Delayed administration of TAT.ARC protein attenuates focal ischemic brain injury and fosters recovery in the long-term: behavioral data of neuroscores and rota rod tests, daily temperature and body weight measures

Mice were subjected to 30 min MCAo or sham surgery and TAT.b-Gal or TAT.ARC were administered in the contralateral ventricle after 3h after the onset of ischemia and mice were observed over 28 days. DeSimoni Neuroscore was performed at indicated time points (Fig. 8) as described (De Simoni et al., 2003; Orsini et al., 2012) with some modifications. In brief, general health (Table 2) and specific focal assessments (Table 3) were separately scored, analyzed and finally added to form a summation score. Summative scores added up to a maximum of 43 points with more points meaning more deficits. Rota rod was assessed as described recently (Hoffmann et al., 2015) and the best run out of three replicates at a given time point was used for statistical analysis. Body temperature was measured daily as survival was documented. Body temperature was non-invasively assessed at the same time of the day prior to body weight measurements using subcutaneous transponders (IPTT-300, Bio Medic Data Systems), for unambiguous identification of mice in their home cages as described (Kort et al., 1998; Langer and Fietz, 2014).

将小鼠予以30分钟大脑中动脉闭塞（Middle Cerebral Artery Occlusion, MCAO）造模或假手术，并在脑缺血发作后3小时，于对侧脑室内给予TAT.b-Gal或TAT.ARC，随后对小鼠进行为期28天的观察。DeSimoni神经功能评分（DeSimoni Neuroscore）于指定时间点开展测定（如图8所示），参照De Simoni等（2003）、Orsini等（2012）的经典方案并做部分修改。简言之，将一般健康状况（表2）与特异性局灶性评估（表3）分别进行评分、分析后求和，得到总评分。总评分最高为43分，分值越高提示神经功能缺损程度越重。转棒实验（rotarod test）参照Hoffmann等（2015）的最新方案进行评估，取指定时间点下三次重复测试中的最优结果用于统计分析。每日记录小鼠体温与存活状态。体温采用无创方式检测：于每日固定时间、体重测量前，使用皮下植入式应答器（IPTT-300, Bio Medic Data Systems）进行测定，该方法可在小鼠饲养笼中实现个体的精准识别，参照Kort等（1998）、Langer与Fietz（2014）的研究方法。