1、大鼠行為學(xué)實(shí)驗(yàn)評(píng)價(jià)匯總大鼠行為學(xué)評(píng)定方法比較大鼠進(jìn)行行為學(xué)評(píng)定(behaviortest)的十分重要，對(duì)其評(píng)定的方法也頗多，但究竟那種方法更適用，目前未有人進(jìn)行過(guò)比較。為了合理選擇MCAO后的評(píng)定方法，如下對(duì)目前常用的幾種行為學(xué)評(píng)定方法進(jìn)行比較。行為學(xué)檢查方法:由3位參加試驗(yàn)的人員分別以單盲法對(duì)試驗(yàn)的大鼠進(jìn)行打分和記錄.然后將3組的記分結(jié)果進(jìn)行平均后的得分進(jìn)行統(tǒng)計(jì)計(jì)算。(國(guó)內(nèi))由一個(gè)對(duì)試驗(yàn)實(shí)施過(guò)程不了解的觀察者對(duì)大鼠進(jìn)行行為學(xué)評(píng)測(cè)。評(píng)測(cè)續(xù)貫進(jìn)行。如果大鼠在一次評(píng)測(cè)中出現(xiàn)恰當(dāng)?shù)男袨?，而以后卻未出現(xiàn)，按前者記分。(國(guó)外)神經(jīng)行為學(xué)檢查L(zhǎng)onga評(píng)分法Berderson評(píng)分法姿勢(shì)反射測(cè)驗(yàn)(postu

2、ralreflextest)攀繩實(shí)驗(yàn)網(wǎng)屏測(cè)驗(yàn)(screentest)肢體放置測(cè)驗(yàn)(limb-placementtest)ElicitedForelimbPlacing開(kāi)野試驗(yàn)(Open-Field法)測(cè)定行為MNSS轉(zhuǎn)動(dòng)桿測(cè)驗(yàn)(rotatingpoletest)Rotation肢體對(duì)稱試驗(yàn)評(píng)分法rotarodtestadhesive-removalsomatosensorytest13.SpontaneousActivity14.SymmetryintheMovementofFourLimbsForepawOutstretchingClimbingBodyProprioceptionRespo

3、nsetoVibrissaeTouch改善記憶作用跳臺(tái)試驗(yàn)避暗試驗(yàn)穿梭箱試驗(yàn)水迷宮試驗(yàn)Motorbehaviorobservationofspontaneousipsilateralcircling,contralateralhindlimbretraction,beamwalkingability，平衡木測(cè)驗(yàn)(balancebeamtest)bilateralforepawgrasp,Skilledforelimbfunction(1)staircasefeedingapparatusnociception(1)plantartest1神經(jīng)行為學(xué)檢查:Longa評(píng)分法神經(jīng)學(xué)檢查分5個(gè)等級(jí)：0

4、分：正常，無(wú)神經(jīng)功能缺損；分：左側(cè)前爪不能完全伸展，輕度神經(jīng)功能缺損；分：行走時(shí)，大鼠向左側(cè)（癱瘓側(cè)）轉(zhuǎn)圈，中度神經(jīng)功能缺損；分：行走時(shí)，大鼠身體向左側(cè)（癱瘓側(cè)）傾倒。重度神經(jīng)功能缺損；分：不能自發(fā)行走，有意識(shí)喪失。2.Berderson評(píng)分法姿勢(shì)反射測(cè)驗(yàn)（posturalreflextest）提尾懸空試驗(yàn)大鼠提尾懸空一米，觀察前肢屈曲情況。正常大鼠向地面伸展雙上肢?？缮煺闺p上肢向地面并無(wú)其它神經(jīng)缺陷大鼠得0分。梗死后大鼠持續(xù)屈曲損傷半球?qū)y(cè)患肢；各種不同姿勢(shì)，從輕度腕屈曲、肩內(nèi)收、肘伸展到嚴(yán)重姿勢(shì)，完全腕、肘屈曲，肩內(nèi)旋轉(zhuǎn)并內(nèi)收。有任何持續(xù)前肢屈曲和其它不正常得1分。大鼠被放于大的軟墊子上，

