考研復(fù)旦細胞學(xué)姐

2023/3/141Chapter1.

IntroductionCellBiology雙語教材2023/3/1421.1.Whatiscellbiology?1.2.Thecelltheory1.3.Cellisthebasicunitoflife1.4.Diversityofcells1.5.Theprokaryoticcell1.6.Theeukaryoticcell1.7.Moderncellbiology1.8.Thetechnologyofcellbiology1.9.TrainingthescientistsoftomorrowContents2023/3/1431.1.Whatiscellbiology?Theapplicationofmolecularbiologicalapproachestoanunderstandingoflifeatthecellularlevel.Themolecularbasisofcellstructure,cellfunctionandcellinteractionsUnderstandingofwholeorganismscomposedwithcells2023/3/144TheBasisofModernBiologyStudythemoleculeswithincellsmacromoleculesandreactions,investigatedbybiochemiststhefunctionalprocessesdescribedbycellbiologiststhegenecontrolpathwaysidentifiedbymolecularbiologistsandgeneticists.2023/3/145CellBiologyIsinProgressTwogatheringforceswillreshapecellbiology:

Thegenomics,thecompleteDNAsequenceofmanyorganisms,Theproteomics,theknowledgeofallthepossibleshapesandfunctionsthatproteinsemploy.DNAmRNAProtein2023/3/146TheStudythemolecularsimilaritiesanddifferencesbetweencelltypesthemolecularcompositionofcellshowcellsworkduetomolecularsimilaritiesanddifferencescloselyrelatedtogenetics,biochemistry,molecularbiologyanddevelopmentalbiology.2023/3/1471.2.ThecelltheoryThreeindispensabletheoriesuponwhichthescienceofbiologyisbuilt.ThetheoryofevolutionThecelltheoryThetheoryofequilibriumthermodynamics平衡態(tài)熱動力學(xué)

CharlesDarwin

(1809-82)JeanBaptisteLemarck1744～1829J.WillardGibbs1839-1903,TheArchitectofModernEquilibriumThermodynamics.2023/3/1482023/3/149

RobertHooke(18July1635-3March1703)AntonyvanLeeuwenhoek(1632-1723)“偉大的看門人”Hegavethefirstcompletedescriptionsofthebacteria,theprotozoans原生動物(whichhecalledanimalcules),spermatozoa,andstripedmuscle.Healsoobservedtheredbloodcellsinhisdetailedstudyofcapillarycirculation.2023/3/1410HeworkedunderJohannesMüllerandstudiedprimarilycellsinplants.Heobservedthatallplantsseemedtobecomposedofcells,andisthusconsideredtheco-founderofcelltheorytogetherwithSchwann.MatthiasJakob,Schleiden,(1804-1881)

Heidentifiedthecellasthebasicstructureofplantandanimaltissuein1938-1839.TogetherwithMatthiasSchleidenheformulatedthecelltheoryoflife.TheodorSchwann(1810-1882),Germanphysiologistandhistologist2023/3/1411SchleidenandSchwannproposedthecelltheoryin1838.Thecelltheorystatesthat:

Alllifeformsaremadefromoneormorecells.Cellsonlyarisefrompre-existingcells.Thecellisthesmallestformoflife.TheCellTheory(1938)2023/3/1412Allknownlivingthingsaremadeupofcells.Thecellisstructural&functionalunitofalllivingthings.Allcellscomefrompre-existingcellsbydivision.

(SpontaneousGenerationdoesnotoccur).Cellscontainshereditaryinformationwhichispassedfromcelltocellduringcelldivision.Allcellsarebasicallythesameinchemicalcomposition.Allenergyflow(metabolism&biochemistry)oflifeoccurswithincells.

ThemoderntenetsoftheCellTheoryThecelltheoryalsoprovidesuswithanoperationaldefinitionof"life".2023/3/14131.3.CellisthebasicunitoflifeAllcellshavesomerecognizablesimilaritiesPlasmamembrane

