We would like to thank everyone at Shriners Hospital Pediatric Research Center, in particular Dr. Mickey Seltzer, of whom without his support, this work would not have been funded. In addition, we would like to thank Isha Srivastava, who contributed to early data collection and Amy He, who helped with the figures. This study was funded by Shriners Hospitals for Children. No funding source played a role in experimental design or decision to submit the paper for publication.

注:所有动物手术均按照坦普尔大学的ULAR部门和IACUC政策和程序进行。 C57BL / 6水坝和公牛从Charles River实验室购买并饲养在笼子里饲养了12小时的光/暗周期（灯亮7:00 - 晚上7:00），免费获得食物和水。对繁殖产生5之间产羔 - 10幼崽。 1.脑瘫手术诱导<ol><li>注:脑瘫使用产后天（PND）6小鼠的幼崽引起的，如前面所述8,9（http://jove.unicartagenaproxy.elogim.com/video/1951/mouse-models-of-periventricular-leukomalacia）。 </li><li>放置在小狗在冰玻璃碗里用实验室擦拭，以保护小狗的皮肤。步行捏和缺乏运动检查适当的麻醉平面。移动小狗到填充冰袋进行手术。 </li><li>消毒用70％的乙醇的小狗的皮肤上。干燥时，可使用11＃无菌手术BLA德和使在颈部1厘米的切口。 </li><li>使用立体手术显微镜，隔离用小钩的右颈总动脉，并用便携式手持cauterizer烧灼。目视确认该动脉闭塞。假手术包括可视化和颈总动脉的隔离而不烧灼。 </li><li>重新调整皮肤和使用紧密缝合胶（正氰基丙烯酸正丁酯）。 </li><li>放置在34℃下加热垫的小狗30分钟监视自主呼吸和正常运行。 </li><li>返回小狗（S）大坝30分钟。 </li><li>放置幼仔在加热垫或其他加热装置低氧室35分钟设定在6％的氧气内部设定在34℃下。氧被氮置换。密切监察一致的伤害结果室内氧气含量和温度。 </li><li>从缺氧室中取出幼崽，并将其返回加热垫。 </li><li>腹腔注射脂多糖糖在无菌盐水中以1微克/千克稀释，小狗返回大坝。深水注射是唯一的盐水注射。 </li></ol> 2.新生儿电机测试注:在PND 8，48小时CP诱导后，小鼠的幼崽是为神经行为发育测试。幼仔一个4小时块内中午前，以消除行为的差异一天时间的考验。幼仔从坝为不超过15分钟，在一个时间，以防止体热和饥饿/分离问题迅速丧失除去。此外，幼崽被允许测试之间的休息，这样最大的努力将在每个测试被引出。新生儿马达试验的基础是一种适于使用的测试10,11-福克斯的电池和福克斯的测试12 Wahlsten的适应性，以及款待-NMD和其他行为的出版物（如文中对于每个测试说明）。测试福克斯的电池适用于PND 2 - 21福克斯的测试，面糊Ÿ在座的包括:翻正反射，抓握反射，负趋地（称为狐狸的电池竖屏测试）和四个肢体握力（从福克斯和Wahlsten的屏幕爬坡测试修改）。在这里，步行，前下肢肌力，及后肢力量也被测试来区分虚假和CP的乳鼠之间的反身性的运动行为。为了消除在测试由于学习改进，试验仅限于指出最多3试验。所有其他测试过每个动物只有一个审判。 <ol><li> 行走 （ 图1）（改编自大鼠协议13）: 注:抓取是在PND 0之间的鼠标小狗早期开发的行为- 5，在该点的小鼠开始过渡到步行，从5 - - 10日龄14。在PND 8，下地测试需要这个过渡时间过程的优势。下地可以，但是，在整个鼠标的寿命得分，可以在任何年龄来确定。至于存在用于学习没有潜在的行走试验可作为通过实验的过程中根据需要重复多次。 <ol><li>在透明外壳地方小鼠，其中小鼠是可见从顶部以及侧。通过触摸小狗的尾巴用轻微的针刺激励小狗走路。 </li><li>分数下地使用下列标准3分:0 =无动作，1 =非对称肢体运动爬行，2 =缓慢爬行，但对称的肢体运动，而3 =快速爬行/行走。 注意:在此，对称肢体运动被描述，其中后爪在每个步骤中满足frontpaws，并且每个步骤顺利过渡到下一步骤。鼠标显示不对称肢体运动具有不稳定的爪子安置和过渡从一步到下的并不顺利。 </li></ol></li></ol><img alt="图1" src="/files/ftp_upload/53569/53569fig1.jpg" /> 图1.从转变爬行到散步可以通过观察后爪，以及在头部和尾部进行区分:（A）在爬行，整个背部爪，从脚趾到脚跟，走动时接触地面，并标示为（*）。一个成年人的步行模式观察时只的脚趾和前部的后爪接触地面（脚跟升高时，由[**] deonoted）。（B）中的头部和一爬行小鼠的尾部低到地面。头开始从爬行到行走在过渡期间上升。过渡完成后，无论是头部和尾部升高，只有后肢的前部接触地面。 <a href="https://jove.unicartagenaproxy.elogim.com/files/ftp_upload/53569/53569fig1large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> <ol start="2"><li> 后肢足角 （ 图2） 注:在后肢姿势作为谅解备忘录的明显变化发展E从爬行到行走，其中该后肢被身下行走和后肢之间的角度小于在爬行看到的角度定位时的成熟。即使后肢足角随时间变化，同样的年龄不同伤害或疾病的小鼠的幼崽可以比拟的。