5、表面有大鼠可用爪子牢牢抓住的塑料壓膜紙。用手抓其尾巴，緩慢在鼠肩后給予側(cè)壓直至前肢滑動(dòng)幾英寸。此手法在每個(gè)方向重復(fù)幾次。正?；蜉p度功能損傷大鼠兩側(cè)抵抗滑動(dòng)力相同。嚴(yán)重功能損傷大鼠癱瘓側(cè)始終抵抗力下降，得2分。然后讓大鼠自由活動(dòng)，觀察轉(zhuǎn)圈行為。大鼠持續(xù)向癱瘓側(cè)轉(zhuǎn)圈得3分。前肢屈曲帶有側(cè)推抵抗力下降經(jīng)常見(jiàn)到；雙上肢屈曲并側(cè)推抵抗力下降大鼠經(jīng)常有轉(zhuǎn)圈行為。神經(jīng)檢查在35分鐘內(nèi)完成。正常0分未見(jiàn)行為缺陷中度1分前肢屈曲（即提尾懸空實(shí)驗(yàn)陽(yáng)性）重度2分側(cè)推抵抗力下降（即側(cè)向推力實(shí)驗(yàn)陽(yáng)性）.伴前肢屈曲.無(wú)轉(zhuǎn)圈行為3分同2級(jí)行為.伴自發(fā)性旋轉(zhuǎn)。提起鼠尾使其離地0.33m高觀察前肢情況。正常大鼠兩前肢對(duì)稱向前伸

6、開(kāi);如果有肩部?jī)?nèi)旋、前肢內(nèi)收現(xiàn)象發(fā)生者.根據(jù)其嚴(yán)重程度.最高評(píng)分為4分.正常評(píng)分為0分。將動(dòng)物置于平滑地板上分別推其左、右肩向?qū)?cè)移動(dòng)，此時(shí)檢查動(dòng)物抵抗推動(dòng)時(shí)的阻力，正常大鼠兩側(cè)推動(dòng)的阻力明顯對(duì)稱。如一側(cè)阻力下降者可根據(jù)下降程度評(píng)13分正常評(píng)分為0分。觀察兩前肢肌力。將動(dòng)物兩前肢置于一金屬網(wǎng)上然后輕提起大鼠檢查兩前肢肌力。正常左右對(duì)稱如有一側(cè)前肢肌力下降者根據(jù)下降程度評(píng)為13分正常評(píng)分為0分。根據(jù)上述行為檢查的情況積分值為010分。積分越高說(shuō)明動(dòng)物行為障礙越嚴(yán)重。此測(cè)驗(yàn)對(duì)皮層和紋狀體的損傷較敏感。3攀繩實(shí)驗(yàn):本實(shí)驗(yàn)主要檢查大鼠雙前肢和雙后肢的協(xié)調(diào)配合能力。取直徑1.5cm的麻繩從高1m、長(zhǎng)和寬

7、分別為15、50cm的平臺(tái)上垂下，手術(shù)前Id訓(xùn)練每只大鼠使之能夠爬到繩子的上1/3。每個(gè)測(cè)試日測(cè)試3次。評(píng)分的標(biāo)準(zhǔn)如下:0分.不需任何刺激在10s內(nèi)爬上平臺(tái);1分.不需任何刺激在15s內(nèi)爬上平臺(tái);2分刺激次數(shù)小于5次在30s內(nèi)爬上平臺(tái);3分刺激次數(shù)小于5次.在60s內(nèi)爬上平臺(tái);4分刺激次數(shù)大于5次在60s內(nèi)爬上平臺(tái)或是刺激小于5次但爬上平臺(tái)時(shí)間超過(guò)60s;5分大鼠不能爬上平臺(tái)。4.網(wǎng)屏測(cè)驗(yàn)(screentest)網(wǎng)屏訓(xùn)練:網(wǎng)屏為50cmX40cm網(wǎng)帶.網(wǎng)眼為1cmX1cm.網(wǎng)板的左右和上方都用25cm高的木板框邊.網(wǎng)屏距地面高度為80cm.下方鋪以12cm厚的海綿。先將網(wǎng)屏水平放置.將老鼠放在

