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行为实验方法.docx

萌宠菠菠乐园
2024-10-16 07:05

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2.1物体识别实验检测实验动物的非空间认知能力。Testphasetrial(5min)□ObjectAObjectrecognitiontestThetestapparatuswasawhite,open-fieldplasticbox(60cmx50cmx40cm)placedinasound-prooftestingroom,whichwaslitbyconstantillumination(about40lx)toeliminateshadows.Thediscriminatedobjects(A,BandC)wereidentically-sized(5cmx5cmx5cm),cuboid,woodyblocks.ObjectsAandBwereblack,whereasobjectChadablackand-whitepattern.Eachofthethreeobjectswassufficientlyheavytopreventthemicefrommovingit.TheORTconsistedoftwophases:thesampletrialandthetesttrial(Fig.2A).Inordertoacclimatethemice,eachwasfirstplacedintoanemptyboxfor5min.Subsequently,duringthesampletrialthetwoidenticalobjects(AandB)wereplacedonoppositesides(atadistanceof8cmfromthewallsand34cmfromeachother).Tostarta5-minuteperiodofexploration,eachanimalwasplacedinthemiddleofthetwoobjects.Afterthisperiod,theywerereturnedtotheirhomecagesforanintervalof4h.Inthetesttrial,objectBwasreplacedbyobjectC,whichwasnoveltothemiceanddifferentfromeitherobjectAorB.ThemicewerethenlefttoexploreobjectsAandCfor5min.Theanimals'behavioralperformancesduringthetestswerevideo-tapedandtheexplorationtimeofthesampleandtestphasesforeachofthethreeobjectswasrecordedbyatrainedobserver.2.2物体定位实验检测实验动物的空间认知能力。2.2物体定位实验检测实验动物的空间认知能力。ObjectlocationtestTheapparatususedforthistestwasthesameboxthatwasusedintheORT.Thediscriminatedobjects,DandE,weretwoidenticallysized(5cmdiameterand10cmheight),cylindricalvitreousbottles(Fig.3A).TheperiodofacclimationwasperformedasintheORT.Inthesampletrial,objectsDandEwereplacedintheapparatusasdescribedintheORT.After5minobjectexploration,themicewerereturnedtotheirhomecagefora4-hourinterval.Subsequently,inthetesttrial,objectEwasmovedtoalocationthatwasdiagonallyoppositetoobjectD,andthemousewasleftintheboxfor5minexploration.Theexplorationtimeofboththesampleandtestphaseswasrecorded.ThecriterionusedtoassessexplorationinboththeORTandtheOLTwasthatthenoseofthemousewasorientedtowardstheobjectsatadistanceoflessthan2cm.Periodsinwhichtheratmovedaround,climbedoverorsatontheobjectswerenotrecorded.Topreventolfactorycues,theobjectsandthetestboxwerecleanedwith75%ethanolaftereachtrial.Adiscriminationindex(DI)wascalculatedaccordingtothefollowingequation(Zhaoetal.2007;DeRosaetal.2005):DI=(TnTf)=(Tf+Tn):Here,TnandTfrepresentedthetimespent(duringa5-minobservationperiod)exploringnewandfamiliarobjects(orlocations)respectively.2.3旷场实验检测实验动物的运动能力、探索新事物能力及焦虑程度。Micewillbeplacedfor20mininanarean(50*50*40cm)madeofwoodlaminatedwithlight-greyresinandilluminatedwithwhitelight(50lux).After72hrs,micewillundergothesameproceduretotesttheirlong-termhabituation.Distancemoved,meanvelocity,meandistancetothewallandtimespentinthecenterandattheborderwillbeanalyzedwiththesoftwareasdescribed.Thebehaviorofthemiceduringthefirst5minwillbemeasuredwiththeobserver,Thefollowingparameterswillbeanalyzed:riskassessment,sniffing,rearingoffwall,rearingonwall,jumpingandself-grooming.2.4新笼子新物体实验NewcageNewobject检测实验动物探索新事物能力及焦虑程度。