乙酰胆碱和其他神经递质专家讲座

乙酰胆碱和其它神经递质ZhuCui-Qing.101乙酰胆碱和其他神经递质第1页Chapter6:Acetylcholine

乙酰胆碱2乙酰胆碱和其他神经递质第2页GeneralknowledgeHistoryofAChasaneurotransmitter:1867:Chemicallysynthesized.乙酰胆碱3Dale(1914):theeffectofAChontheheartissimilarwiththatofvagalstimulation

Loewi(1921)、Dale（1924):infusionexperimentoffrogheart.1936:NobelPrize.乙酰胆碱和其他神经递质第3页Conceptofcholinergicneurons:Neuronsthatuseacetylcholineastheneurotransmitter.GeneralknowledgeChemicalpropertyofacetylcholine：a.Easilyhydrolyzed:stableatpH3.8-4.5；completehydrolysisatpH10and100°C.b.Easilycatalyticallyhydrolyzedbycholinesterase.生物学作用即时性生物样品保留和检测4乙酰胆碱和其他神经递质第4页Indirectmethods：a.Measureofcholinesteraseactivity：Acetylthiocholine(乙酰硫代胆碱法,AChE法）.b.ChAT:1970’s,ChATmultiplecolonialantibody;Eckenstein(1981)ChATmonoclonalantibody.Houser(1983):ConsistenceofAchEmethodandChATmethod.DirectmethodsRIA;ELISAChromatogram;HPLCetal.Imunohistology+fluorescenttracing.MeasureofacetylcholineGeneralknowledge5乙酰胆碱和其他神经递质第5页6SectionI:DistributionandprojectionofcholinergicneuronsintheCNS分布与投射Classificationofcentralcholinergicneurons：ProjectionneuronLocalcircuitneuronACh神经元分布在哪里？乙酰胆碱和其他神经递质第6页(I)基底前脑胆碱能系统(basalforebraincholinergicsystem)Mesulam和Perry分群：ch1-ch6。Ch1、Ch2、Ch3分别相当于隔内侧核（medialseptalnucleus）、斜角带垂直部（verticalnucleusofthediagonalband）、斜角带水平部（horizontallimbofthediagonalband）；Ch4则包含了视前大细胞区、苍白球底板无名质及其向前延伸苍白球腹侧区（纹状体下灰质）、Meynert基底核（nucleusbasalisofMeynert）、豆状攀核等界限并不很清楚区域。I.胆碱能投射神经元(Projectionneuron)7乙酰胆碱和其他神经递质第7页基底前脑胆碱能系统(basalforebraincholinergicsystem)主要投射通路：它们投射纤维主要形成以下通路：①隔内侧核、斜角带-海马通路；②斜角带-杏仁复合体通路；③隔区-僵内侧核（medialhabenula）、中脑脚间核通路；④斜角带、Meynert基底核-大脑皮层通路。其中由Meynert基底核向皮质额叶、颞叶、顶叶和视皮层胆碱能投射，与学习和记忆功效亲密相关。①②③④8④乙酰胆碱和其他神经递质第8页(II)脑干胆碱能系统(brainstemcholinergicsystem)

由脑桥被盖胆碱能系统（pontinetegmentalcholinergicsystem）和延髓胆碱能神经元组成。纤维：背侧被盖束和腹侧被盖束，向头端投射至丘脑、丘脑下部、苍白球、尾壳核等，并与其它上行纤维一起组成网状上行激活系统，引发觉醒和警觉。脑桥被盖胆碱能系统主要包含脚间脑桥被盖区（pedunculopontinetegmentalregion,ppt）和背外侧或外侧被盖核（dorsolateralorlaterodorsaltegmentalnucleus,ldt）中胆碱能神经元，相当于Mesulam和Perry分群中Ch5、Ch6。I.胆碱能投射神经元(Projectionneuron)9乙酰胆碱和其他神经递质第9页10乙酰胆碱和其他神经递质第10页中脑和脑桥动眼神经（III）、滑车神经（IV）、外展神经（VI）胆碱能运动神经元胞体。延髓胆碱能神经元主要分布：舌下神经核（XII）、迷走神经背核与疑核（X）、面神经核（VII）、三叉神经脊束核（V）11乙酰胆碱和其他神经递质第11页I.胆碱能投射神经元(Projectionneuron)(III)Cholinergicneuronsinthespinalcord脊髓前脚运动神经元侧角交感和副交感节前神经元12乙酰胆碱和其他神经递质第12页局部环路神经元这类神经元在核团内局部组成环路，不向核外发出投射，属中间神经元。纹状体（尾-壳核复合体，Caudate-putamencomplex）、伏隔核(nucleusaccumben)、嗅结节（olfactorytubercle）、海马和大脑皮质内Ⅱ-Ⅴ层。纹状体内胆碱能神经元主要为大、中型无棘突多极神经元，参加黑质-纹状体多巴胺系统对运动调整，其异常与帕金森氏病（Parkinson’sDisease,PD）病理过程亲密相关。孤束核、前庭外侧核、外侧网状核、巨细胞网状核、中缝大核等。脊髓背角胆碱能神经元投射到脊髓RexedⅡ-III层、小脑内第Ⅱ层颗粒细胞投射到Ⅴ层细胞也形成局部环路。II.Localcircuitneuron13乙酰胆碱和其他神经递质第13页基底前脑乙酰胆碱能系统(basalforebraincholinergicsystem)主要投射通路有哪些？皮层和海马是否有乙酰胆碱能神经元？Question

