还剩17页未读,继续阅读
本资源只提供10页预览,全部文档请下载后查看!喜欢就下载吧,查找使用更方便
文本内容:
大鼠行为学实验评价汇总大鼠行为学评定方法比较大鼠进行行为学评定的十分重要,对其评定的方法也颇多,behaviortest但究竟那种方法更适用,目前未有人进行过比较为了合理选择MCAO后的评定方法,如下对目前常用的几种行为学评定方法进行比较行为学检查方法:由位参加试验的人员分别以单盲法对试验的大鼠进3行打分和记录.然后将组的记分结果进行平均后的得分进行统计计算3国内由一个对试验实施过程不了解的观察者对大鼠进行行为学评测评测续贯进行如果大鼠在一次评测中出现恰当的行为,而以后却未出现,按前者记分国外神经行为学检查评分法
1.Longa评分法姿势反射测验
2.Berderson posturalreflex test攀绳实验3,网屏测验
4.screen test肢体放置测验
5.limb-placement testElicited ForelimbPlacing开野试验法测定行为
6.Open—Field
7.MNSS.转动杆测验8rotating poletest肢体对称试验评分法
10.
15.Forepaw OutstretchingThe rat wasbrought up to theedge of the tableand madeto walk on forelimbswhilebeing heldby the tail.Symmetry in the outstretchingof both forelimbswas observedwhile the rat reachedthe tableand the hindlimbs werekept intheair.Scores indicatethe following:3,bothforelimbswere outstretched,andthe ratwalked symmetricallyon forepaws;2,left sideoutstretched less thanthe right,and forepawwalking wasimpaired;1,left forelimbmovedminimally;and0,left forelimbdid not move.Julio H.Garcia,MD;Simone Wagner,MD;Kai-Feng Liu,MD;Xiao-jiangHu,MD,PhD NeurologicalDeficit andExtent ofNeuronal NecrosisAttributable to MiddleCerebral ArteryOcclusion inRats oStroke.1995;26:627-
63516.ClimbingoThe rat was placed on thewall ofa wirecage.Normally the rat usesall fourlimbsto climbup thewall.When the rat wasremoved from the wirecage bypullingit offby thetail,the strengthof attachmentwas noted.Scores indicatethe following:3,rat climbedeasily andgripped tightlyto thewire;2,left sidewasimpaired whileclimbing ordid notgrip ashard asthe rightside;and1,ratfailed toclimb ortended tocircle instead of climbing
17.Body ProprioceptionThe ratwastouched with a blunt stick oneach sideof thebody,and thereactionto thestimulus wasobserved.Scores indicatethe following:3,ratreacted by turning headand wasequally startledby thestimulus onboth sides;2,rat reactedslowly to stimulus on left side;and1,rat did not respondto thestimulusplacedon the left side.
18.Response toVibrissae TouchAbluntstickwas brushedagainst thevibrissae oneach side;the stickwasmoved towardthe whiskersfrom therear of the animal to avoidentering thevisualfields.Scores indicatethe following:3,rat reactedbyturninghead orwasequally startledby thestimulus onboth sides;2,rat reactedslowly tostimulusonleft side;and1,rat did not respondtostimuluson theleft side.改善记忆作用.跳台试验1跳台装置为的被动条件反射箱,四周用黑色塑料lOcm X15cm X60cm板、底面铺可通电的铜栅,每间内放置一高直径的橡皮
4.5cm,
4.5cm垫作为动物回避电击的安全区,先将动物放在反应箱内的橡皮台上适应然后立即通以交流电,动物受到电击后逃避反应为跳上橡3min,40V皮台记录内大鼠受到电击次数,称为错误次数后在底部铜5min24h栅通电情况下,直接将动物置于平台上,记录第次跳下的潜伏期和15内的错误次数min避暗实验
2.Avoidance response根据大鼠有嗜暗习性分设明暗两室明室上方处悬一个日20cm60W光灯,在暗室的中后部分底部铜栅加电压,两室之间设有一直径40V为大小的圆洞实验时将大鼠背对洞口放入明室,同时启动计时3cm表,动物自放入明室至进入暗室遭电击所需时间为潜伏期,鼠受电击后可自行逃出暗室记录潜伏期及内动物受电击次数,后复5min24h测.穿梭箱试验3水迷宫试验
4.Morris水迷宫为圆形水池.直径深水深水温
4.1Morris100cm.50cm.30cm.24水而覆一层塑料泡沫屑.池壁上个等距离点分水池为个象限.±2℃.44在第三象限中央放置平台.平台无色透明.直径高平台没于6cm.28cm.水而下水池周围参照物保持不变2cm.试验包括:
