各种神经行为学评价

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(behavior test)的十分重要，对其评定的方法也颇多，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3位参加试验的人员分别以单盲法对试验的大鼠进行打分和记录.然后将3组的记分结果进行平均后的得分进行统计计算。

（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)1. 神经行为学检查Longa评分法2．Berderson评分法姿势反射测验（postural reflex test）3. 攀绳实验4．网屏测验（screen test）5．肢体放置测验（limb-placement test）Elicited Forelimb Placing6．开野试验（Open－Field法）测定行为7．MNSS8.转动杆测验（rotating pole test）9．Rotation10．肢体对称试验评分法11. rotarod test12. adhesive-removal somatosensory test13．Spontaneous Activity14．Symmetry in the Movement of Four Limbs15．Forepaw Outstretching16．Climbing17．Body Proprioception18. Response to Vibrissae Touch改善记忆作用1.跳台试验2.避暗试验3.穿梭箱试验4.水迷宫试验Motor behavior(1) observation of spontaneous ipsilateral circling,(2) contralateral hindlimb retraction,(3) beam walking ability,平衡木测验（balance beam test）(4) bilateral forepaw grasp,Skilled forelimb function(1)staircase feeding apparatusnociception(1) plantar test1．神经行为学检查:Longa评分法神经学检查分5个等级：0分：正常，无神经功能缺损；1分：左侧前爪不能完全伸展，轻度神经功能缺损；2分：行走时，大鼠向左侧（瘫痪侧）转圈，中度神经功能缺损；3分：行走时，大鼠身体向左侧（瘫痪侧）倾倒。

重度神经功能缺损；4分：不能自发行走，有意识丧失。

2．Berderson评分法姿势反射测验（postural reflex test）提尾悬空试验大鼠提尾悬空一米，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0分。

梗死后大鼠持续屈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1分。

大鼠被放于大的软垫子上，表面有大鼠可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2分。

然后让大鼠自由活动，观察转圈行为。

大鼠持续向瘫痪侧转圈得3分。

前肢屈曲带有侧推抵抗力下降经常见到；双上肢屈曲并侧推抵抗力下降大鼠经常有转圈行为。

神经检查在3－5分钟内完成。

正常 0分未见行为缺陷中度 1分前肢屈曲(即提尾悬空实验阳性)重度 2分侧推抵抗力下降(即侧向推力实验阳性).伴前肢屈曲.无转圈行为3分同2级行为.伴自发性旋转。

提起鼠尾.使其离地0. 33 m高.观察前肢情况。

正常大鼠两前肢对称向前伸开;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4分.正常评分为0分。

将动物置于平滑地板上.分别推其左、右肩向对侧移动，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1－3分.正常评分为0分。

观察两前肢肌力。

将动物两前肢置于一金属网上.然后轻提起大鼠.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1－3分.正常评分为0分。

根据上述行为检查的情况.积分值为0一10分。

积分越高说明动物行为障碍越严重。

此测验对皮层和纹状体的损伤较敏感。

3攀绳实验: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1. 5 cm的麻绳从高1 m、长和宽分别为15、50 cm的平台上垂下，手术前1d训练每只大鼠.使之能够爬到绳子的上1/ 3。

每个测试日测试3次。

评分的标准如下:0分.不需任何刺激在10s内爬上平台;1分.不需任何刺激在15 s内爬上平台;2分.刺激次数小于5次.在30 s内爬上平台;3分.刺激次数小于5次.在60 s内爬上平台;4分.刺激次数大于5次.在60 s内爬上平台或是刺激小于5次.但爬上平台时间超过60 s; 5分.大鼠不能爬上平台。

4．网屏测验（screen test）网屏训练:网屏为50cm X 40cm网带.网眼为lcm X lcm.网板的左右和上方都用25cm高的木板框边.网屏距地面高度为80cm.下方铺以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2s内将此屏风变成垂直位.保持5s.观察大鼠是否会从网屏上下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4个等级。

0分:前爪握住网屏达5s之久，不会掉下来;1分:暂时握住网屏，滑落一段距离，但没有掉下来;2分:在5s内掉下来;3分:网屏转动时，鼠即刻掉下来。

5．肢体放置测验（limb-placement test）Elicited Forelimb Placing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Ｋａｗａｍａｔａ等[2]采用的动物行为测试评分方法。

具体实验有: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10ｃｍ处向桌面缓慢斜线靠近(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。