114乙酰胆碱和其他神经递质第14页-S-(I)BiosynthesisCholineAcetylcoenzymeACholineacetyltransfease胆碱乙酰转移酶Cholineacetylase胆碱乙酰化酶Substrates:胆碱乙酰辅酶ASectionII:biosynthesis,storageandrelease15乙酰胆碱和其他神经递质第15页ChAT,globulin,MW68000.Synthesizedincellbody,takeseffectinterminals.Essentialchemicalgroups:imidazole,-SH.Enzymedynamics(Km)：choline7.5×10-4mol/L;AcetylCoA1.0×10-5mol/L.Siteofsynthesis合成部位：CholinergicterminalsEnzyme:(I)Biosynthesis16乙酰胆碱和其他神经递质第16页ChAT咪唑基乙酰化转乙酰作用活性中心：咪唑基巯基AcetylcoenzymeACholine(I)BiosynthesisCholineacetyltransfease胆碱乙酰转移酶ChATCholineacetylase胆碱乙酰化酶17乙酰胆碱和其他神经递质第17页1.AcetylCoA.Source:(1)glucose-pyruvicacid-AcetylCoA.

（2）Fattyacidb-oxidation.

（3）synthesizedfromcitricacid.(4)acetate(non-neuraltissues)Substrate:(I)Biosynthesis18乙酰胆碱和其他神经递质第18页2.CholineSource:

a.uptakeandhydrolyzelecithinfromcirculation.b.hydrolysisoflecithinofneuronaltissues.c.hydrolysisofAchandreuptakeofcholine.1/3-1/2.卵磷脂(I)BiosynthesisSubstrate:19乙酰胆碱和其他神经递质第19页

Lowaffinitycarrier:distributedinallneuronsandgliacells.Km:40-100micromol/L.facilitateddiffusionTransportationofcholine:Highaffinitycarrierspecificallydistributedincholinergicterminals.Km:0.4-4micromol/LNa+,ATP-dependentactivetransportation.Satureduptake.Themosteffectivelimitfactorofacetylcholinesynthesis(!).Cholineisthelimitsubstrate.(I)Biosynthesis20乙酰胆碱和其他神经递质第20页Hemicholium-3(HC-3)密胆碱Mechanism：competehighaffinitycarrierwithcholine.Antagonizedbycholine.

神经末梢characteristics：unabletopassthoughblood-brainbarrier;centraladministrationneeded.Strong,slow,lastingeffect.Triethylcholine三乙基胆碱(CH3CH2)3N+-(CH2)2-OHMechanism：synthesisoffakeneurotransmitter，inadditiontocompetingwithcholineforuptake4-(1-naphthylvinyl)pyridine]

4-(1-萘乙烯基)吡啶Mechanism：：specificChATinhibitorinvitro.AgentsaffectingthebiosynthesisofACh(I)Biosynthesis21乙酰胆碱和其他神经递质第21页试述乙酰胆碱生物合成主要底物、以及影响乙酰胆碱生物合成主要原因。怎样调整乙酰胆碱合成?Question

