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（雪旺細胞）Satellitecells（衛星細胞）TherearetwomainkindsinPN4星形膠質細胞課件5SchwanncellSchwanncell6TheSchwanncellmakingthemyelinsheath(M)aroundtheaxon(A)wascutatthelevelofthenucleus(N).E-endoneurium.Scale=300nm.(Rat,trigeminalnerve.)TheSchwanncellmakingthemy7Schwann細胞功能：形成周圍神經系統的有髓神經纖維的髓鞘合成和分泌多種營養因子、細胞外基質成分和細胞黏附分子。周圍神經損傷后的再生和修復Schwann細胞功能：形成周圍神經系統的有髓神經纖維的髓8TherearefourmainkindsinCNSAstrocytes(星形膠質細胞）Oligodendrocytes(少突膠質細胞）Ependymalcells

(室管膜細胞）Microglialcells

(小膠質細胞）TherearefourmainkindsinC9星形膠質細胞課件10Astrocytes

(AS,星形膠質細胞）Astrocytes

(AS,星形膠質細胞）11Mostnumerouscelltypeinbrain

Constitute~30-50%ofbrainvolumeSomeastrocytesdevelopprocesseswithexpandedend-feet

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

ofASMostnumerouscelltypeinbra12突起的形態的不同：

纖維性星形細胞：白質中，突起細長，分支少。

原漿性星形細胞：灰質中，突起粗短，分支多。抗原標記物的不同：

1型星形膠質細胞：GFAP+/A2B5-2型星形膠質細胞：GFAP+/A2B5+

分類突起的形態的不同：分類13FibrousastrocyteFibrousastrocyte14protoplasmicastrocyteprotoplasmicastrocyte15星形細胞的功能：為神經細胞提供支持、隔離的作用合成和分泌多種細胞因子（黏連蛋白，生長因子和營養因子），參與神經損傷后再生和修復；參與神經元的遷移；參與血腦屏障的構成參與內環境的穩定：具有多數神經遞質的受體，參與神經元及其突觸功能的離子、遞質和代謝的調節,突觸的形成和可塑性星形細胞的功能：為神經細胞提供支持、隔離的作用16功能：新的觀點：星形膠質細胞負責大腦信息傳送

2007《science》“大腦中與記憶和學習有關的海馬區中的星形膠質細胞能夠釋放一種谷氨酸化學信號，這種信號可以提高神經元間的信號效率”

※星形膠質細胞可以讓高度興奮的神經元保持適當“冷靜”，免受疾病侵襲；※星形膠質細胞能使神經元產生長時間增強反應，使人學得進、記得牢。

星形膠質細胞與腦組織的神經干細胞密切相關功能：新的觀點：星形膠質細胞負責大腦信息傳送17Astrocytescanabsorblocalexcessofneurotransmitters,andreleasemetabolicandneuroactivemolecules.Astrocytescaninfluenceneuronalsurvivalandactivitythroughtheirabilitytoregulateconstituentsoftheextracellularenvironment。

AS參與維持中樞神經系統內環境穩定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1.192.參與腦內谷氨酸的代謝2.參與腦內谷氨酸的代謝20星形膠質細胞課件21Nature411,261-268(2001)

AChBPasanAChRdecoy星形膠質細胞對突觸的調節作用Nature411,AChBPasa星形膠質細胞對突22Astrocytes

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

Astrocytescontact23

肌萎縮側索硬化癥(ALS)

《nature》美國的兩個科研小組報告說:一種名為“神經膠質細胞”的變異神經細胞所分泌的毒素，可導致肌萎縮側索硬化癥(ALS).

神經系統疾病的關系肌萎縮側索硬化癥(ALS)《nature》美國的兩個24Oligodendrocytes

(少突膠質細胞）束間少突膠質細胞血管周少突膠質細胞神經元周少突膠質細胞鍍銀染色發現突起很少；通過特異性的GC單抗做免疫細胞化學標記，則可見少突細胞有很多突起，而且分支很多。根據在中樞神經系統中的位置和分布：Oligodendrocytes

(少突膠質細胞25功能形成中樞神經系統的髓鞘抑制神經元軸突的生長和再生（神經突起生長抑制蛋白）攝取神經遞質，參與神經系統內的吞噬活動等功能形成中樞神經系統的髓鞘26星形膠質細胞課件27星形膠質細胞課件28星形膠質細胞課件29Ependymalcells

(室管膜細胞）室管膜細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，細胞表面有纖毛和微絨毛Ependymalcells

(室管膜細胞）30EpendymalcellsEpendymalcells31Whoarethestemcellsoftheadultbrain?