①定位航行试验实验历时将大鼠从place navigationtest:7d入水面向池壁置入水槽中.记录大鼠从入水到爬上平台所需的时间.即逃避潜伏期每只大鼠每人从个不同象限入水点入水进行训练各次.41取其平均值为其每人逃避潜伏期训练中.若大鼠在内找到平台.60s让其于平台上站立若未找到用棒将其引上平台.并让其站立潜10s;10s.伏期限记为将大鼠从平台上拿卜来休息之后.再按序由下一60s60s入水点入水进行下一次实验
②空间探索试验第天spatial probetest8撤去平台.将大鼠从第二象限入水点放入水槽.记录内其在平台象60s限的滞留时间水迷宫试验主要用于测试大鼠对空间位置觉和方位觉的学习记忆
4.2能力水迷宫试验测试的一个重要指标是大鼠的平均逃避潜伏期”
[5]”平均逃避潜伏期指的是大鼠在放入迷宫后学会在中内找到平台所需5要的平均时间,主要与海马功能有关,与大脑皮层也有一定的关系Morris WaterMazeThis is a taskthat measureslearning andmemory in an aqueousenvironment.Briefly,the ratwas placed in a4-ft-diameter watertank thatis visuallyseparatedinto fourquadrants.In thecenter ofone quadrant,the goalquadrant,a platformis hidden1inch belowthe waterline.The ratwas placedin thetankat oneof fourstart locationsnorth,south,east,west andswam forupto60seconds untilit foundthe platform.Once the animal locatedthe platform,itremained onit for20seconds.In theevent that an animaldid notlocate theplatform,the observerpositioned the animal on the platform at theend of the60-second swimperiod.During a2-to4-minute intertrialinterval,theanimals werekept warmby infraredheating lamps.The temperatureof thewatermaze was20±
0.5℃.The latencyand pathlength of theratweremeasured bya videotracking system interfaced toa computer.Nancy E.Alexis,MS;W.Dalton Dietrich,PhD;Edward J.Green,PhD;Ricardo Prado,MD;Brant D.Watson,PhD.Nonocclusive CommonCarotidArtery Thrombosisin theRat Resultsin ReversibleSensorimotor andCognitiveBehavioral Deficits.Stroke.1995;26:2338-
2346.迷宫学习记忆训练及测试:型电迷宫臂长高宽
4.3Y Y30cm.15cm.每臂末端有信号灯示安全区.箱底铜栅间隔电流强15cm15W4mm度选用电压电击延时设一臂为起步区.按臂
0.7mA.38V2s I-II-HI-I顺序轮流作为安全区大鼠在起步区静置予以电击致其逃至安3min.全区.灯光持续然后熄灯体急开始下一次操作15s.45s.学习训练及测试:规定大鼠受电击后从起步区直接逃至安全区为“正确反应”.以达到连续次电击均为正确反应即前所需的电击次1010/10数即尝试次数表示其学习获得能力若尝试次数超过acquisition30次则不再测试.并以次为最大值计数30记忆再现测试:选上述达到标准的大鼠.休息后再次retrieval10/1024h同法测试.以达标准前的尝试次数表示记忆再现能力10/10Motor behavior1observation ofspontaneous ipsilateralcircling,graded from0no circlingto3continuous circling;2contralateral hindlimbretraction,which measuredthe abilityof theanimal to replacethehindlimbafter itwas displacedlaterally by2to3cm,graded from0immediate replacementto3replacement afterminutes ornoreplacement;3beam walkingability,graded0for a rat thatreadily traversesa
2.4-cm-wide,80-cm-long beamto3for a rat unableto stayon the beam for10seconds;平衡木测验balance beamtest平衡木长宽的方木棒,平放在距离地面处,让大鼠80cm,
2.5cm10cm在其上行走评分标准为.个等级分:能跳上平衡木,在上面行走60不会跌倒;分:能跳上平衡木,在上面行走跌倒机会小于分T50%2;能跳上平衡木,在上面行走跌倒机会大于分在健侧后肢帮50%3;助下能跳上平衡木,但受累瘫痪侧后肢不能帮助向前移动;分在平4衡木上不能行走,但可坐在上面;分将大鼠放在平衡木上会掉下来5Beam balanceperformance ofanimals was assessed on a6-point scale17:二二0=balances withsteady posture;1grasps sideof beam;2hugs beam and1limb,falls downfrom beam;3=hugs beam and2limbs,falls downfrombeam,or spinson beam60seconds;4=attempts to balance onbeam butfallsoff40seconds;5=attempts to balance onbeam butfalls off20seconds;and6=falls off,no attempttobalanceor hangonto