222乙酰胆碱和其他神经递质第22页ATP--vesicleprotein---Ach+VesicleAcetylcholineTransporter(VAChT)(II)StorageandreleaseStorage:胆碱能神经元标志高浓度质子泵23乙酰胆碱和其他神经递质第23页Release:PhosphorylationanddephosphorylationofsynapsinIinregulationofreleasableandreservepoolsofvesicles.(II)StorageandreleaseCa++synapsinIactinActivezoneofsynapse.24稳定池不稳定池盈余池乙酰胆碱和其他神经递质第24页Electrophysiologicalobservations:-restingneuromuscularjunctionsoftenhadspontaneousminiatureendplatepotentials(mepps)oflessthan1mVinamplitude.微小（最小）终板电位amplitudeofstimulatedeppweremultiplesofthemepps.Conclusion:-transmitterisreleasedindiscreteunits(quanta).-smallestmepprepresentsthereleaseof1quanta.(II)StorageandreleaseEvidenceofquatarelease:25乙酰胆碱和其他神经递质第25页2.ElectronmicroscopyEvidenceofquatarelease:Freeze-fractureelectronmicrograph26乙酰胆碱和其他神经递质第26页3.NeurochemistryNa+K+Ca2+VSCCreleaseeffluxLactatedehydrogenase*Evidenceofquatarelease:4.CloningofVAChTandfindingitsblockervesamicolQuestion3什么是量子释放？依你所知，有哪些证据证实神经递质是以量子方式释放？检测-释放量27乙酰胆碱和其他神经递质第27页SNAP-25SYNTAXINSynaptobrevin(VAMP)Botulinumtoxin肉毒杆菌毒素Tetanus

toxin破伤风毒素-bungarotoxin-bungarotoxinH+vesamicol银环蛇毒素α-BGT与运动终板乙酰胆碱受体结合，抑制乙酰胆碱对横纹肌细胞膜去极化作用，横纹肌松弛。β-BGT作用于运动神经突触前膜产生一个三相改变，1）递质释放降低，2）释放增加，3）随即是深入抑制，使骨骼肌不能兴奋收缩而转入连续性麻痹，其毒性比突触后毒素高得多。运动终板Blackwidowspidervenom1）阻断SNARE蛋白分离2）形成离子通道28乙酰胆碱和其他神经递质第28页TransmitterisGLYCINETransmitterisACETYLCHOLINEBotulinumToxin29乙酰胆碱和其他神经递质第29页Specificcholinesterase(AchE)：mainlyinneuraltissues。Non-Specificcholinesterase(butyrylcholinesterase):gliacells,non-neuraltissues.I.Enzymaticdecompositionandinactivation乙酰胆碱酯酶-水解速度106Ach/S丁酰胆碱酯酶SectionIII:Enzymaticdecompositionandinactivation30StructureofAchE1)Anionsite:imidazolegroupofhistidine(组氨酸）.2)SiteofEnzymaticdecomposition:-OHofserine,nucleophilic乙酰胆碱和其他神经递质第30页（I）reversiblePhysostigmine,毒扁豆碱neostigmine:新斯明

为含季铵基团氨基甲酸酯类。Edrophonium:Edrophoniumchloride依酚氯铵：

含季铵基团，但无氨基甲酰基。起效快，作用短。（II）irreversible:organicphosphoruspesticide,chemicalwarfareagent（VX,沙林）II.Cholinesteraseinhibitors31乙酰胆碱和其他神经递质第31页有机磷农药32乙酰胆碱和其他神经递质第32页生化武器33乙酰胆碱和其他神经递质第33页antidotestoanticholineesterasessymptomatic“antidote”-atropinepralidoxime(2-PAM)解磷定BACD解毒剂A氯磷定(2-PAM·Cl)

B双复磷(LüH-6)

C甲磺磷定(P2S)

D双磷定(TMB-4)1.2.直接与有机磷结合34解毒：乙酰胆碱和其他神经递质第34页Alzheimer’sdisease四氢氨基吖啶（tetrohydroaminoacridine）

TacrineHydrochloride35抑制胆碱酯酶药用价值乙酰胆碱和其他神经递质第35页SectionIV:CholinergicReceptorsAch一些效应可被毒蕈碱（muscarine）模拟而被阿托品阻断另一些效应可被烟碱（nicotine）模拟而被箭毒（curare）阻断36乙酰胆碱和其他神经递质第36页I.Muscarinicreceptor1.G蛋白偶联2.7次跨膜3.差异--大环4.两类M1,3,5;M2,437乙酰胆碱和其他神经递质第37页Meffects:

1)Inhibitingheart(M2)todecreaseheartrate,conductionandcontractility.