Nucleus

Organelles

Ribosomesmitochondrionlysosomes2023/3/1414Alllivingthingsarecomposedofoneormorecells.Cellhasthespecialfeaturesoflivingthing.MovementEvenplantsmove,suchasopeningandclosingpetals（花瓣）,andturningtofacethesun.RespirationBreathing.Animalsbreatheinoxygen,andbreatheoutcarbondioxide.Plantsbreatheincarbondioxide,andbreatheoutoxygen.SensitivitytoenvironmentPlantsaresensitivetolightandgravity(stemsgrowupandrootsgrowdown).Animalsaresensitivetomanythingssuchasnoiseandlight.GrowthDoIneedtoexplainthis?ReproductionBabies!Plantsandanimalscanmakemoreofthemselveswithsimilarlyinheritedfeatures.MetabolismAlllivingthingsneedtoeatfoodandgiveoffwaste,suchasmoisture(eg.sweat)orgoingtothetoilet!2023/3/1415DifferencesbetweenPlantCellandAnimalCellPlantCellAnimalCellProkaryoticcellsandplantcellsbothhavearigidcellwallmadeupofpolysaccharides.Plantcellshavethechloroplastwhichisthesiteofphotosynthesis.Whileanimalcellshavethemitochondrionwhichisthesiteofaerobicrespiration.Thevacuoles,largeemptyappearingareasinthecytoplasm,areusuallyfoundinplantcellswheretheystorewaste.Vesiclesaremuchsmallerthanvacuolesinanimalcells.2023/3/1416OriginoftheCellWheredidlifeoriginateandhow?Extra-terrestrialorOrganicChemicalEvolutionStanleyMiller(b.1930)wasadoctoralstudentworkingwithUreyattheUniversityofChicago,researchingpossibleenvironmentsofearlyEarth.In1953hecombinedtheideasofUreyandOparininashort,simpleexperiment.Theearliestcellsmightbeasmuchas3.8billionyearsold.InorganicOrganicAminoacidsPre-cellularSinglecellMulticellularorganisms2023/3/1417Theinteractionsofthesemoleculeswouldhaveincreasedastheirconcentrationsincreased.Reactionswouldhaveledtothebuildingoflarger,morecomplexmolecules.Apre-cellularlifewouldhavebeganwiththeformationofnucleicacids.Chemicalsmadebythesenucleicacidswouldhaveremainedinproximitytothenucleicacids.Eventuallythepre-cellswouldhavebeenenclosedinalipid-proteinmembrane,whichwouldhaveresultedinthefirstcells.2023/3/1418DNARNAProteinRNAhastheabilitytoactasbothgenesandenzymes.Thispropertycouldofferawayaroundthe"chicken-and-egg"problem.(Genesrequireenzymes;enzymesrequiregenes.)RNAcanbetranscribedintoDNA.FromRNAtolifeandtheRNAworldcouldbetheoriginalpathwaytocells.OriginsofLifeandtheRNAWorld

SidneyAltmanandThomasCechwereawardedtheNobelPrizeforBiochemistryin1989Whichcamefirstintheoriginoflife,DNA,RNAorprotein?2023/3/14191.Thepresenceofenzymesandothercomplexmoleculesessentialtotheprocessesneededbylivingsystems.Miller'sexperimentshowedhowthesecouldpossiblyform.2.Thecapacityforreplicationfromonegenerationtoanother.MostorganismstodayuseDNAasthehereditarymaterial,althoughrecentevidence(ribozymes核酶)suggeststhatRNAmayhavebeenthefirstnucleicacidsystemtohaveformed,referredtotheRNAworld.3.Amembranethatseparatestheinternalchemicalsfromtheexternalchemicalenvironment.Thisalsodelimitsthecellfromnot-cellareas.TheworkofSidneyW.Foxhasproducedproteinoidspheres(蛋白質(zhì)小球),whichwhilenotcells,suggestapossibleroutefromchemicaltocellularlife.ThreeThingsMakeCellDifferentfromNon-cellsystem2023/3/1420MicroscopeIsNeededtoVisualizeCells2023/3/1421Prokaryoteshavenonucleusorothermembrane-boundedcompartments.Theylackdistinctorganelles,althoughsomedohaveinvaginatedmembranestructures.Eukaryoteshaveamembrane-boundednucleusandusuallyhaveothermembrane-boundedcompartmentsororganellesaswell.1.4.Diversityofcells2023/3/1422DifferencesbetweenProkaryotesandEukaryotes(Tobecontinued)ProkaryotesEukaryotesSize1-10microns10-100micronsComplexityUnicellular,rarelysmallclustersorfilamentsSometimesunicellularmoreoftenmulticellularMembraneboundorganellesNone(mesosomeisinfoldingofcytoplasmicmembrane)nuclei,mitochondria,chloroplasts,lysosomes,endoplasmicreticulum,golgi,vacuolesNucleusnoyesChromosomeSingleandcircularUsuallyseveralandlinearIntrons&ExonsoccasionallyfrequentHistonesnoyesPloidyhaploidDiploidMitosis&meiosisabsentpresent2023/3/1423SexualreproductionNoneorunidirectionalfromdonortorecipientUsually,involvesfusionofhaploidgametesRibosomes70s(50s+30ssubunits)80s(60s+40s)incytoplasm(mitochondria&chloroplastshaveprokaryoticribosomes)CytoskeletonabsentMicrotubulesandmicrofilamemtsCellwallUsuallypresentAbsentinanimals,presentinfungi{chitin)andplants(cellulose)MotilitySimple,prokaryoticflagella,glidingmotionComplex“9+2”flagellaorciliawithcentriolesDifferentiationUsuallyabsentCellsdifferentiaetoformtissue&organsEnergymetabolismManydiverspathwaysinvariousbacteriaGlycolysisincytoplasm,krebscycleandETCinmitochondriaEndocytosis&cytoplasmicstreamingabsentpresentOxygenAerobicand/oranaerobicUsuallyaerobicSterols甾酮UsuallyabsentUsedashormonesandinplasmamembrane2023/3/1424Twokingdomsofprokaryotes–Eubacteria真細菌(細菌)Archaebacteria原始細菌(古細菌)Threedifferenttypesofbacteria Bacilli桿菌Cocci球菌 Spiral螺旋菌ProkaryotesPhilipHugenholtz:Exploringprokaryoticdiversityinthegenomicera,GenomeBiol.2002;3(2):reviews2023/3/1425BacteriaMorphologicalDiversityFrom2023/3/1426ScaleBacteriavs.EucaryotesProkaryoticcellstructure2023/3/1427CellwallTeichoicacid磷壁(酸)質(zhì)2023/3/1428PlantcellChloroplast類囊體類囊體堆疊2023/3/1429JessicaL.Green,etal:Spatialscalingofmicrobialeukaryotediversity