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。

(2)触觉亚实验,将动物置于桶状凹陷实验器内,使其前爪悬空,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。

(3)本体觉亚实验,操作及评分同触觉亚实验,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0～10分,功能损伤越重,得分越高。

6．开野试验（Open－Field法）测定行为方法：本实验所用敞箱为正方形,长、宽各80cm,高40cm,底面由面积相等边长16cm的正方形25块组成,以动物穿越底面的块数为水平活动(crossing)得分,以直立次数为垂直活动(rearing)得分,每只动物共进行7次测定,每次3minOpen2Field法是测定抑郁症大鼠行为学的经典方法,分水平运动和垂直运动2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动评分及垂直运动评分。

Elevated open field activity is characteristic of hippocampally damaged animals19 20 21 22 and has been observed in animals with striatal/cortical damage as well.23 Rats are placed in a novel environment (3x3 ft) with 3-inch-square grid markings on the base. The number of times a rat crosses a grid line is recorded for a 5-minute period.7．肢体对称试验评分法将大鼠置于网中，网眼大小为2.3 x2.3cm,当大鼠在网络中行走时，计数在2分钟内其前爪漏到网眼中的次数。

计算公式为：（脑病变对侧前爪的错步数－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．mNSSmNSS体系的设计原理,评分越低表示功能越健全，0分表示功能完全健全的大鼠。

modified Neurological Severity Scores见附件19.转动杆测验（rotating pole test）转棒上行走测评:评分标准分为4个等级。

0分:转动过程中.鼠可在棒上行走;1分:转动过程中.60s以上时间.鼠不会掉下来;2分:转动开始后.鼠从棒上掉下来;3分:转动开始前.鼠从棒上掉下来。

10．The detection was operated at 1 week after surgery by injecting 0. 5 mg/kg of apomorphine into abdominal cavity for observation of the changes in The behaviours for once a week and continuous detected for 6 weeks. If constant right-side rotation occurred and The numbers of The circles rotated were more than 210 r per 30 minutes, those rats were considered as successful models: if no rotation occurred or The direction of rotation was not constant or constant right-side or lell-side rotation but The speed was slower than 210 r per 30 minutes, those animals were considered as unsuccessful models. In additional to rotation, other companioned abnormal behaviours also should be observed, such as, tremor, sluggish, grab, and smell exploring, etc. 11. rotarod testrats were placed on an accelerating rotarod cylinder, and the time the animals remained on the rotarod was measured. The speed was slowly increased from 4 to 40 rpm within 5 minutes. A trial ended if the animal fell off the rungs or gripped the device and spun around for 2 consecutive revolutions without attempting to walk on the rungs. The animals were trained 3 days before MCAO. The mean duration (in seconds) on the device was recorded with 3 rotarod measurements 1 day before surgery. Motor test data are presented as percentage of mean duration (3 trials) on the rotarod compared with the internal baseline control (before surgery).12. adhesive-removal somatosensory testsomatosensory deficit was measured both before and after surgery. All rats were familiarized with the testing environment. In the initial test, 2 small pieces of adhesive-backed paper dots (of equal size, 113.1 mm2) were used as bilateral tactile stimuli occupying the distal-radial region on the wrist of each forelimb. The rat was then returned to its cage. The time to remove each stimulus from forelimbs was recorded on 5 trials per day. Individual trials were separated by at least 5 minutes. Before surgery, the animals were trained for 3 days. Once the rats were able to remove the dots within 10 seconds, they were subjected to MCAO.13．Spontaneous ActivityThe animal was observed for 5 minutes in its normal environment (cage). The rat's activity was assessed by its ability to approach all four walls of the cage. Scores indicate the following: 3, rat moved around, explored the environment, and approached at least three walls of the cage; 2, slightly affected rat moved about in the cage but did not approach all sides and hesitated to move, although it eventually reached at least one upper rim of the cage; 1, severely affected rat did not rise up at all and barely moved in the cage; and 0, rat did not move at all.14．Symmetry in the Movement of Four LimbsThe rat was held in the air by the tail to observe symmetry in the movement of the four limbs. Scores indicate the following: 3, all four limbs extended symmetrically; 2, limbs on left side extended less or more slowly than those on the right; 1, limbs on left side showed minimal movement; and 0, forelimb on left side did not move at all.Julio H. Garcia, MD; Simone Wagner, MD; Kai-Feng Liu, MD; Xiao-jiang Hu, MD, PhD。