类似于步行测试（3.1），对于学习的可能。因此，如通过实验的过程中所需要的后肢脚角度测试可以重复多次。 <ol><li>无论是在明确公开字段框或封闭区域，从下方或上方，分别安装摄像机，记录小狗，因为它绕着字段。通过触摸小狗的尾巴用轻微的针刺激励小狗走路。记录两分钟。 </li><li>使用录像，通过从脚后跟/胫最长（中）脚尖的端画线测量幼崽的脚角度。只有当小狗正在执行一个完整的步幅进行测量在一条直线上并且两个脚平放在地面上。而小狗是固定的，不要进行测量或在小狗正在转向。 </li><li>测量三个五套脚的角度，计算所测试的各小狗平均角度。 </li></ol></li></ol><img alt="图2" src="/files/ftp_upload/53569/53569fig2.jpg" /> 图2.后肢足角度可用于确定步态异常，脚的角度可以通过中间的（最长）数字绘画从脚跟中间线来衡量。相比普通（见代表性的成果，脚角度）时，受伤的动物有更大的脚的角度。 <a href="https://jove.unicartagenaproxy.elogim.com/files/ftp_upload/53569/53569fig2large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> <ol start="3"><li> 面翻正 （ 图3）: 注:翻正反射是运动能力<html>鼠标小狗，以便能够从仰卧位置翻转到它的脚。为正位反射出现在啮齿动物的平均年龄是PND 5从PND 1的范围内- 10 15本试验是一种反射，没有学习组件，它可以在整个实验期间进行重复。 <ol><li>放置在棉片或板凳垫在背上的幼崽，并保持在适当位置，持续5秒。 </li><li>释放幼仔并记录所花费的小狗返回到俯卧位置的时间，以及扶正的方向（左或右）。共1分钟，给出了每个试验中，如果需要的话。 </li><li>重复一共有三个试验。 </li></ol></li></ol><img alt="图3" src="/files/ftp_upload/53569/53569fig3.jpg" /> 图3.表面扶正。这个测试需要躯干控制，并可能测试姿势不平衡。人类CP患者可能有自己的核心赤字。e.com/files/ftp_upload/53569/53569fig3large.jpg"目标="_空白"&gt;点击此处查看该图的放大版本。 <ol start="4"><li> 负趋地性 （ 图4） 注:平均年龄为负趋地反射出现在啮齿动物是PND 7与PND 3的范围- 15 15负趋地试验评估了年轻小鼠运动协调。小鼠置于朝下的斜面，并且由于重力的前庭线索，幼仔转向面对的斜坡。应对刺激，或出租车，是一种与生俱来的行为。 <ol><li>将小狗与它的头在45 度斜面朝下，并保持5秒。 </li><li>松开小狗并记录小狗转向朝上的时间和方向。总测试时间为2分钟。 </li><li>重复一共有三个试验。那倒下的倾斜或无法开机的小鼠可以是重新测试，淘汰，或给予零分。 注:这一决定为lEFT考官，因为偶尔会幼仔斜面滚下因嗜睡而不是弱点。一旦决定如何得分的倒下倾斜幼仔提出，应该在方法要注意，应该在所有科目的测试是一致的。 </li></ol></li></ol><img alt="图4" src="/files/ftp_upload/53569/53569fig4.jpg" /> 图4.负趋地，电机和前庭输入所需的鼠标来识别一个斜坡的方向和转身。 <a href="https://jove.unicartagenaproxy.elogim.com/files/ftp_upload/53569/53569fig4large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> <ol start="5"><li> 前悬挂肢 （ 图5）16;改编自17，18: 注:前悬挂肢体幼仔测试，包括手臂和爪强度的前肢力量。 ŧ不推荐他的测试比幼仔PND 10 15岁。幼仔把握不准跨越一个稳定的对象串成一个电线挂到与两个前爪电线。测试面积超过软垫放置区。该测试可以检测左/右侧的实力差异。学习和没有负筋可导致增加的非参与。小鼠释放或当放置在电线未能把握时立即降指示不参与。 <ol><li>人体握紧幼仔，使他们与两个前爪抓住电线。 </li><li>松开小狗。使用定时器或秒表，记录的总时间下跌，以及爪子的弱点。 注意:如果小狗始终从一只爪子电线落在之前，其他而不是在同一时间两个爪子拉线释放爪子弱点是确定的。 </li><li>重复试验，共三次。 </li></ol></li></ol> 图5.前肢悬挂。这悬浮试验引起前肢紧张，直到肌肉疲劳。通过这种方法，在前肢基线强度成立。 <a href="https://jove.unicartagenaproxy.elogim.com/files/ftp_upload/53569/53569fig5large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> <ol start="6"><li> 后肢悬架 （ 图6）: 注:本悬架测试确定后肢力量。它是专为新生儿设计了一个试验，最初PND 2之间使用动物- 12 19,20，但可以适用于小鼠高达PND 14这个测试可检测左/右后肢强度差异以及神经肌肉功能。标准的50毫升锥形被使用，实验室抹布填充。类似于前肢悬浮试验，该试验中可以得知，尤其罪CE没有负面后果下降。因此，增加了非参与，一旦落小鼠为视为释放或不留时放置在管的边缘，可以注意到。 <ol><li>用50毫升的锥形，地点小狗轻轻面朝下放入管后腿挂在篮筐。 </li><li>松开小狗。观察后肢的姿势。 </li><li>按照下列标准得分的姿势。 注:4分表示尾部提出后肢正常分离; 3得分手段的弱点是明显，后肢更接近在一起，但他们很少互相接触; 2分表示后肢接近，彼此经常接触; 1比分显示的弱点是明显，后肢几乎总是与提出的尾部紧握位置; 0分表示与降低尾部或未能守住管的任何一段时间的后肢不断扣紧。 </li><li>如有必要，伯爵拉。一拉就是资格赛当指明分数小狗尝试使用它的后肢提起它的身体，而在锥形管的一侧暂停。 </li><li>使用定时器或秒表，记录潜伏期下降。 </li><li>重复一式三份整个测试。 </li></ol></li></ol><img alt="图6" src="/files/ftp_upload/53569/53569fig6.jpg" /> 图6.后肢暂停。（A）。此悬浮试验导致后肢紧张，直至肌肉疲劳。在后肢基线强度和姿势的建立。（B）。计分。注意代表的小鼠表现出可能的姿势评分上面的数字。 <a href="https://jove.unicartagenaproxy.elogim.com/files/ftp_upload/53569/53569fig6large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> <ol start="7"><li> 握力 （ 图7）: 注意:此测试将检查爪子小号同时四只爪子的trength。一个16×18的屏幕玻璃纤维丝被使用。对啮齿动物能够掌握一个横屏的平均年龄是PND 8的范围内的PND 5 - 15 15 福克斯使用的四种肢横屏试验从PND 2 - 21 10本试验是从标准的水平改性屏幕测试;这里的画面从水平到垂直位置，缓慢转动，以挑战所有四肢21的把持;改编自尔蒂第16如果当反转180°鼠标保持到网筛，记录延迟下降。另外，还要注意体重。一辆挂冲动可以计算为[重量（g）×潜伏期下降（SEC）反映抗拒地心引力所需要的力量。 <ol><li>使用一块金属丝网，将小狗在屏幕上。让小狗适应这样的环境，时间约为5秒。 </li><li>缓慢倒置屏幕180度。记录的大致ANGL屏幕的E当小狗脱落。 </li><li>重复一共有三个试验和平均的审判。 </li></ol></li></ol><img alt="图7" src="/files/ftp_upload/53569/53569fig7.jpg" /> 图7.握力。小鼠需要维持四肢的引力增大肌肉紧张。 <a href="https://jove.unicartagenaproxy.elogim.com/files/ftp_upload/53569/53569fig7large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> <ol start="8"><li> 把持反射 （ 图8） 注:抓握反射通常在PND 7显示在啮齿动物中与来自PND 3的范围- 15 15每个爪被单独测试，从而测试可以揭示前置式或后肢的问题，以及片面性的问题。由于它是一种反射，这种测试可以重复，直到出现反射。这是不容易学习。作为一个重要的警告，该测试<html>不区分抓实力，只有能力，而且必须在幼年小鼠开始掌握因害怕应对15日龄前进行测试。 <ol><li>它的脖子颈背，类似于鼠标小狗被大坝携带的方式握住鼠标。这使得持有小狗成为本能地不动，宽松，允许便于测试。 </li><li>中风与直率的小狗爪子每一个，圆刀片的一侧。 </li><li>单独测试每个爪子抓记录的存在或不存在和得分每个爪子1点与鼠标抓手。 注:右爪偏好的评分是右爪偏爱100％， - - -100％左爪子偏好，两个爪子50％的把握，而对于没有爪子抓0％。以确定这些数字的公式为[（右爪 - 左爪）/（右爪+左爪子+两者爪子）]×100％。 </li></ol></li></ol>/53569/53569fig8.jpg"/&gt; 图8.抓握反射。由于新生小鼠没有强烈的恐惧反应，本次测试严格确定跖/手掌反射。 <a href="https://jove.unicartagenaproxy.elogim.com/files/ftp_upload/53569/53569fig8large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> <ol start="9"><li> 崖厌恶 （ 图9）: 注:崖厌恶测试迷宫反射，以及强度和协调，并可以用来从PND 1-14 22测试幼仔。预香味中（其中至少5只小鼠的已被允许一个框来自由漫游）用平升高的壁架的使用量和小狗被置于与数字只前爪和定位在边缘的口鼻部。得分是通过计算花费的小狗把从悬崖路程，移动它的爪子和鼻子远离边缘的总时间进行。如果没有响应30秒钟后看到的那样，试验被终止。如果小狗脱落的边缘，可以执行单个附加试验。 <ol><li>使用的侧视图，放置在预香味框的边缘的小狗，确保前脚掌，数字和吻是在边缘的唯一零件。 </li><li>放开小狗，并开始计时。 </li><li>一旦两个鼻子和爪子已经从边缘去除，停止计时和记录时间。 </li><li>重复试验，共进行3次。如果小狗没有在30秒内摆脱悬崖而去，没有得分给出。 注:小狗是一个非参与者对受损的是留给考官的自由裁量权的决心。悬崖的高度可以为年龄小狗，以保证小狗的安全性进行调整。一个较小的高度可以与黑"地板"用于模拟一个更高的高度。 </li></ol></li></ol><img alt="图9" src="/files/ftp_upload/53569/53569fig9.jpg" /> FIGURË9.悬崖厌恶。前庭失衡正在使用的悬崖厌恶测试测量。在这里，小狗的眼睛仍然关闭，怕是没把从悬崖边走的驱动因素。 <a href="https://jove.unicartagenaproxy.elogim.com/files/ftp_upload/53569/53569fig9large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> 3.统计学意义<ol><li>使用统计软件分析结果。表示该数据为平均值（SEM）的平均值±标准误差。测试参数，因此，检查用t检验分析数据。 注:实验的目的不是测试性别差异。差异被认为是当p &lt;0.05统计学显著。 </li></ol>