8、其上然后緩緩地將其一端抬高在2s內(nèi)將此屏風(fēng)變成垂直位保持5s.觀察大鼠是否會(huì)從網(wǎng)屏上下來(lái)或用前爪抓握住網(wǎng)屏，從而評(píng)價(jià)前爪抓握能力及肌力情況。網(wǎng)屏實(shí)驗(yàn):評(píng)分標(biāo)準(zhǔn)分為4個(gè)等級(jí)。0分:前爪握住網(wǎng)屏達(dá)5s之久，不會(huì)掉下來(lái);1分:暫時(shí)握住網(wǎng)屏，滑落一段距離，但沒(méi)有掉下來(lái);2分:在5s內(nèi)掉下來(lái);3分:網(wǎng)屏轉(zhuǎn)動(dòng)時(shí)，鼠即刻掉下來(lái)。肢體放置測(cè)驗(yàn)(limb-placementtest)ElicitedForelimbPlacing肢體放置于三種獨(dú)立刺激(視覺(jué)、觸覺(jué)、本體感覺(jué))以評(píng)測(cè)運(yùn)動(dòng)感覺(jué)完整性。參照Kawamata等采用的動(dòng)物行為測(cè)試評(píng)分方法。具體實(shí)驗(yàn)有:前肢放置實(shí)驗(yàn)，包括(1)視覺(jué)亞實(shí)驗(yàn),即前方刺激實(shí)驗(yàn)者將動(dòng)

9、物握于手中,使其前爪懸空，自桌面上方10cm處向桌面緩慢斜線靠近(此時(shí)桌子位于大鼠前方),大鼠正常反應(yīng)為前肢即刻抓向桌面,損傷大鼠則表現(xiàn)為肢體反應(yīng)延遲。0分動(dòng)物肢體放置反應(yīng)正常;1分反應(yīng)延遲但不超過(guò)2s;2分反應(yīng)延遲且超過(guò)2s。側(cè)方刺激，此時(shí)桌子位于動(dòng)物側(cè)方,其余實(shí)驗(yàn)方法及評(píng)分標(biāo)準(zhǔn)同前方刺激。(2)觸覺(jué)亞實(shí)驗(yàn),將動(dòng)物置于桶狀凹陷實(shí)驗(yàn)器內(nèi),使其前爪懸空,此時(shí)大鼠應(yīng)該既看不見(jiàn),也不能用胡須觸及桌面,用其前爪背側(cè)輕觸桌面,刺激深度僅達(dá)皮膚和毛發(fā),動(dòng)物反應(yīng)及評(píng)分同視覺(jué)亞實(shí)驗(yàn),觸覺(jué)刺激同樣分前方及側(cè)方刺激。(3)本體覺(jué)亞實(shí)驗(yàn),操作及評(píng)分同觸覺(jué)亞實(shí)驗(yàn),僅刺激深度不同,本體覺(jué)亞實(shí)驗(yàn)給予前爪較大壓力,刺激深達(dá)

10、肌肉及關(guān)節(jié)。該亞實(shí)驗(yàn)只有前方刺激。動(dòng)物前肢放置實(shí)驗(yàn)總分范圍為010分,功能損傷越重，得分越高。開(kāi)野試驗(yàn)(Open-Field法)測(cè)定行為方法：本實(shí)驗(yàn)所用敞箱為正方形，長(zhǎng)、寬各80cm,高40cm，底面由面積相等邊長(zhǎng)16cm的正方形25塊組成，以動(dòng)物穿越底面的塊數(shù)為水平活動(dòng)(crossing)得分，以直立次數(shù)為垂直活動(dòng)(rearing)得分，每只動(dòng)物共進(jìn)行7次測(cè)定，每次3minOpen2Field法是測(cè)定抑郁癥大鼠行為學(xué)的經(jīng)典方法，分水平運(yùn)動(dòng)和垂直運(yùn)動(dòng)2個(gè)觀察面，以觀察得分為評(píng)分計(jì)算單位，記各組大鼠的水平運(yùn)動(dòng)評(píng)分及垂直運(yùn)動(dòng)評(píng)分。E1evatedopenfie1dactivityischaract

