噬功能84膠質(zhì)細(xì)胞培養(yǎng)/content/pw6137808v577k16/fulltext.pdf

膠質(zhì)細(xì)胞培養(yǎng)http://www.springerlink.85材料儀器設(shè)備材料儀器設(shè)備86星形膠質(zhì)細(xì)胞課件87步驟PREPARATIONOFNEWBORNRATSFORCULTURING步驟PREPARATIONOFNEWBORNRATS88步驟(細(xì)胞懸液的制備）Dissection:1.Removebrainstem,cerebellumanddiencephalons.Immediatelyplaceincolddissectionbuffer2.Peeloffthemeningesandtransf