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The authors declare that they have no competing financial interests.

使用动物模型来研究人类疾病是否存在人类和啮齿动物之间，并且所执行的行为测试有直接关系到人类的症状的细胞和分子响应之间的重叠是唯一相关。之一的儿科疾病的研究的主要问题是，许多研究者使用成年啮齿动物创建模型，以及成年啮齿动物行为的评价，在不考虑发育差异可能对疾病过程重要。由于这些问题，重要的是在儿科疾病使用研究不仅适当调整的发育时间点（ 例如，人的中枢神经系统发育，在28 - 51周内，相当于一个产后第2天-第7天的啮齿动物）7，而且行为测试，将审查适当的运动，感觉或反射性发育行为。因此，因为每个新的新生儿疾病模型开发，它必须经过严格的测试，以确保蜂窝和行为反应将提供啮齿类动物和人类之间最合适的翻译资料。 脑瘫是一种运动障碍，它持续到成年。许多当今脑瘫模型的一个问题是缺乏可重复的，标准化的电机测试，可以在儿科患者看到的赤字相关的。在这种新的模式，它结合缺氧，缺血和炎症的新生小鼠，运动行为，用具体的新生小鼠试验电池进行评估。为了减小主体和增加定量报告，几个测试已被修改，以包括非常具体的，但容易评估，可以标准化的措施。此外，前端和后肢评价可以单独执行，并且左/右差异可被确定。测试该电池是特异于新生小鼠最多两个周龄。 这种CP模型演示行走困难（行走，后肢足角），以及肢体特异性无力（4肢悬浮，后肢悬挂），和赤字发育反射（抓握反射）。尽管在本研究中检测只有一个时间点，这些缺陷可以随时间被跟踪。 有测试其他电池可以在新生儿中使用，如测试的福克斯的电池或发育里程碑15的Heyser的评价。然而，这些测试的新生儿进行比较的成人，其响应可能是不一样的，因为新生儿仍在发展。福克斯的电池和Heyser的Assement测试依赖于观测主观信息用二分（是或否）的评估，而不是客观的数据（角度，姿势的基础上强度等 ）。由于这些测试的主观性，许多科学家已经适应，添加或删除的标准，从而使他们的结果无比给他人，并限制ING在对于特定疾病或病症建立基线赤字方面的数据的有用性。通过建立一组标准化电机测试是定性和专为测试新生儿，从单个研究小组结果能够精确和可靠地报告和比较。