beam20secondsThis isa test of sensorimotor integration specifically examining hindlimbfunction.The testing apparatus andrating procedureswere adaptedfromFeeney andcolleagues.16A1-inch-wide beam,4ft inlength,is suspended3ft abovethe floorin adimly litroom.At thefar endof the beam isa darkenedgoal box with a narrowentryway.At equaldistances alongthe beam,four3-inch metalscrews arepositioned,angling awayfrom the beams center.Awhite noisegenerator andbright lightsource atthe startof thebeam motivatetheanimaltotraverse thebeamandenter thegoal box.Once insidethe goalbox,the stimuliare terminated.The ratslatency toreach thegoal boxinseconds andhindlimb performance as hetraversed thebeam basedon a1to7rating scaleare recorded.A score of7indicates normalbeam walkingwithless than2foot slips,and ascore of1indicates thattheratis unableto traversethebeam inless than80seconds.Each ratis trained for3days beforesurgeryto acquirethe taskand toachieve normalperformance ascore of7on threeconsecutivetrials.Three baselinetrials arecollected24hours beforesurgery,and threetesting trials are recordeddaily thereafter.Mean valuesof latencyandscore foreach daywere computed.Beam BalanceBeambalance issensitive tomotor corticalinsults.1617This taskis used toassess grossvestibulomotor functionby requiringarattobalancesteadily ona narrow beam.The testinvolves three60-second trainingtrials24hoursbefore surgeryto acquirebaseline data.The apparatusconsists ofa3/4-inch-wide beam,10inches inlength,suspended1ft abovea tabletop.Therat ispositioned on thebeamand mustmaintain steadyposture withall limbsontop of thebeamfor60seconds.The animafsperformance wasrated withthescale ofClifton andcolleagues,18which rangesfrom1to6,withascoreof1being normaland ascore of6indicating thattheanimal was unabletosupport itselfonthebeam.Beam WalkingThisisatestofsensorimotorintegrationspecificallyexamininghindlimbfunction.The testingapparatus andrating procedureswere adaptedfromFeeney andcolleagues.16A1-inch-wide beam,4ft inlength,is suspended3ft abovethe floorin adimly litroom.At thefar endof thebeam isa darkenedgoal box withanarrowentryway.At equaldistances alongthebeam,four3-inch metalscrews arepositioned,angling awayfromthebeams center.Awhite noisegenerator andbright lightsource atthe startof thebeam motivatetheanimaltotraverse thebeamandenter thegoalbox.Once insidethe goalbox,the stimuliare terminated.The ratslatency toreach thegoalboxinseconds andhindlimb performanceas hetraversed thebeam basedona1to7rating scaleare recorded.A score of7indicates normalbeam walkingwithless than2foot slips,and ascoreof1indicates thattheratis unableto traversethebeam inlessthan80seconds.Each ratis trainedfor3days beforesurgeryto acquirethe taskand toachieve normalperformanceascoreof7on threeconsecutivetrials.Three baselinetrialsarecollected24hours beforesurgery,and threetesting trialsare recordeddaily thereafter.Mean valuesof latencyandscore foreach daywere computed.4bilateral forepawgrasp,which measuresthe ability to holdonto a2-mm-diameter steelrod,graded0for arat withnormal forepawgraspingbehavior to3for arat unableto graspwith theforepaws.The scoresfrom all4tests,which weredone overa periodof about15minutes oneach assessmentday,were addedto givea motordeficit scoremaximum possiblescore,