11、eristicofhippocampa11ydamagedanima1s19202122andhasbeenobservedinanimalswithstriatal/corticaldamageaswell.23Ratsareplacedinanovelenvironment(3x3ft)with3-inch-squaregridmarkingsonthebase.Thenumberoftimesaratcrossesagridlineisrecordedfora5-minuteperiod.肢體對(duì)稱試驗(yàn)評(píng)分法將大鼠置于網(wǎng)中，網(wǎng)眼大小為2.3x2.3cm,當(dāng)大鼠在網(wǎng)絡(luò)中行走時(shí)，計(jì)數(shù)在2分鐘內(nèi)

12、其前爪漏到網(wǎng)眼中的次數(shù)。計(jì)算公式為：(腦病變對(duì)側(cè)前爪的錯(cuò)步數(shù)-病變同側(cè)前爪的錯(cuò)步數(shù))/總步數(shù)。其分值如為正數(shù)表明腦病變對(duì)側(cè)功能缺損；如為負(fù)數(shù)表明腦病變同側(cè)功能缺損。mNSSmNSS體系的設(shè)計(jì)原理,評(píng)分越低表示功能越健全，0分表示功能完全健全的大鼠。modifiedNeurologicalSeverityScores見(jiàn)附件1轉(zhuǎn)動(dòng)桿測(cè)驗(yàn)(rotatingpoletest)轉(zhuǎn)棒上行走測(cè)評(píng):評(píng)分標(biāo)準(zhǔn)分為4個(gè)等級(jí)。0分:轉(zhuǎn)動(dòng)過(guò)程中.鼠可在棒上行走;1分:轉(zhuǎn)動(dòng)過(guò)程中.60s以上時(shí)間.鼠不會(huì)掉下來(lái);2分:轉(zhuǎn)動(dòng)開(kāi)始后.鼠從棒上掉下來(lái);3分:轉(zhuǎn)動(dòng)開(kāi)始前.鼠從棒上掉下來(lái)。10Thedetectionwasoper

13、atedat1weekaftersurgerybyinjecting0.5mg/kgofapomorphineintoabdominalcavityforobservationofthechangesinThebehavioursforonceaweekandcontinuousdetectedfor6weeks.Ifconstantright-siderotationoccurredandThenumbersofThecirclesrotatedweremorethan210rper30minutes,thoseratswereconsideredassuccessfulmodels:ifn

14、orotationoccurredorThedirectionofrotationwasnotconstantorconstantright-sideorlell-siderotationbutThespeedwasslowerthan210rper30minutes,thoseanimalswereconsideredasunsuccessfulmodels.Inadditionaltorotation,othercompanionedabnormalbehavioursalsoshouldbeobserved,suchas,tremor,sluggish,grab,andsmellexpl

15、oring,etc.rotarodtestratswereplacedonanacceleratingrotarodcylinder,andthetimetheanimalsremainedontherotarodwasmeasured.Thespeedwasslowlyincreasedfrom4to40rpmwithin5minutes.Atrialendediftheanimalfellofftherungsorgrippedthedeviceandspunaroundfor2consecutiverevolutionswithoutattemptingtowalkontherungs.

16、Theanimalsweretrained3daysbeforeMCAO.Themeanduration(inseconds)onthedevicewasrecordedwith3rotarodmeasurements1daybeforesurgery.Motortestdataarepresentedaspercentageofmeanduration(3trials)ontherotarodcomparedwiththeinternalbaselinecontrol(beforesurgery).adhesive-removalsomatosensorytestsomatosensoryd

17、eficitwasmeasuredbothbeforeandaftersurgery.Allratswerefamiliarizedwiththetestingenvironment.Intheinitialtest,2smallpiecesofadhesive-backedpaperdots(ofequalsize,113.1mm2)wereusedasbilateraltactilestimulioccupyingthedistal-radialregiononthewristofeachforelimb.Theratwasthenreturnedtoitscage.Thetimetore