开发儿童受伤或利用啮齿类动物疾病的新车型往往是困难的，因为大鼠和小鼠的惊人的能力，迅速从神经损伤恢复。因此，为了验证任何新的儿科疾病模型，深入研究细胞和分子的变化必须去手牵手与行为的结果。在许多方面，功能行为恢复可能比在治疗或翻译相关性方面潜在的细胞的变化更为重要。随着研究人员更多地了解在成人和新生儿损伤，很显然，他们的反应是非常不同的，两者不能外推。例如，新生小鼠显示不同水平的神经生长因子，脑源性神经营养因子，神经营养因子-3和脊髓损伤后1,2胶质细胞源性神经营养因子。此外，有新生儿中风后3，D显著血脑屏障泄漏emonstrate皮层神经元重排周围神经损伤后4，并有脊髓损伤和缺血缺氧5,6延迟或减缓星形胶质细胞增生。因此，平移儿科研究使用发育等效模型和这些模型是为两个蜂窝/分子变化和年龄适当行为测试评价是很重要的。 脑瘫（CP）是一种运动障碍，影响3:1000名活产儿每年（NIH）。带CP的儿童显示出一系列的症状和共病的条件下，根据疾病的严重程度。难度与运动和协调是最常见的迹象，在达到马达发育标志延迟沿。其他症状包括肌张力异常（增加或减少），降低的精细动作技能，行走困难，流口水过多和吞咽，和语音延迟（NIH）。 CP的根本原因被认为是在芽前或围产期期间，或最多一年产后缺乏氧和/或血液流向脑部的。此外，炎症现在认为是在CP的发展的重要组成部分。 大多数CP案件与周围的脑室，被称为脑室周围白质软化症（PVL）脑白质损伤有关。这种神经标志表明，最初的侮辱导致CP期间大脑发育的时期，少突胶质细胞最容易受到侮辱发生。快速少突胶质细胞的生长在一个人的期间，也期间时少突胶质细胞是最容易受到伤害，是24之间 - 周32妊娠。在啮齿动物，等效周期是产后天7月2日至7日，并且是当CP在这个模型中被诱导。 这是用于进行此处所述的测试CP的新生小鼠模型结合缺氧和局部缺血与炎症创建一个注射URY，更好地模仿神经退行性疾病在人类CP看到。该模型解决一些在CP的其他动物模型中观察到的大的缺点，缺乏鲜明运动缺陷类似于人类CP患者，以及不同的白质损伤。通过使用相同的模型协作者以前的研究已经表明，在加入炎症增强脑白质损伤，从而更好地模仿在儿童看到具有CP 8的PVL。在以前的数据的基础上，提出了以评估在运动行为变化动物年龄的新生儿电机测试的综合电池。

<table><tbody><tr><td></td><td></td><td></td><td></td></tr><tr><td>C57BL/6 mice</td><td>Charles River Laboratories</td><td>STRAIN CODE: 027&amp;nbsp;</td><td>C57BL/6NCrl is the exact strain we use</td></tr><tr><td>Anesthesia Dish, PYREX&amp;trade; Crystallizing Dish</td><td>Corning Life Sciences Glass&amp;nbsp;</td><td>3140125</td><td>Capacity: 25.03 oz. (740ml); Dia. x H: 4.92 x 2.55 in. (125 x 65mm). However, any small round glass container will work. A 2 cup capacity pyrex food storage bowl with flat bottom will also work and is much cheaper (Pyrex model number: 6017399).</td></tr><tr><td>Covered lead ring</td><td>Fisher Scientific</td><td>S90139C</td><td>Lead ring for stablizing flasks in a water bath. It is used inside the anesthesia dish.</td></tr><tr><td>Scalpel Blade #11</td><td>World Precision Instrucments, Inc.</td><td>500240</td><td></td></tr><tr><td>Small Vessel Cauterizer</td><td>Fine Science Tools</td><td>18000-00</td><td></td></tr><tr><td>Micro Hook</td><td>Fine Science Tools</td><td>10064-14</td><td></td></tr><tr><td>Vetbond Suture Glue</td><td>3M</td><td>1469SB</td><td>n-butyl cyanoacrylate adhesive</td></tr><tr><td>Lipopolysaccharide</td><td>Sigma Life Science</td><td>L4391</td><td>Lipopolysaccaride from E.coli 0111:B4, gamma irradiated</td></tr><tr><td>12 x 12 inch opaque box</td><td>Acrylic Display Manufacturing: A division of Piasa Plastics</td><td>C4022</td><td>Colored Acrylic 5-Sided Cube, 3/16&quot; Colored Acrylic, 12&quot;W x 12&quot;D x 12&quot;H;&amp;nbsp; http://www.acrylicdisplaymfg.com/html/cubes_19.html</td></tr><tr><td>Camera/camcorder</td><td>JVC</td><td>GC-PX100BUS</td><td>Any camcorder that works well in low light and can be imported and edited. We use the JVC GC-PX100 Full HD Everio Camcorder.</td></tr><tr><td>Covidien Tendersorb&amp;trade; Underpads</td><td>Kendall Healthcare Products Co</td><td>7174</td><td></td></tr><tr><td>WypAll L40</td><td>Kimberly-Clark Professional</td><td>5600</td><td>Any surface with moderate grip will do</td></tr><tr><td>Surface at 45 degree incline</td><td></td><td></td><td>We use a cardboard box.</td></tr><tr><td>Thin wire from a pipe cleaner</td><td>Creatology</td><td>M10314420</td><td>Any pipe cleaner from any craft store will work.</td></tr><tr><td>50mL conical tube</td><td>Falcon</td><td>352070</td><td></td></tr><tr><td>Fiberglass Screen Wire</td><td>New York Wire&amp;nbsp; www.lowes.com</td><td>14436</td><td>Any supplier can be used as long as their screen is 16 x 16 or 18 x 16</td></tr><tr><td>Razor blade</td><td>Fisherbrand</td><td>12-640</td><td>A wooden stick applicator or wooden part of a cotton-tipped swab will also work.</td></tr><tr><td>OPTIX 24-in x 4-ft x 0.22-in Clear Acrylic Sheet to make Clear Acrylic Walkway</td><td>PLASKOLITE INC</td><td>1AG2196A</td><td>Clear acrylic (1/8&quot; thick) with sides and a top to limit exploration. We bought a sheet of acrylic from a local hardware store and had them cut it to size. (2) 2&quot; x 2&quot;; (3) 2&quot; x 18&quot;; (1) 2&quot; x 15.5&quot;; (1) 2&quot; x 3&quot;. Using clear tape, tape all sides together, with the 15.5&quot; piece on top. Tape the 3&quot; piece to the end of the 15.5&quot; piece to create a flap/entryway for the mice. Alternatively, part or all of the walkway can be glued together, and only taping on the top pieces. This design will allow for the walkway to be opened for easy cleaning.</td></tr><tr><td>Protractor</td><td>Westscott</td><td>ACM14371</td><td></td></tr></tbody></table>