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

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

(小膠質細胞）Microglialcells

(小膠質細胞）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——反應態小膠質細胞免疫功能神經元變性——小膠質細胞——表達MHC抗原——呈遞給T淋巴細胞——發揮免疫作用功能吞噬功能37膠質細胞培養/content/pw6137808v577k16/fulltext.pdf

膠質細胞培養http://www.springerlink.38材料儀器設備材料儀器設備39星形膠質細胞課件40步驟PREPARATIONOFNEWBORNRATSFORCULTURING步驟PREPARATIONOFNEWBORNRATS41步驟(細胞懸液的制備）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星形膠質細胞小膠質細胞步驟(鑒定）CD11b/DAPI星形膠質細胞小膠質細胞45少突膠質細胞少突膠質細胞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（雪旺細胞）Satellitecells（衛星細胞）TherearetwomainkindsinPN51星形膠質細胞課件52SchwanncellSchwanncell53TheSchwanncellmakingthemyelinsheath(M)aroundtheaxon(A)wascutatthelevelofthenucleus(N).E-endoneurium.Scale=300nm.(Rat,trigeminalnerve.)TheSchwanncellmakingthemy54Schwann細胞功能：形成周圍神經系統的有髓神經纖維的髓鞘合成和分泌多種營養因子、細胞外基質成分和細胞黏附分子。周圍神經損傷后的再生和修復Schwann細胞功能：形成周圍神經系統的有髓神經纖維的髓55TherearefourmainkindsinCNSAstrocytes(星形膠質細胞）Oligodendrocytes(少突膠質細胞）Ependymalcells

(室管膜細胞）Microglialcells

(小膠質細胞）TherearefourmainkindsinC56星形膠質細胞課件57Astrocytes

(AS,星形膠質細胞）Astrocytes

(AS,星形膠質細胞）58Mostnumerouscelltypeinbrain

Constitute~30-50%ofbrainvolumeSomeastrocytesdevelopprocesseswithexpandedend-feet

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

ofASMostnumerouscelltypeinbra59突起的形態的不同：

纖維性星形細胞：白質中，突起細長，分支少。

原漿性星形細胞：灰質中，突起粗短，分支多。抗原標記物的不同：

1型星形膠質細胞：GFAP+/A2B5-2型星形膠質細胞：GFAP+/A2B5+

分類突起的形態的不同：分類60FibrousastrocyteFibrousastrocyte61protoplasmicastrocyteprotoplasmicastrocyte62星形細胞的功能：為神經細胞提供支持、隔離的作用合成和分泌多種細胞因子（黏連蛋白，生長因子和營養因子），參與神經損傷后再生和修復；參與神經元的遷移；參與血腦屏障的構成參與內環境的穩定：具有多數神經遞質的受體，參與神經元及其突觸功能的離子、遞質和代謝的調節,突觸的形成和可塑性星形細胞的功能：為神經細胞提供支持、隔離的作用63功能：新的觀點：星形膠質細胞負責大腦信息傳送

2007《science》“大腦中與記憶和學習有關的海馬區中的星形膠質細胞能夠釋放一種谷氨酸化學信號，這種信號可以提高神經元間的信號效率”

※星形膠質細胞可以讓高度興奮的神經元保持適當“冷靜”，免受疾病侵襲；※星形膠質細胞能使神經元產生長時間增強反應，使人學得進、記得牢。

星形膠質細胞與腦組織的神經干細胞密切相關功能：新的觀點：星形膠質細胞負責大腦信息傳送64Astrocytescanabsorblocalexcessofneurotransmitters,andreleasemetabolicandneuroactivemolecules.Astrocytescaninfluenceneuronalsurvivalandactivitythroughtheirabilitytoregulateconstituentsoftheextracellularenvironment。

AS參與維持中樞神經系統內環境穩定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1.662.參與腦內谷氨酸的代謝2.參與腦內谷氨酸的代謝67星形膠質細胞課件68Nature411,261-268(2001)

AChBPasanAChRdecoy星形膠質細胞對突觸的調節作用Nature411,AChBPasa星形膠質細胞對突69Astrocytes

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

Astrocytescontact70

肌萎縮側索硬化癥(ALS)

《nature》美國的兩個科研小組報告說:一種名為“神經膠質細胞”的變異神經細胞所分泌的毒素，可導致肌萎縮側索硬化癥(ALS).

神經系統疾病的關系肌萎縮側索硬化癥(ALS)《nature》美國的兩個71Oligodendrocytes

(少突膠質細胞）束間少突膠質細胞血管周少突膠質細胞神經元周少突膠質細胞鍍銀染色發現突起很少；通過特異性的GC單抗做免疫細胞化學標記，則可見少突細胞有很多突起，而且分支很多。根據在中樞神經系統中的位置和分布：Oligodendrocytes

(少突膠質細胞72功能形成中樞神經系統的髓鞘抑制神經元軸突的生長和再生（神經突起生長抑制蛋白）攝取神經遞質，參與神經系統內的吞噬活動等功能形成中樞神經系統的髓鞘73星形膠質細胞課件74星形膠質細胞課件75星形膠質細胞課件76Ependymalcells

(室管膜細胞）室管膜細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，細胞表面有纖毛和微絨毛Ependymalcells

(室管膜細胞）77EpendymalcellsEpendymalcells78Whoarethestemcellsoftheadultbrain?

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

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

(小膠質細胞）Microglialcells

(小膠質細胞）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——反應態小膠質細胞免疫功能神經元變性——小膠質細胞——表達MHC抗原——呈遞給T淋巴細胞——發揮免疫作用功能吞噬功能84膠質細胞培養/content/pw6137808v577k16/fulltext.pdf

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