Nature

432,747–750(2004);Algae藻類Protista原生生物Fungi真菌類Plantae植物Animalia動物Eukaryotes2023/3/1430Plantcellstructure2023/3/1431Animalcellstructure2023/3/1432GeneticControlofCellDiversityThediversityderivesfrombothvarietyingeneexpressionandfromthepatternsofcellularcontrolforgeneproductbehavior.Celldiversityderivesfrombothevolutionanddevelopmentoflivingthings,EvolutiongenomedifferencesamegenomeDevelopment

withdifferentgeneexpressionDifferentcells2023/3/1433Trillions百萬兆ofcells,200+celltypesOnecell,onecelltype2023/3/1434Metazoan(多細胞動物）PhenotypicComplexityfromselectivegeneexpression2023/3/14351.7.ModerncellbiologyContemporarycellbiology,oftenreferredtoasMolecularCellBiology●Geneticcontrolincells●

Threeflowswithincells1)flowofinformation;2)flowofmass3)flowofenergy.●

SystemsBiologyoftheCell1)Molecularcytomics2)Cellulargenomics3)Cellularproteomics4)Cellularepigenomics5)Insilicocell2023/3/1436Themolecularbasisofgeneticcontrolincells,particularlyineukaryoticcellsisoneofthemostbasicactiveareasofmolecularcellbiology.Geneticcontrolincells?Eachcellhaswholegeneticinformation?

Differentcellhasdifferentgenesexpressionpattern?

Thereareinteractionsamonggenes?

Phenotypeisaresultoftheinteractionbetweengenesandenvironment.2023/3/1437FlowofinheritinformationFlowofenergyMaterialflowswithincellsThreeflowswithincellsNoneofthesethreeflowsoperatesinisolationfromtheothertwo.

Theflowofmassandflowofenergyareextremelycloselylinked,asbiologicalmoleculesarebrokenaparttomakeotherbiologicalmoleculesortoharvestenergy.2023/3/1438FlowofinheritinformationGeneticinformationCentraldogma(中心法則)Chromatinremodeling（染色質(zhì)重塑）Histoneacetylation(組蛋白乙?；?Histonemethylation(組蛋白的甲基化)Histonphosphorylation(組蛋白的磷酸化)DNAmethylation(CpG島甲基化)EpigeneticinformationChromatinbasedgeneregulation2023/3/1439Flowofenergy-metabolism—

Someprocesses(anabolic合成代謝)areenergyconsuming.