2)Vasodilationofartery(M1血管内皮--

NO）

3)Contractionofbronchial,gastrointestinalandgenitourinarysmoothmuscles,butrelaxationofsphincters

4)Increasing

secretion(M3)ofsalivary,respiratoryandgastrointestinaltractglands

5)Eye(M3):miosis.nearvision.泌尿生殖器括约肌瞳孔缩小38外周效应乙酰胆碱和其他神经递质第38页M1，3，4receptors----cortex,hippocampus—learningandmemoryM1,4receptors----striatum–extrapyramidalmotorcircuitsM2–basalforebrain—autoreceptorstocontrolAChsynthesisandreleaseM5istheleastabundant—Substantianigra.Mreceptorantagonists(trihexyhenidylandbenztropine)areprescribedtoPDpatients苯海索苯托品Tricyclicantidepressantsandlow-potencyantipsychoticdrug(chlorpromazine氯丙嗪)antagonizeMreceptors中枢M受体分布东莨菪硷M1andM2MuscarinicReceptorSubtypesRegulateAntidepressant-LikeEffectsoftheRapidlyActingAntidepressantScopolamineJPharmacolExpTher351:448–456,39乙酰胆碱和其他神经递质第39页II.Nicotinicreceptor40乙酰胆碱和其他神经递质第40页Purificationofcholinergicreceptors

金环蛇Bungarusfasciatus银环蛇Bungarusmulticinctus41应用金银环蛇毒素乙酰胆碱和其他神经递质第41页PurificationofreceptorsCarb-nAchRTorpedoelectricorgannAchRnAchRnAchRsolubilizationHomogenizationCarbamylcholineSequenceanalysisofsegmentsSynthesisofprobeRNAa-BTXReconstructioninlipidbilayer(functional?)电鳐42氨基甲酰胆碱洗脱乙酰胆碱和其他神经递质第42页cDNAlibraryscreeningLowdensityreceptorsintissueIsolatetotalRNAsIsolatemRNAsprobeRNAcDNAlibraryInvitrotranscriptionofthecDNAlibraryInjectionofcRNAsintoXenopusoocytesElectrophysiologicalmeasurementofligand-inducedcurrent SequencingInjectionofcRNAsintoXenopusoocytesPCR43HomologyscreeningTorpedoelectricorgan乙酰胆碱和其他神经递质第43页SubunitsofAchRare4TMHydropathyindexanalysis44乙酰胆碱和其他神经递质第44页thesuperfamilyofcys-loopreceptors亲水面形成离子通道内壁45乙酰胆碱和其他神经递质第45页TM2segmentisthepore-liningdomainofAchR46乙酰胆碱和其他神经递质第46页Structureandfunctions：Membrane-bindingglucoprotein.Fivehomologoussubunits(1,

,1,1,)makeupthechannel.Eachsubunitshasfour-helicalmembranespinningdomains(M1-M4),andM2domainofeachsubunitmakeupthelinenofthechannel.ThetwosubunitsarethebindingsitesforACh

(-BGTX)Bothareceptorandachannel,mediatesfastsignaling.(I)Peripheralnicotinicreceptors47乙酰胆碱和其他神经递质第47页Ach功效？48解剖位置属于外周受体分型属于中枢乙酰胆碱和其他神经递质第48页NolossofAPinnervecellsAristotleOnassislossofmuscletoneMG-myastheniagravisAutoimmunedestructionofacetylcholinereceptorsnAChRHinderACh-receptorinteractionIncreasereceptorturnoveranddegradationTreatment:AChEinhibitors,glucocorticoidsorotheragentstosuppressimmunefunction重症肌无力机制49乙酰胆碱和其他神经递质第49页试述外周尼古丁型乙酰胆碱受体结构和功效特点Question450乙酰胆碱和其他神经递质第50页（II）Centralnicotinicreceptors(includingganglion)Subunitcompositionofcentralnicotinicreceptors:2-10(theperipheralsubunitas1)2-4(theperipheralsubunitas1)About2-6,2-4:Donotformhomomericreceptors,noreceptorswithonlyoronlysubunitsexist.51乙酰胆碱和其他神经递质第51页Commonreceptortypes52乙酰胆碱和其他神经递质第52页Theheteromeric

42

23

subtype:facilitateGABAergicandglycinergicneurotransmission.Insensitiveto-BGTX，sensitivetoKappaorFtoxin.GABAglycine

