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).

将小鼠接受30分钟大脑中动脉阻塞（Middle Cerebral Artery Occlusion, MCAO）造模或假手术，于缺血发作后3小时向对侧脑室注射TAT.β-半乳糖苷酶（TAT.β-Gal）或TAT.ARC，随后连续观察28天。DeSimoni神经评分（DeSimoni Neuroscore）按照已发表方法（De Simoni等，2003；Orsini等，2012）稍作修改后，于指定时间点开展评估（图8）。简言之，实验分别对小鼠一般健康状态（表2）与特异性局灶性神经功能评估项（表3）进行评分、分析，最终求和得到总评分；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（Rota rod）参考近期研究方法（Hoffmann等，2015）进行，取同一时间点三次重复测试中的最优成绩用于统计学分析。每日测量小鼠体温并记录生存情况，体温采用非侵入式方式检测：于每日固定时间、体重测量前，使用皮下应答器（IPTT-300, Bio Medic Data Systems）进行检测，该方法可精准识别饲养笼内的小鼠，具体操作参见已发表文献（Kort等，1998；Langer和Fietz，2014）。