12.Skilled forelimbfunctionlstaircase feedingapparatusThis requiredpretraining beforeinduction ofICH.Rats hadfree access tofood andwater duringfirst2days afterarrival fromthe supplier.The ratswerehoused inpairs instandard plasticboxes witha12-hour day/night cycle.During the following7days,the rats were fed8to15g/dofstandardlaboratory chow to reducetheir body weight to85%to90%oftheinitialweight.Hunger wasthe incentiveto performin the testingapparatus.Thestaircase pretrainingwas performedtwice perday,withatime intervalof atleast4hours betweentrials.Theratwas placedin aclear plasticbox withafood-baited staircaseon eitherside.Each staircasehad7steps,each withawell containing345-mg pelletsP.J.Noyes CoInc.The number of foodpelletsreached andeaten in20minutes was counted.When aplateau wasreached,the topwell wasno longerbaited withpellets,because thesecan bereachedwith thetongue.An additional4to6trials were usedtocalculatemean pretrainingnumberofpellets eatenfrom each side.If the side-to-sidedifference was4onthefinal trials,thesideon whichrat collectedmorepellets wasdesignated itspreferred side.ICH wasinduced in the dominantbrainhemisphere inrats witha preferredside.After pretraining,the ratswereallowed freeaccesstofood for2days beforesurgery andduring the4weeksafter ICH.Beginning28days afterICH,the ratswere fed10to12g/dstandard laboratorychowtodecrease thebodyweightto90%ofthefreefeeding level.They werethen evaluateddaily inthe staircaseapparatus for3weeks.The topwell ofthe staircaseapparatus wasnot baited.The numberoffood pelletseaten in20minutes oneachsidewascountedmaximum possible18per side.nociception
1.plantar testRatswere placedinaclear plasticchamber18x29x13cm,
2.2-mm thickness.Acclimation ofthe animals to theenvironment occurred20minutes beforebaselinetesting for3consecutive dayswith randomactivation ofheatingsource toaccommodate theanimalstothe painstimulus.The heatsource wasthenpositioned underthe glassfloor beneaththe rightor leftforepaw.Pawwithdrawal automaticallyactivated thetimer witha latencyof
0.1second.Measurements weretaken5times at15-minute intervalsand aregiven asaveragedpaw-withdrawal latency.
11.rotarod test
12.adhesive-removal somatosensorytest
13.Spontaneous Activity
14.Symmetry inthe Movementof FourLimbs
15..Forepaw Outstretching
16.Climbing
17.Body Proprioception
18.Response toVibrissae Touch改善记忆作用.跳台试验
1.避暗试验
2.穿梭箱试验