a battery of motor tests in a neonatal mouse model of cerebral palsy

As the sheer number of transgenic mice strains grow and rodent models of pediatric disease increase, there is an expanding need for a comprehensive, standardized battery of neonatal mouse motor tests. These tests can validate injury or disease models, determine treatment efficacy and/or assess motor behaviors in new transgenic strains. This paper presents a series of neonatal motor tests to evaluate general motor function, including ambulation, hindlimb foot angle, surface righting, negative geotaxis, front- and hindlimb suspension, grasping reflex, four limb grip strength and cliff aversion. Mice between the ages of post-natal day 2 to 14 can be used. In addition, these tests can be used for a wide range of neurological and neuromuscular pathologies, including cerebral palsy, hypoxic-ischemic encephalopathy, traumatic brain injury, spinal cord injury, neurodegenerative diseases, and neuromuscular disorders. These tests can also be used to determine the effects of pharmacological agents, as well as other types of therapeutic interventions. In this paper, motor deficits were evaluated in a novel neonatal mouse model of cerebral palsy that combines hypoxia, ischemia and inflammation. Forty-eight hours after injury, five tests out of the nine showed significant motor deficits: ambulation, hindlimb angle, hindlimb suspension, four limb grip strength, and grasping reflex. These tests revealed weakness in the hindlimbs, as well as fine motor skills such as grasping, which are similar to the motor deficits seen in human cerebral palsy patients.