Otherprocessesareenergyproducing(catabolic分解代謝).—Energyintermediatesarethemolecules,adenosinetriphosphate(ATP)andadenosinediphosphate(ADP),whichareusedtoallowendergonic吸能的reactionstooccurincells.—Energytransductionhappensinmitochondriaandchloroplasts.2023/3/1440Materialflowswithincells

Simplemoleculesabsorbedbycellsareincorporatedintolargermoleculesand/orusedtomakeothermolecules,calledmaterialtransformation.Metabolicpathwaynetworksinacellinclude:CarbohydratemetabolismAminoacidmetabolismLipidsmetabolismNucleotidemetabolism2023/3/1441SystemsBiologyoftheCellCurrently,biologiststendtostudythecellstructureandbehaviorsastheultimateobjectivetosystemicallyunderstandtheeffectof—

themoleculesonthewholecell,and—

theeffectoftheindividualcellonthewholeorganismaswell.cytomics

2023/3/14422023/3/1443E-cell(虛擬細胞

)

Thecomputationalmodeloflivingcell,reflectingthemolecularnetworkswithinacell.

E-CellAconceptofconstructingvirtualcellsoncomputers.E-CellProjectAninternationalresearchprojectaimingatdevelopingnecessarytheoreticalsupports,technologiesandsoftwareplatformstoallowprecisewholecellsimulation.E-CellSystemAnobject-orientedsoftwaresuiteformodeling,simulation,andanalysisoflargescalecomplexsystemssuchasbiologicalcells2023/3/14442023/3/1445CellModelsforResearchofLifeSciencesCellisthebasicunitoflifeCellhasallthefeaturesoflivingthings3.Cellhaswholeinheritmaterial

4.Cellhaswholeregulationongeneexpression5.Thedevelopmentsofmoderntechniquesinresearches

2023/3/14461.8.ThetechnologyofcellbiologyIsolationtechnologyCellCultureFlowCytometryFunctionalBio-imagingofCellModernlightmicroscopyConfocalmicroscopyElectronmicroscopeLivecellImageanalysisRNAInterferenceTechnologyGeneknockouttechnologyGeneMicroarrayProteinChips2023/3/14471.8.IsolationtechnologyCentrifugationVelocityCentrifugation速度離心isodensityCentrifugation等密度離心Chromatography凝膠過濾層析（排阻層析或分子篩）親和層析離子交換層析：陰離子交換劑（DEAE-纖維素樹脂）陽離子交換劑（CM-纖維素樹脂）

2023/3/1448Modernlightmicroscopy（光學(xué)顯微鏡

◆普通雙筒顯微鏡

(binoularmicroscope)

◆熒光顯微鏡

(fluorescencemicroscope)

◆相差顯微鏡

(phasecontrastmicroscope)◆倒置顯微鏡2023/3/1449FluorescenceMicroscopy2023/3/1450Slide40x4DimagingofspindlemovementsLaser

scanning

confocal

microscope

LSCM激光共焦顯微鏡2023/3/1451Tracingmoleculeinlivingcells2023/3/1452Electronmicroscope(電子顯微鏡)transmissionelectronmicroscope,TEM(透射電鏡）Scanningelectronmicroscope,SEM(掃描電鏡）Scanningtransmissionelectronmicroscope,STEM(掃描透射電鏡）2023/3/1453冰凍蝕刻技術(shù)2023/3/1454TECHNIQUESOF

CYTOCHEMISTRY

2.3.1酶細胞化學(xué)技術(shù)

酶細胞化學(xué)技術(shù)就是通過酶的特異細胞化學(xué)反應(yīng)來顯示酶在細胞內(nèi)的定位。2.3.2免疫細胞化學(xué)技術(shù)

免疫熒光技術(shù)

免疫電鏡技術(shù)2023/3/1455間接成像技術(shù)Scanningtunnelingmicroscope,STM(掃描隧道鏡）Atomicforcemicroscope,AFM(原子力顯微鏡）Twophotonmicroscope

2023/3/1456CellCulturePrimaryculture(原代培養(yǎng)）CelllineCellstrainPlanttissuecultureCellfusionMicromanipulation,核移植克隆技術(shù)Monoclonalantibodytechnology2023/3/1457Monoclonalantibodytechnology小鼠脾（B淋巴細胞）X骨髓瘤細胞細胞融合HAT培養(yǎng)基H：hypoxanthine次黃嘌呤A:aminopterin氨基蝶呤：阻斷核酸合成主通道T:thymine胸腺嘧啶（突變腫瘤細胞：無胸腺嘧啶核苷激酶(TK,合成DNA旁路酶)和磷酸核糖轉(zhuǎn)移酶(HGPRT,合成RNA旁路酶）

2023/3/1458流式細胞術(shù)2023/3/1459細胞分選