3.水迷宫试验4Motor behavior1observation ofspontaneous ipsilateralcircling,2contralateral hindlimbretraction,平衡木测验3beam walkingability,balance beamtest4bilateral forepawgrasp,Skilled forelimbfunctionlstaircase feedingapparatusnociception1plantar test神经行为学检查
1.评分法Longa神经学检查分个等级5分正常,无神经功能缺损;分左侧前爪不能完全伸展,轻度神经功能缺损;1分行走时,大鼠向左侧(瘫痪侧)转圈,中度神经功能缺损;2分行走时,大鼠身体向左侧(瘫痪侧)倾倒重度神经功能缺损;3分不能自发行走,有意识丧失4评分法姿势反射测验)提尾悬空试验
2.Berderson posturalreflex test(大鼠提尾悬空一米,观察前肢屈曲情况正常大鼠向地面伸展双上肢可伸展双上肢向地面并无其它神经缺陷大鼠得分梗死后大鼠持续屈0曲损伤半球对测患肢;各种不同姿势,从轻度腕屈曲、肩内收、肘伸展到严重姿势,完全腕、肘屈曲,肩内旋转并内收有任何持续前肢屈曲和其它不正常得分大鼠被放于大的软垫子上,表面有大鼠可1用爪子牢牢抓住的塑料压膜纸用手抓其尾巴,缓慢在鼠肩后给予侧压直至前肢滑动几英寸此手法在每个方向重复几次正常或轻度功能损伤大鼠两侧抵抗滑动力相同严重功能损伤大鼠瘫痪侧始终抵抗力下降,得分然后让大鼠自由活动,观察转圈行为大鼠持续向2瘫痪侧转圈得分前肢屈曲带有侧推抵抗力下降经常见到;双上肢3屈曲并侧推抵抗力下降大鼠经常有转圈行为神经检查在分钟内完3-5成正常分未见行为缺陷中度分前肢屈曲(即提尾悬空实验阳性)1重度分侧推抵抗力下降(即侧向推力实验阳性).伴前肢屈曲.无转圈2行为分同级行为.伴自发性旋转32提起鼠尾.使其离地高.观察前肢情况正常大鼠两前肢对称向
0.33m前伸开;如果有肩部内旋、前肢内收现象发生者.根据其严重程度.最高评分为分.正常评分为分将动物置于平滑地板上.分别推其左、右04肩向对侧移动,此时检查动物抵抗推动时的阻力,正常大鼠两侧推动的阻力明显对称如一侧阻力下降者.可根据下降程度评分.正1—3常评分为分观察两前肢肌力将动物两前肢置于一金属网上.然后0轻提起大鼠.检查两前肢肌力正常左右对称.如有一侧前肢肌力下降者.根据下降程度评为分.正常评分为分根据上述行为检查1—30的情况.积分值为一分积分越高说明动物行为障碍越严重此10测验对皮层和纹状体的损伤较敏感攀绳实验3本实验主要检查大鼠双前肢和双后肢的协调配合能力取直径的
1.5cm麻绳从高、长和宽分别为的平台上垂下,手术前训练1m1550cm Id每只大鼠.使之能够爬到绳子的上每个测试日测试次评分的1/33标准如下分.不需任何刺激在内爬上平台;分.不需任何刺激在:010s1内爬上平台;分.刺激次数小于次.在内爬上平台;分.刺激15s2530s3次数小于次.在内爬上平台;分.刺激次数大于次.在内爬560s4560s上平台或是刺激小于次.但爬上平台时间超过分.大鼠不能爬560s;5上平台网屏测验
4.screen test网屏训练:网屏为网带.网眼为网板的左右和50cm X40cm1cm X1cm.上方都用高的木板框边.网屏距地面高度为下方铺以25cm80cm.12cm厚的海绵先将网屏水平放置.将老鼠放在其上.然后缓缓地将其一端抬高.在内将此屏风变成垂直位.保持观察大鼠是否会从网屏上下2s5s.来或用前爪抓握住网屏,从而评价前爪抓握能力及肌力情况网屏实验:评分标准分为个等级分:前爪握住网屏达之久,不会掉下405s来;分:暂时握住网屏,滑落一段距离,但没有掉下来;分:在内掉125s下来;分:网屏转动时,鼠即刻掉下来3肢体放置测验
5.limb-placement testElicitedFore limbPlacing肢体放置于三种独立刺激视觉、触觉、本体感觉以评测运动感觉完整性参照等⑵采用的动物行为测试评分方法具体实K aw amata验有:前肢放置实验,包括1视觉亚实验,即前方刺激----------------实验者将动物握于手中,使其前爪悬空,自桌面上方处向桌面缓慢10c m斜线靠近(此时桌子位于大鼠前方),大鼠正常反应为前肢即刻抓向桌面,损伤大鼠则表现为肢体反应延迟0分--------动物肢体放置反应正常;1分-------反应延迟但不超过2s;2分----------反应延迟且超过2s O侧方刺激,此时桌子位于动物侧方,其余实验方法及评分标准同前方刺激)触觉亚实验,将动物置于桶状凹陷实验器内,使其前爪悬空,2(此时大鼠应该既看不见,也不能用胡须触及桌面,用其前爪背侧轻触桌面,刺激深度仅达皮肤和毛发,动物反应及评分同视觉亚实验,触觉刺激同样分前方及侧方刺激)本体觉亚实验,操作及评分同触觉3(亚实验,仅刺激深度不同,本体觉亚实验给予前爪较大压力,刺激深达肌肉及关节该亚实验只有前方刺激动物前肢放置实验总分范围为分,功能损伤越重,得分越高010〜开野试验法)测定行为
6.Open-Field(方法本实验所用敞箱为正方形,长、宽各,高,底面由面80cm40cm积相等边长的正方形块组成,以动物穿越底面的块数为水平16cm25活动()得分,以直立次数为垂直活动()得分,每只动crossing rearing物共进行次测定,每次73min法是测定抑郁症大鼠行为学的经典方法,分水平运动和垂直Open2Field运动个观察面,以观察得分为评分计算单位,记各组大鼠的水平运2动评分及垂直运动评分Elevated openfield activityis characteristicof hippocampallydamagedanimals19202122and hasbeen observedin animalswith striatal/corticaldamage aswell.23Rats areplacedina novelenvironment3x3ft with3-inch-square gridmarkings onthe base.The numberof timesaratcrosses agridline isrecorded fora5-minute period..肢体对称试验评分法7将大鼠置于网中,网眼大小为当大鼠在网络中行走时,计数
2.3x
2.3cm,在分钟内其前爪漏到网眼中的次数计算公式为脑病变对侧前爪2的错步数一病变同侧前爪的错步数/总步数其分值如为正数表明脑病变对侧功能缺损;如为负数表明脑病变同侧功能缺损
8.mNSS体系的设计原理,评分越低表示功能越健全,分表示功能完全健mNSS0全的大鼠modified NeurologicalSeverity Scores见附件
1.转动杆测验9rotating poletest转棒上行走测评:评分标准分为个等级分:转动过程中.鼠可在棒上40行走;分:转动过程中以上时间.鼠不会掉下来;分:转动开始后.