小鼠用不同的小鼠每个时间点从P7（24小时以下手术）到P13（1周手术后），进行测试，以便学习测试范例不是混杂变量。 P8被选为代表性结果，小鼠显示出最大的缺陷在该时间点。 从爬行到步行转型CP新生小鼠被延迟 人类CP患者有步态异常，从脚趾走路到步态剪下。由于这种模式CP步态显示与人类相似的赤字，下床活动进行了评估。小鼠在一条直线走了得分步态对称性和肢体，爪子运动。在48小时以下的手术（PND 8），CP小鼠少对称的肢体运动和"爬行"步态相比，他们的同行假的（平均得分下床活动:CP 1.083±0.6337，N = 12 VS假1.639±0.4859，N = 9; P &lt;0.05， 图10）。一个星期，两个CP和假小鼠已经转变到正（数据未示出）。 <img alt="图10" src="/files/ftp_upload/53569/53569fig10.jpg" /> 图10. CP的乳鼠不走动以及沙姆斯。深水小鼠（黑条）有1.639±0.4859（9例），这意味着它们的动态发展不对称的肢体运动和缓慢爬行介于一个平均分。 CP小鼠（灰色条）收到的1.083±0.6337（N = 12）的平均分，这意味着他们行走欠发达，而且往往有不对称的肢体运动。数据表示为平均值±SEM; *为P &lt;0.05。 <a href="https://jove.unicartagenaproxy.elogim.com/files/ftp_upload/53569/53569fig10large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> 轩dlimb脚角度以增加CP 除了下地活动，后肢足角度进行了评估。八天的假乳鼠走他们的后爪朝前，相比HIL老鼠，走在一条直线（ 图2时，谁也张开后爪;平均角度:CP 77.48±9.848，N = 9，与对照组分别54.54± 8.043，N = 11; p &lt;0.0001， 图11）。这增加了角步态不稳相关，在幼崽需要增加他们的后爪的角度，以稳定他们的步态和平衡和协调帮助。 <img alt="图11" src="/files/ftp_upload/53569/53569fig11.jpg" /> 图11. CP的乳鼠张开他们的后爪行走时，CP鼠（黑网吧）有77.48±8.043（N = 11），它们的后肢之间的平均角度，而假小鼠（灰色B<html> ARS）有54.54±9.848（平均角N = 9）。数据表示为平均值±SEM; ****为p &lt;0.0001。 <a href="https://jove.unicartagenaproxy.elogim.com/files/ftp_upload/53569/53569fig11large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> CP小鼠没有表现出赤字时，表面正位 被列入表面扶正试验一些CP患者受损主干管（Heyrman 等 ，2013）。此外，前庭系统是必要的，以检测用于扶正的必要性，并有在一些CP患者23前庭赤字。相比假控件扶正时的CP小鼠不显示显著缺陷（数据未显示）。 CP小鼠执行相同的深水中负趋地性测试 T"FO:保together.within页="1"&gt;负趋地是用来测试在年轻的幼崽运动协调小鼠被质疑正对倾斜表面面对下坡的地方延迟或未能东方上坡可能表明赤字。在协调性，平衡，或前庭输入。当与相比，假小鼠（数据未示出）的负趋地性攻击的CP小鼠显示无缺陷。此外，CP小鼠没有表现出偏爱转向一侧与另一当重新定向。 前悬挂肢体测试适用于小鼠超过10天以上 CP患者的精细动作技能，如抓肌张力下降和赤字。为了测试在此小鼠模型中的弱点，我们使用了一个前肢悬浮试验。此外，这款机型采用单方面的缺血性损伤和片面的烦躁可能使用这种悬挂试验确定。该测试对于超过10天15老年小鼠更好。在过去8天，损伤后的两天，有CP和假小鼠之间无显著差异（数据未显示）。 后肢力量在CP减少小鼠 人类CP患者常需要大括号或辅助行走装置，由于缺乏电机控制和力量。为了比较的啮齿动物的CP模型对人类，后肢强度用后肢悬浮试验评估。当从锥形管的侧悬挂，CP小鼠显示后肢无力，如通过减少在吊得分证实（后肢悬挂得分:CP 3.468±0.5561，每组13，与假3.891±0.1329，每组13; P &lt; 0.05， 图 12）。在后肢悬挂时间没有观察到差异（数据未显示）。因此，类似于人类CP患者，CP小鼠展示出hindlIMB（腿）的弱点。 <img alt="图12" src="/files/ftp_upload/53569/53569fig12.jpg" /> 图12.深水小鼠轻微，但在他们的后肢比CP小鼠明显强。在平均得分挂的3.891±0.1329（N = 13），假鼠（黑条）表现出更多的后肢分离，因此，更强的姿态后肢，平均挂得分3.468±0.5561（N = 13）挂在管比CP小鼠（灰色条）边缘的时候。数据表示为平均值±SEM; *为P &lt;0.05。 <a href="https://jove.unicartagenaproxy.elogim.com/files/ftp_upload/53569/53569fig12large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> 握力降低的关注CP损伤 用四只爪子抓是用于在啮齿动物重要攀爬并在不平的地面上运行的条款。握要求显著持续强劲，而不是灵巧或线性力，主要是在数字和爪子24。小鼠必须持有一个倒置的丝网屏幕上自己的体重。 CP小鼠无法维持他们的抓地力和这些小鼠落到了实处显著较低的角度（4肢平均角度:CP 75.627±24.48，N = 11，与对照组分别96.57±10.836，N = 9，P &lt;0.05， 图13）。该数据表明，有在CP小鼠握力一个显著赤字。 <img alt="图13" src="/files/ftp_upload/53569/53569fig13.jpg" /> 图13. CP小鼠比沙姆斯较弱的抓地力。深水小鼠（黑条）可以把握的平均倒角96.57±10.836（N = 9）。 CP小鼠（灰色条）只能达到75.627±24.48（N = 11）倒角度。数据表示为平均值±SEM; *为p0; 0.05。 <a href="https://jove.unicartagenaproxy.elogim.com/files/ftp_upload/53569/53569fig13large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> 抓握反射赤字在CP表观小鼠 随着粗大运动障碍，精细动作的动作也受损CP患者25,26。在人类的抓握反射是在出生时并消失约5 - 6个月。然而，变化的抓握反射，比如夸张的速度和抓力，未能把握，或6个月的年龄后，抓握反射的再度出现，都表明神经系统的损害。为了比较在CP模式抓，抓反身赤字进行了测定。 在伤后48小时，CP小鼠展示抓握反射减少（平均爪子抓住在48小时:CP 2.429±0.9376，N = 14，与对照组分别3.214±0.8018，N = 14; P &lt;0.05， 图14A）。有在前爪右爪偏好轻微，但不显著增加（数据未显示）。有一个在后爪一个显著右爪偏好（CP 75.0±42.74，N = 14，与对照组分别17.86±54.09，N = 14，P &lt;0.005， 图14B）。伤后一周，CP小鼠显示抓赤字（平均爪子抓住了在第1周:CP 2.75±1.035，N = 8，与对照组分别3.80±0.6325，N = 10，P &lt;0.05， 图14C），没有显着的爪子偏好。 <img alt="图14" src="/files/ftp_upload/53569/53569fig14.jpg" /> 图14. CP小鼠有把握赤字，在后爪，对侧脑损伤区域。 （A）48小时以下的损伤（PND 8），CP小鼠（灰色条）抓住了棍子，平均比假灵魂爪子少LS（黑条）。（B）CP小鼠（灰色条）显示的右后爪（对侧受伤），而不是使用左后肢（同侧伤害）抓的偏好。深水小鼠（黑条）不显示此右爪偏好。右爪子偏好的计算公式为（[右爪子-左爪子] / [右爪子+左爪子+两个爪子] * 100）（C）一周损伤后，CP小鼠（灰色条）仍然显示抓赤字相比。沙姆斯（黑条）。数据表示为平均值±SEM; *为p &lt;0.05，**是P &lt;0.005。 <a href="https://jove.unicartagenaproxy.elogim.com/files/ftp_upload/53569/53569fig14large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> CP小鼠转远离边缘在悬崖厌恶 悬崖厌恶测试依赖于小鼠转AWA的内在恐惧y坐标从对安全陡峭的山崖和头部。虽然一些CP患者有前庭的困难，以及受损马达控制，在CP小鼠没有显示在此测试的任何缺陷。

Watch this Scientific Journal Video about A Battery of Motor Tests in a Neonatal Mouse Model of Cerebral Palsy at JoVE.com

A Battery of Motor Tests in a Neonatal Mouse Model of Cerebral Palsy

Presented is a concise battery of mouse neonatal motor tests. Using these tests, neonatal motor deficits can be demonstrated in a variety of neonatal motor disorders. By having a standardized set of tests, results from different studies can be compared, allowing for better and accurate reporting between groups.