42

23

42

2353乙酰胆碱和其他神经递质第53页Thehomomeric

75subtype:facilitateglutamatergicneurotransmission.sensitiveto-BGTX.glutamate

75

7含有-BGTX高亲和力结合部位

7-10只形成同聚体（成熟神经元细胞表面）54乙酰胆碱和其他神经递质第54页GarduñoJetal.J.Neurosci.;32:15148-15157©bySocietyforNeuroscience例外Presynapticα4β2nicotinicacetylcholinereceptorsincreaseglutamatereleaseandserotoninneuronexcitabilityinthedorsalraphenucleus.JNeurosci.Oct24;32(43):15148-5755乙酰胆碱和其他神经递质第55页Fig.1(A)PhotomicrographshowingbasalocorticalPHA-L-labeledfibersintheretrosplenialgranularcortex(RSG)followingaseriesofinjectionsofPHA-Lintothemedialpartofthehorizontallimbofthediagonalban(HDBm).PHA-Llabeledfibersarecon...Localizationofα7nicotinicacetylcholinereceptorimmunoreactivityonGABAergicinterneuronsinlayersI–IIIoftheratretrosplenialgranularcortexNeuroscience,Volume252,,443-459N受体在细胞体分布56乙酰胆碱和其他神经递质第56页3,5,72,4makeupthechannel.homomericreceptorscontaining3inmostcondition.Heteromericreceptorscontaining7(胚胎期)(III)Nicotinicreceptorsinautonomicganglions解剖分类属于外周型亚单位组成属于中枢57乙酰胆碱和其他神经递质第57页PresynapticnAChRsinducereleaseofanumberofneurotransmittersPostsynapticnAChRsdepolarizeneurons,increasetheirfiringrate,andcancontributetolong-termpotentiation58主要中枢作用乙酰胆碱和其他神经递质第58页兴奋效应直接--兴奋2.间接？钙依赖效应与N受体钙通透激活邻近电压依赖性钙离子通道细胞内钙参加效应。59乙酰胆碱和其他神经递质第59页试述中枢尼古丁型乙酰胆碱受体结构和功效特点Question4Characteristicsofcentralnicotinereceptors:

*Structure，subunits,*highlypermeabletoCa++.consequentlyactivatevoltagegatedCa++channels.*Facilitationofglutamatergicneurotransmissionsby….sensitiveto….

*FacilitationofGABAergicandglycinergicneurotransmission

by…

Insensitiveto…..

60乙酰胆碱和其他神经递质第60页2----Thereinforcingandcognitionenhancingeffects7-----Sensorygating4-----AnalgesiceffectsnAChR中枢功效61？乙酰胆碱和其他神经递质第61页Summaryofmodulatoryeffectsofacetylcholine(ACh)oncorticalcircuitdynamics.ACh与学习记忆关系62IncreasedAChbooststheprocessingofextrinsicafferentprojections,suppressesintrinsicprojections,andlowersthethresholdfortheinductionofLTP.ReducedAChboostsprocessingofintrinsicprojections,raisesthethresholdfortheinductionofLTPandincreasesthelikelihoodthatLTDisinduced乙酰胆碱和其他神经递质第62页ActivationdynamicsandplasticityinModelofthetafunction.InhibitoryoscillationsconvertdifferencesintheAmountofexcitationreceivedbycellswithinanetworkintoatime(phase)code.Threeexamplecellsareshown,eachreceivingadifferentamountofexcitatoryinput:0.0—thiscellreceivesnoinputandneveractivates,itincursnoLTPorLTD;0.3—thiscellreceivesmoderateinputandactivatesonlyduringthetroughoftheoscillations,itincursLTDasaresult;and0.6—thiscellreceivesstronginputandactivatesduringmorethanhalfoftheoscillation,itincursLTPasaresult.Theta

rhythmismodulatedbycholinergicandGABAergicinputsfromthemedialseptum,andincreasesinacetylcholineareassociatedwiththetaoscillations.Hippocampal

twoseparatefrequencycomponents,acholinergic-independent,movement-relatedthetarhythmat8–9Hz(type1)alowerfrequency6–7Hzcholinergic-dependentcomponent(type2)63乙酰胆碱和其他神经递质第63页RoleofnAChRsindevelopmentThedifferentsubtypesofneuronalnAChRsareknowntobedevelopmentallyexpressedintheratbrainand(particularlyfortheα7subtype)areknowntohaveimportantrolesinthedevelopmentofthehippocampalcircuitry.α7receptorsignaling---thematurationandsurvivalofadult-bornneurons,transformationofGABAergiccurrentsfromexcitatoryinearlydevelopmenttoinhibitorysynapseThecognitivedeficitsassociatedwithADmayberelatedtodysfunctionoftheα7nAChRs.α7nAChRcanundergorapidonsetofdesensitization（channelcloseseveninthecontinuedpresenceofanagonist）.therapeuticdrugsthatpotentiateα7receptorsthroughtheremovalofdesensitization(e.g.,PNU-120596)arecurrentlybeingdevelopedtotreatAlzheimer'sdiseaseandotherneurologicalInthebrain(andinparticularthehippocampus),nicotineorAChactivationoftheα7nAChRblockedthereleaseofproinflammatorycytokines.64胶质细胞？乙酰胆碱和其他神经递质第64页Site1: UptakeofcholineSite2: Biosynthesisofacetylcholine(ChAT)，VesicleAcetylcholineTransporter(VAChT).Site3: ReleaseofAch(Note:Thisisnotagoodsiteforthedesignofcholinergicdrugs).Site4: MetabolismofAChbyacetylcholinesterase(AChE)..Site5: PostsynapticandPresynapticreceptors(maintargetfordrugaction).65乙酰胆碱系统干预？乙酰胆碱能神经元分布？受体结构和功效特点中枢乙酰胆碱功效乙酰胆碱和其他神经递质第65页SectionI:Histamine组胺Recognitionofhistamineasanintercellularmessenger:注射组胺—过敏(allergy)，呼吸窘迫抗组胺药广泛应用于临床抗组胺药品镇静作用—组胺唤醒Arousal