1.60s2鼠从棒上掉下来;分:转动开始前.鼠从棒上掉下来
310.The detectionwas operatedat1week after surgery byinjecting
0.5mg/kg ofapomorphine intoabdominal cavityfor observationofthechangesin Thebehaviours foronce aweek andcontinuous detectedfor6weeks.Ifconstant right-side rotation occurred andThe numbersof Thecircles rotatedweremore than210r per30minutes,those ratswere consideredas successfulmodels:if norotationoccurredor Thedirection ofrotation wasnot constantorconstant right-side orlell-side rotationbut The speed wasslower than210rper30minutes,those animalswere consideredas unsuccessfulmodels.Inadditional torotation,other companionedabnormal behavioursalso shouldbeobserved,such as,tremor,sluggish,grab,and smellexploring,etc.
11.rotarod testratswere placedon anaccelerating rotarodcylinder,and thetime theanimalsremained onthe rotarodwas measured.Thespeedwas slowlyincreased from4to40rpm within5minutes.A trialended iftheanimalfell offthe rungsorgripped the device andspun aroundfor2consecutive revolutionswithoutattempting towalkonthe rungs.The animalswere trained3days beforeMCAO.The meanduration inseconds onthedevicewas recordedwith3rotarod measurements1day beforesurgery.Motor testdata arepresented aspercentageof meanduration3trials onthe rotarodcompared withtheinternal baselinecontrol beforesurgery.
12.adhesive-removalsomatosensory testsomatosensorydeficit wasmeasured bothbefore andaftersurgery.All ratswerefamiliarized withthetestingenvironment.In theinitial test,2smallpieces ofadhesive-backed paperdots ofequal size,
113.1mm2wereusedasbilateral tactilestimuli occupyingthe distal-radial regiononthewrist ofeachforelimb.Theratwas thenreturned toits cage.The timeto removeeachstimulus fromforelimbs wasrecorded on5trials perday.Individual trialswere separatedby at least5minutes.Before surgery,theanimals weretrainedfor3days.Once theratswereabletoremove thedotswithin10seconds,they weresubjected toMCAO.
13.Spontaneous ActivityTheanimalwasobserved for5minutes inits normalenvironment cage.Therats activitywasassessedby itsabilitytoapproach all four wallsofthe cage.Scores indicatethefollowing:3,rat movedaround,explored theenvironment,and approachedat leastthree wallsofthe cage;2,slightly affectedrat movedaboutinthecage butdidnotapproach allsides andhesitated tomove,although iteventually reachedatleastone upperrim ofthecage;1,severelyaffected ratdidnotrise upat alland barelymoved inthecage;and0,rat didnotmove at all.
14.Symmetry inthe Movementof FourLimbsThe ratwas heldintheair bythetailto observesymmetry inthe movementofthe fourlimbs.Scores indicatethefollowing:3,allfourlimbs extendedsymmetrically;2,limbs onleftsideextended lessor moreslowly thanthoseon theright;1,limbs onleftsideshowed minimalmovement;and0,forelimbon leftside didnotmoveatall.Julio H.Garcia,MD;Simone Wagner,MD;Kai-Feng Liu,MD;Xiao-jiangHu,MD,PhD NeurologicalDeficit andExtent ofNeuronal NecrosisAttributableto MiddleCerebral ArteryOcclusion inRats Stroke.o1995;26:627-635o。