18、moveeachstimulusfromforelimbswasrecordedon5trialsperday.Individualtrialswereseparatedbyatleast5minutes.Beforesurgery,theanimalsweretrainedfor3days.Oncetheratswereabletoremovethedotswithin10seconds,theyweresubjectedtoMCAO.13SpontaneousActivityTheanimalwasobservedfor5minutesinitsnormalenvironment(cage

19、).Theratsactivitywasassessedbyitsabilitytoapproachallfourwallsofthecage.Scoresindicatethefollowing:3,ratmovedaround,exploredtheenvironment,andapproachedatleastthreewallsofthecage;2,slightlyaffectedratmovedaboutinthecagebutdidnotapproachallsidesandhesitatedtomove,althoughiteventuallyreachedatleastone

20、upperrimofthecage;1,severelyaffectedratdidnotriseupatallandbarelymovedinthecage;and0,ratdidnotmoveatall.14SymmetryintheMovementofFourLimbsTheratwasheldintheairbythetailtoobservesymmetryinthemovementofthefourlimbs.Scoresindicatethefollowing:3,allfourlimbsextendedsymmetrically;2,limbsonleftsideextende

21、dlessormoreslowlythanthoseontheright;1,limbsonleftsideshowedminimalmovement;and0,forelimbonleftsidedidnotmoveatall.JulioH.Garcia,MD;SimoneWagner,MD;Kai-FengLiu,MD;Xiao-jiangHu,MD,PhD。NeurologicalDeficitandExtentofNeuronalNecrosisAttributabletoMiddleCerebralArteryOcclusioninRats。Stroke.1995;26:627-63

22、5。15ForepawOutstretchingTheratwasbroughtuptotheedgeofthetableandmadetowalkonforelimbswhilebeingheldbythetail.Symmetryintheoutstretchingofbothforelimbswasobservedwhiletheratreachedthetableandthehindlimbswerekeptintheair.Scoresindicatethefollowing:3,bothforelimbswereoutstretched,andtheratwalkedsymmetr

23、icallyonforepaws;2,leftsideoutstretchedlessthantheright,andforepawwalkingwasimpaired;1,leftforelimbmovedminimally;and0,leftforelimbdidnotmove.JulioH.Garcia,MD;SimoneWagner,MD;Kai-FengLiu,MD;Xiao-jiangHu,MD,PhD。NeurologicalDeficitandExtentofNeuronalNecrosisAttributabletoMiddleCerebralArteryOcclusioni

24、nRats。Stroke.1995;26:627-635。16ClimbingTheratwasplacedonthewallofawirecage.Normallytheratusesallfourlimbstoclimbupthewall.Whentheratwasremovedfromthewirecagebypullingitoffbythetail,thestrengthofattachmentwasnoted.Scoresindicatethefollowing:3,ratclimbedeasilyandgrippedtightlytothewire;2,leftsidewasim

25、pairedwhileclimbingordidnotgripashardastherightside;and1,ratfailedtoclimbortendedtocircleinsteadofclimbingBodyProprioceptionTheratwastouchedwithabluntstickoneachsideofthebody,andthereactiontothestimuluswasobserved.Scoresindicatethefollowing:3,ratreactedbyturningheadandwasequallystartledbythestimulus

26、onbothsides;2,ratreactedslowlytostimulusonleftside;and1,ratdidnotrespondtothestimulusplacedontheleftside.ResponsetoVibrissaeTouchAbluntstickwasbrushedagainstthevibrissaeoneachside;thestickwasmovedtowardthewhiskersfromtherearoftheanimaltoavoidenteringthevisualfields.Scoresindicatethefollowing:3,ratre