中枢作用？DistributionPeripheral:mastocyte,basicyte.Central:Neuronalhistamineintuberomammillarynucleus(TMN).Non-neuronalhistamine:gliacell,endothelialcell肥大细胞，嗜碱性细胞Histamine不易经过BBB中枢外周独立Chapter12Otherneurotransmitters66？神经营养，神经免疫，脑血流，BBB渗透性调整，等乙酰胆碱和其他神经递质第66页H2-receptorantagonists

asdrugscuringpepticulcerMechanismofactionCompetitivelyblockthehistamine(H2)receptorofacid-producingparietalcells

renderingcellslessresponsivetonotonlyhistaminebutalsotothestimulationofacetylcholineandgastrin.Alsoupto90%inhibitionofvagalstimulatedandgastrinstimulatedacidsecretion.

胃泌激素泌酸细胞FourFDA-approved:Cimetidine西咪替丁（Tagamet®）Ranitidine雷尼替丁（Zantac®）Famotidine法莫替丁（Pepcid®）Nizatidine尼扎替丁（Axid®）Roxatidine罗沙替丁H2-receptorantagonists67乙酰胆碱和其他神经递质第67页1.Histaminetransportandmetabolisminneurons组氨酸脱羧酶，HDC组氨-N-甲基转移酶HNMT单胺氧化酶，醛脱氢酶213456甲基组胺68VMAT-2locatedpostsynapticallyandinglia失活形式HAD抑制剂：

-氟甲基组氨酸，

-FMH乙酰胆碱和其他神经递质第68页2.Histaminergicneurons神经元集中，投射广泛（脑腹侧部为多）突触:dendrites(predominately),cellbody(less)递质共存常见（GABA,P物质）tuberomammillarynucleus(TMN)69乳头体核结节核JCompNeurol.Nov15;518(22):4591–4611.Histidinedecarboxylase(HDC),melanin-concentratinghormone(MCH),andorexin/hypocretins(ORX)乙酰胆碱和其他神经递质第69页70乙酰胆碱和其他神经递质第70页BrainhistamineduringdevelopmentIntherat,theliveristhemainsiteofhistaminesynthesisduringlatestagesoffetaldevelopment,althoughanextensive,transienthistamine-containingneuronalsystemcanbefoundintherapheareaofthebrainduringthisperiod.Thissystemisfirstdetectableonembryonicday14(E14),andatE16,histamineandserotoninshowmarkedcoexistenceintheraphearea.ByE18,histaminelevelsarealreadydiminishedinneuronsoftherapheregion.Histamine-containingmastcellspopulatetheratbrainatE18–E19,andthenumberofthesecellsincreasesuntilpostnatalday4.Therattuberomammillarynucleus(TMN)neuronsarebornbetweenE13andE18,andthatHDC组胺脱羧酶isfirstdetectableintheseneuronsonE18.IntracellularhistamineimmunofluorescencecanbedetectedinTMNneuronsonE2071乙酰胆碱和其他神经递质第71页72ProminentsourcesoftheafferentprojectionstoTMtheinfralimbiccortex,lateralseptumandpreopticnucleus.adrenergiccellgroupsC1–C3,noradrenergicgroupsA1–A3,andfromserotonergicgroupsB5–B9,Themainterminalareasofthehistaminergicprojectionsarealsoslightlydifferentindifferentspecies,buttheycoveressentiallyallareasoftheCNS.下边缘皮质:regionintheventromedialprefrontalcortex乙酰胆碱和其他神经递质第72页2.Histaminergicneurons4)Firingproperties:Spontaneouslyactive:pace-makerproperty.