NewcageMicewillbeplacedintoanewcagewithfreshbedding,recordedfor5minandthenleftundisturbedinthenewcage.After24hrsthebehaviorinthenewcage(nowfamiliartothemouse)willberecordedfor5minbeforeand5minafterintroducinganewobjectintothecage.Duringtesting,thecagewillbeilluminatedwithwhitedimlight(5lux).Thefollowingparameterswillbeanalyzedwiththeobserver:sniffingandself-grooming,rearing,diggingandeating/drinking.Forthenewobjecttestadditionalparameterswillbescored:riskassessmenttowardstheobject,sniffatobjectAndrearatobject.2.2.5入侵者实验检测实验动物占据领地的竞争性行为Thisclassicaltestforcompetitivebehaviormeasurestheaggressivenenssofaresidentmousetowardsanunfamiliarmale.Ac57BL/6Jageandbodyweight-matchedunfamiliarmalepreviouslyselectedforlowaggressiveness(intruder)willbeintroducedintothehomecageofthefocalmouse(resident).Thetestwilllast10min.Thefollowingparameterswillbescoredbyatrainedobserverusingthesoftware.Theobserver:socialinvestigation,allo-grooming,chasing,attacking,biting,tailrattling,mounting,rearing,diggingandself-grooming.Numberofbiteswillbecalculateddistinguishingbetweenbitesdirectedtowardsvulnerablepartsofthebody(head,genital,ventralarea)andbitesdirectedtowardsnon-vulnerablepartsofthebody(back,sides,hind-limbs).Thisdistinctionisnecessarytoevaluatetheoffensiveversusdefensivequalityofaggressivebehavior2.6自发选择实验检测实验动物的工作记忆。图2.自发选择实验测试示意图自发选择实验监测工作记忆。动物放在叮”迷宫，“T”迷宫由三个相同尺寸(34x5x30cm),的透明支架组成，两个相对的支架和一个中心支架，相对支架有两个可滑动的控制门。在中心支架有一个门可以将动物限制在盲段。动物通常喜好探访未知的区域，比如实验动物先探访'T”迷宫的左侧支架，下一次它应该首选探访右侧支架，但是前提是它必须记住上一次探访是哪一侧支架，由于选择是自发的，并没有任何惩罚或奖赏机制，所以为自发选择实验。在测试中，大鼠被训练4天，每天测试一次，一次包括14次选择机会。测试前大鼠先被控制在盲段。大鼠可以自由地选择进入相对两个支架中的任何一个。一旦进入一侧支架，另一侧支架就被堵住。等大鼠回到中心支架的盲段时，两侧支架的门都被打开，大鼠又可以开始自由新的选择。统计分析大鼠每次选择不同于上一次一侧支架比例。workingmemoryinvolvestheshort-termmaintenanceofinformationrelecanttocurrentgoalsandhasbeenshowntorequiretheprefrontalcortex.Rodentstenttovisitplacestheyhavenotexploredbefore.Ifamousehasexplored,say,theleftarmofaT-maze,itisexpectedtochosetherightsidethenexttimeitisexposedtothemaze,i.e.alterationisexpectedtooccurinanimalsthatrememberwhichwasthelastvisitedarm.Asthebehaviorisnotreinforcedwithanyrewardorpunishmentitiscalled"spontaneous”.Incasealterationisrewardedby,forinstance,foodpelletplacedinthelastnotvisitedarm,thetestisthencalled"win-shift”sincetheanimalmust"shift”thechoice(from,e.g.theleftarmtotherightone)onceithas"won”thereward.Forthespontaneousalterationtestwewilluseaprotocolnamed"continuous”sincemicearenothandledbetweenconsecutivetrialswithinonesessioninordertoavoidthattheirperformanceisinfluencedbyfactorsrelatedtoanxietyandstress.Themazeconsistofthreesamesizearms(34*5*30cm),twooppositearmsandon