27、actedbyturningheadorwasequallystartledbythestimulusonbothsides;2,ratreactedslowlytostimulusonleftside;and1,ratdidnotrespondtostimulusontheleftside.改善記憶作用1跳臺(tái)試驗(yàn)跳臺(tái)裝置為lOcmX15cmX60cm的被動(dòng)條件反射箱，四周用黑色塑料板、底面鋪可通電的銅柵，每間內(nèi)放置一高4.5cm,直徑4.5cm的橡皮墊作為動(dòng)物回避電擊的安全區(qū)，先將動(dòng)物放在反應(yīng)箱內(nèi)的橡皮臺(tái)上適應(yīng)3min，然后立即通以40V交流電，動(dòng)物受到電擊后逃避反應(yīng)為跳上橡皮臺(tái)。記錄5mi

28、n內(nèi)大鼠受到電擊次數(shù)，稱為錯(cuò)誤次數(shù)。24h后在底部銅柵通電情況下，直接將動(dòng)物置于平臺(tái)上，記錄第1次跳下的潛伏期和5min內(nèi)的錯(cuò)誤次數(shù)。2避暗實(shí)驗(yàn)Avoidanceresponse根據(jù)大鼠有嗜暗習(xí)性分設(shè)明暗兩室。明室上方20cm處懸一個(gè)60W日光燈，在暗室的中后部分底部銅柵加40V電壓，兩室之間設(shè)有一直徑為3cm大小的圓洞。實(shí)驗(yàn)時(shí)將大鼠背對(duì)洞口放入明室，同時(shí)啟動(dòng)計(jì)時(shí)表，動(dòng)物自放入明室至進(jìn)入暗室遭電擊所需時(shí)間為潛伏期，鼠受電擊后可自行逃出暗室。記錄潛伏期及5min內(nèi)動(dòng)物受電擊次數(shù)，24h后復(fù)測(cè)。穿梭箱試驗(yàn)水迷宮試驗(yàn)（Morris）4.1Morris水迷宮為圓形水池.直徑100cm.深50cm.水深

29、30cm.水溫（24士2）C.水而覆一層塑料泡沫屑.池壁上4個(gè)等距離點(diǎn)分水池為4個(gè)象限.在第三象限中央放置平臺(tái).平臺(tái)無(wú)色透明.直徑6cm.高28cm.平臺(tái)沒(méi)于水而下2cm.水池周圍參照物保持不變。試驗(yàn)包括:定位航行試驗(yàn)（placenavigationtest）:實(shí)驗(yàn)歷時(shí)7d。將大鼠從入水面向池壁置入水槽中.記錄大鼠從入水到爬上平臺(tái)所需的時(shí)間.即逃避潛伏期。每只大鼠每人從4個(gè)不同象限入水點(diǎn)入水進(jìn)行訓(xùn)練各1次取其平均值為其每人逃避潛伏期。訓(xùn)練中若大鼠在60s內(nèi)找到平臺(tái)讓其于平臺(tái)上站立lOs;若未找到用棒將其引上平臺(tái)并讓其站立10s.潛伏期限記為60s。將大鼠從平臺(tái)上拿卜來(lái)休息60s之后.再按序由下

30、一入水點(diǎn)入水進(jìn)行下一次實(shí)驗(yàn)?？臻g探索試驗(yàn)(spatialprobetest)第8天撤去平臺(tái).將大鼠從第二象限入水點(diǎn)放入水槽.記錄60s內(nèi)其在平臺(tái)象限的滯留時(shí)間。4.2水迷宮試驗(yàn)主要用于測(cè)試大鼠對(duì)空間位置覺(jué)和方位覺(jué)的學(xué)習(xí)記憶能力5水迷宮試驗(yàn)測(cè)試的一個(gè)重要指標(biāo)是大鼠的平均逃避潛伏期平均逃避潛伏期指的是大鼠在放入迷宮后學(xué)會(huì)在5中內(nèi)找到平臺(tái)所需要的平均時(shí)間,主要與海馬功能有關(guān),與大腦皮層也有一定的關(guān)系。MorrisWaterMazeThisisataskthatmeasureslearningandmemoryinanaqueousenvironment.Briefly,theratwasplaced

31、ina4-ft-diameterwatertankthatisvisuallyseparatedintofourquadrants.Inthecenterofonequadrant,thegoalquadrant,aplatformishidden1inchbelowthewaterline.Theratwasplacedinthetankatoneoffourstartlocations(north,south,east,west)andswamforupto60secondsuntilitfoundtheplatform.Oncetheanimallocatedtheplatform,it