迟缓去极化—动作电位

自发放电机制:persistentsodiumcurrent，voltage-gatedcalciumcurrent.b.Lowfiringfrequency放电频率低:LongerAPduration动作电位时程长.Obviousafterhyperpolariztion(AHP):后超极化显著

机制：voltage-gatedKcurrent,calcium-dependentKcurrent.e.H3autoreceptorsregulatethefiringrate:inhibitionofVDCCs.I-clampV-clampH3激动剂抑制效应乙酰胆碱和其他神经递质第73页(H3receptorantagonist)噻普酰胺<组胺H3受体拮抗药>反向激动乙酰胆碱和其他神经递质第74页组氨能神经元分布电活动特点和机制.Question75乙酰胆碱和其他神经递质第75页3.Histaminergicreceptors组胺受体H1-4,为G蛋白偶联受体。H1-3分布于CNS。H1，H2兴奋;H3抑制(H3多亚型，Splicing)H4肥大细胞，白细胞，骨髓组织。

原位杂交BindingH1H2H3乙酰胆碱和其他神经递质第76页磷脂酰二肌醇H1-Gq/11磷脂酶C

三磷酸肌醇，甘油二酯

抑制钾电导H2—Gs腺苷酸环化酶H3—Gi/o抑制腺苷酸环化酶

抑制钙离子通道

本身抑制，抑制其它递质释放

组胺受体信号通路乙酰胆碱和其他神经递质第77页H1receptors.DepolarizationofaneuronofthepontinereticularformationthroughblockofaK+conductance.H3receptors.Decreaseoffieldexcitatorypostsynapticpotentialinthedentategyrusofthehippocampusevokedbyperforantpathstimulation.Histaminereducedthereleaseofglutamateinthispathway.乙酰胆碱和其他神经递质第78页H2receptors.Blockofthelong-lastingafterhyperpolarizationafterCa2+-inflow(calcium-dependentK+conductance)andblockoftheaccommodationofactionpotentialfiringinahumanhippocampalpyramidalcell.79乙酰胆碱和其他神经递质第79页InteractionsofthehistamineH3receptorwithdopaminereceptors80乙酰胆碱和其他神经递质第80页4.HistamineandthefunctionoftheCNS1)ArousalwakefulnessH1受体相关证据：1.组胺能神经元活动随睡眠觉醒周期波动2.组胺释放有昼夜节律3.损毁或抑制TM神经元造成嗜睡4.脑内注射组胺或H1受体激动剂可延长清醒，缩短睡眠。H1拮抗剂阻断这些效应。

H3拮抗剂有利觉醒81乙酰胆碱和其他神经递质第81页ThehistaminergicTMNnetworkisanessentialcomponentofthesleep–wakeregulatorysystemVentrolateralpreopticnucleus82GABA乙酰胆碱和其他神经递质第82页4.HistamineandthefunctionoftheCNS脱水stressVasopressinreleaseHistamine

-FMH,H3agonists,postsynaptichistmineantagonists(-)2)MaintenanceofendohomeostasisWaterbalance:H1receptor→vasopressinFeeding下丘脑腹内侧核（饱中枢）

三叉神经核（咀嚼控制）

对leptin功效含有permissiveactionThermalregulation（双向改变，先降后升）

下丘脑视前区H1降低体温调定点下丘脑后部H2散热Cardiovascularregulation

升血压，心率减慢

H2受体-动脉压力感受器反射

兴奋交感神经，释放儿茶酚胺；促释放AVP83乙酰胆碱和其他神经递质第83页3)Learningandmemory:peoplepredisposedtoallergysmarter?4.HistamineandthefunctionoftheCNS4)Emotionandanxiety5)Painmodulation脑内组胺参加焦虑情绪形成外周介导伤害性感受器，介导痛觉在中枢起镇痛作用脑内组胺系统可能是对威胁做出反应系统脑内组胺系统兴奋可激活交感系统，释放应激相关激素。组胺参加神经系统疾病机制。84乙酰胆碱和其他神经递质第84页6）Localizationofhistaminereceptorsindifferenttypesofcellsinthebrainthatarepotentiallyinvolvedinblood–brainbarrierpermeabilityandmultiplesclerosis.Hreceptorsinbrainneurons,astrocytes,endothelialcellsandimmunocompetentcellsformacomplexsystemattheinterfaceofbraincapillariesandtheneuropile.Brainendothelialcellsexpressallfourtypesofhistaminereceptors.Astrocytes,whichsurroundbraincapillarieswiththeirendfeet,expresshistamineH1receptors(H1Rs)andH2Rs.HistaminergicandotherneuronswithterminalsaroundtheastrocyteendfeetcontainpresynapticH3Rs.CirculatingactivatedTlymphocytesexpressatleastH1Rs,andpossiblyalsoH2RsandH4Rs.Mastcells,whichexpressH1Rs,H2RsandH4Rs197,arefoundonlyinlimitedperivascularareasoftheratbrain(medianeminenceandthalamus)andarepotentiallyimportantinthehumanbrainandapossiblesourceofH4Rs.85乙酰胆碱和其他神经递质第85页SectionII:nitricoxideRobertFFurchgott,born1916Dept.ofPharmacology,SUNYHealthScienceCenterNewYorkLouisJIgnarro,born1941Dept.ofMolecularandMedicalPharmacologyUCLASchoolofMedicineLosAngelesFeridMurad,born1936Dept.ofIntegrativeBiologyPharmacologyandPhysiology,UniversityofTexasMedicalSchool,HoustondemonstratedthattherelaxationofbloodvesselsdescribedbyFurchgottwasduetoasubstanceproducedintheendothelium,whichhisgroupcalledendothelium-derivedrelaxingfactor(EDRF).In1987,IgnarrodemonstratedthatEDRFwasnitricoxide.86FurchgottandZawadzkishowedthattherelaxationofbloodvesselsdependedonanintactendothelium.showedthatnitroglycerinecausedbloodvesselstorelax,andthatthiswascausedbythereleaseofnitricoxidegas乙酰胆碱和其他神经递质第86页1.Biosynthesisandenzyme底物：L-精氨酸,

O2,NADPH产物：胍氨酸，NO酶：nitricoxidesynthase,NOS.NO合酶NO是极不稳定脂溶性小分子，可快速扩散Whenexposedtooxygen,NOisconvertedintoNO2.2NO+O2→2NO2Inwater,NOreactwithoxygenandwatertoformHNO2.4NO+O2+2H2O→4HNO2血液中血红蛋白是NO主要去除剂（scavenger），也是NO储存和运输工具87乙酰胆碱和其他神经递质第87页NameGene(s)DescriptionNeuronalNOS(nNOSorNOS1)NOS1ProducesNOinneuronaltissueinboththecentralandperipheralnervoussystem.NeuronalNOSalsoperformsaroleincellcommunicationandisassociatedwithplasmamembranes.InducibleNOS(iNOSorNOS2)NOS2A,NOS2B,NOS2CCanbefoundintheimmunesystembutisalsofoundinthebrainandcardiovascularsystem.EndothelialNOS(eNOSorNOS3orConstitutive/cNOS)mNOS

NOS3GeneratesNOinbloodvesselsandisinvolvedwithregulatingvascularfunction.AconstitutiveCa2+dependentNOSprovidesabasalreleaseofNO.eNOSisassociatedwithplasmamembranessurroundingcells.?88nNOS,eNOS是Ca2+-钙调蛋白依赖性酶，为基本型

cNOS(constitutiveNOS)iNOS不依赖Ca2+-钙调蛋白乙酰胆碱和其他神经递质第88页2.Cellactions无储存囊泡扩散传递作用机制：1.激活鸟苷酸环化酶（guanylylcyclase,GC）2.修饰蛋白质含铁辅基，巯基，和芳香族氨基酸—参加炎症和细胞损伤等病理过程第二信使激活蛋白激酶cGMP-dependentproteinkinase(PKG)additionalsecondmessengersystemscAMP-dependentkinasePKA89乙酰胆碱和其他神经递质第89页注意：它能够扩散到其它细胞90效应：促进突触功效，促进LTP乙酰胆碱和其他神经递质第90页3.FunctionsofNOinCNS（1）Modulationofneurotransmissionexertslong-lastingeffectsthroughregulationoftranscriptionfactorsandmodulationofgeneexpression（2）Leaningandmemory:

modulationofsynapticactivity,neuralplasticity，LTPandmemoryfunctions.（3）RegulationofneuronalsurvivalanddifferentiationNOdonors含有促进记忆。调整LTP，LTDNOdonorsareinuseinclinicalmanagementofcardiovasculardiseases.AmongNOdonors,molsidomine(脉导敏)hasahighbioavailability,along-lastingdurationofaction,likelycrossesthebloodbrainbarrier(BBB)91乙酰胆碱和其他神经递质第91页92亚硝基化92乙酰胆碱和其他神经递质第92页NO在神经系统主要作用机制QuestionWithlowconcentrationsNOisneuroprotectiveandmediatephysiologicalsignaling(e.g.,neurotransmissionorvasodilatation),whereashigherconcentrationsmediateimmune/inflammatoryactions