32、remainedonitfor20seconds.Intheeventthatananimaldidnotlocatetheplatform,theobserverpositionedtheanimalontheplatformattheendofthe60-secondswimperiod.Duringa2-to4-minuteintertrialinterval,theanimalswerekeptwarmbyinfraredheatinglamps.Thetemperatureofthewatermazewas200.5C.Thelatencyandpathlengthoftheratw

33、eremeasuredbyavideotrackingsysteminterfacedtoacomputer.NancyE.Alexis,MS;W.DaltonDietrich,PhD;EdwardJ.Green,PhD;RicardoPrado,MD;BrantD.Watson,PhD.NonocclusiveCommonCarotidArteryThrombosisintheRatResultsinReversibleSensorimotorandCognitiveBehavioralDeficits.Stroke.1995;26:2338-2346.4.3Y迷宮學(xué)習(xí)記憶訓(xùn)練及測(cè)試:Y型電

34、迷宮臂長(zhǎng)30cm.高15cm.寬15cm。每臂末端有15W信號(hào)燈示安全區(qū).箱底銅柵間隔4mm。電流強(qiáng)度0.7mA.選用電壓38V.電擊延時(shí)2s。設(shè)一臂為起步區(qū)按I-II-III-I臂順序輪流作為安全區(qū)。大鼠在起步區(qū)靜置3min.予以電擊致其逃至安全區(qū)燈光持續(xù)15s.然后熄燈體急45s.開(kāi)始下一次操作。學(xué)習(xí)訓(xùn)練及測(cè)試:規(guī)定大鼠受電擊后從起步區(qū)直接逃至安全區(qū)為“正確反應(yīng)”.以達(dá)到連續(xù)10次電擊均為正確反應(yīng)(即10/10)前所需的電擊次數(shù)(即嘗試次數(shù))表示其學(xué)習(xí)獲得能力(acquisition)o若嘗試次數(shù)超過(guò)30次則不再測(cè)試.并以30次為最大值計(jì)數(shù)。記憶再現(xiàn)(retrieval)測(cè)試:選上述達(dá)到1

35、0/10標(biāo)準(zhǔn)的大鼠.休息24h后再次同法測(cè)試.以達(dá)10/10標(biāo)準(zhǔn)前的嘗試次數(shù)表示記憶再現(xiàn)能力。Motorbehaviorobservationofspontaneousipsilateralcircling,gradedfrom0(nocircling)to3(continuouscircling);contralateralhindlimbretraction,whichmeasuredtheabilityoftheanimaltoreplacethehindlimbafteritwasdisplacedlaterallyby2to3cm,gradedfrom0(immediaterepla

36、cement)to3(replacementafterminutesornoreplacement);beamwalkingability,graded0foraratthatreadilytraversesa2.4-cm-wide,80-cm-longbeamto3foraratunabletostayonthebeamfor10seconds;平衡木測(cè)驗(yàn)(balancebeamtest)平衡木長(zhǎng)80cm，寬2.5cm的方木棒，平放在距離地面l0cm處，讓大鼠在其上行走。評(píng)分標(biāo)準(zhǔn)為6個(gè)等級(jí)。0分:能跳上平衡木，在上面行走不會(huì)跌倒;T分:能跳上平衡木，在上面行走跌倒機(jī)會(huì)小于50；2分：能跳上平衡木，在上面行走跌倒機(jī)會(huì)大于50；3分：在健側(cè)后肢幫助下能跳上平衡木，但受累癱瘓側(cè)后肢不能幫助向前移動(dòng)；4分：在平衡木上不能行走，但可坐在上面；5分：將大鼠放在平衡木上會(huì)掉下來(lái)。Beambalanceperformanceofanimalswasassessedona6-pointscale17:0=balanceswithsteadyposture;1=graspssideofbeam;2=hugsbeamand1limb,fallsdownfrombeam;3=hugsbeamand2limbs,fallsd