汽车测试的脑瘫新生儿小鼠模型电池

As the sheer number of transgenic mice strains grow and rodent models of pediatric disease increase, there is an expanding need for a comprehensive, ...

a-battery-of-motor-tests-in-a-neonatal-mouse-model-of-cerebral-palsy

Universidad de Cartagena UDEC

Research

JoVE Journal

Medicine

24.0K 次观看.  Lewiz Katz School of Medicine at Temple University. Presented is a concise battery of mouse neonatal motor tests. Using these tests, neonatal motor deficits can be demonstrated in a variety of neonatal motor disorders. By having a standardized set of tests, results from different studies can be compared, allowing for better and accurate reporting between groups.

视频: A Battery of Motor Tests in a Neonatal Mouse Model of Cerebral Palsy

Assessment of Sensorimotor Function in Mouse Models of Parkinson's Disease

Sensitive and reliable behavioral outcome measures are essential to the evaluation of potential therapeutic treatments in preclinical trials for many neurodegenerative diseases. In Parkinson's disease, sensorimotor tests sensitive to varying degrees of nigrostriatal dysfunction are fundamental for testing the efficacy of potential therapeutics. Reliable and quite elegant sensorimotor measures exist for rats, however many of these tests measure sensorimotor asymmetry within the rat and are not entirely suitable for the newer genetic mouse models of PD. We have put together a battery of sensorimotor tests inspired by the sensitive tests in rats and adapted for mice. The test battery highlighted in this study is chosen for a) its sensitivity in a wide variety of mouse models of PD, b) its ease in implementing into a study, and c) its low expense. These tests have proven useful in characterizing novel genetic mouse models of PD as well as in testing potential disease-modifying therapies.

In Parkinson's disease and movement disorders in general, sensitive and reliable behavioral assays are essential for testing novel potential therapeutics. Here, we describe a manageable battery of sensorimotor tests for mice that are sensitive to varying degrees of injury to the nigrostriatal system and useful for preclinical studies.

帕金森氏症小鼠模型的感觉功能评估

Sensitive and  reliable behavioral outcome measures are essential to the evaluation of  potential therapeutic treatments in preclinical trials for many  ...

科研

行为学

Paw-Dragging: a Novel, Sensitive Analysis of the Mouse Cylinder Test

The cylinder test is routinely used to predict focal ischemic damage to the forelimb motor cortex in rodents. When placed in the cylinder, rodents explore by rearing and touching the walls of the cylinder with their forelimb paws for postural support. Following ischemic injury to the forelimb sensorimotor cortex, rats rely more heavily on their unaffected forelimb paw for postural support resulting in fewer touches with their affected paw which is termed forelimb asymmetry. In contrast, focal ischemic damage in the mouse brain fails to result in comparable consistent deficits in forelimb asymmetry. While forelimb asymmetry deficits are infrequently observed, mice do demonstrate a novel behaviour post stroke termed &#8220;paw-dragging&#8221;. Paw-dragging is the tendency for a mouse to drag its affected paw along the cylinder wall rather than directly push off from the wall when dismounting from a rear to a four-legged stance. We have previously demonstrated that paw-dragging behaviour is highly sensitive to small cortical ischemic injuries to the forelimb motor cortex. Here we provide a detailed protocol for paw-dragging analysis. We define what a paw-drag is and demonstrate how to quantify paw-dragging behaviour. The cylinder test is a simple and inexpensive test to administer and does not require pre-training or food deprivation strategies. In using paw-dragging analysis with the cylinder test, it fills a niche for predicting cortical ischemic injuries such as photothrombosis and Endothelin-1 (ET-1)-induced ischemia &#8211; two models that are ever-increasing in popularity and produce smaller focal injuries than middle cerebral artery occlusion. Finally, measuring paw-dragging behaviour in the cylinder test will allow studies of functional recovery after cortical injury using a wide cohort of transgenic mouse strains where previous forelimb asymmetry analysis has failed to detect consistent deficits.

Classical forelimb asymmetry analysis of the cylinder test is routinely used to assess behavioural deficits in rats following brain injury or stroke; however, it fails to detect consistent deficits in mice. This study demonstrates that quantifying paw-dragging behaviour is a more sensitive analysis of brain injury in mice.

PAW-拖动:一本小说，鼠标缸试验敏感性分析

The cylinder test is routinely used to predict focal ischemic damage to the forelimb motor cortex in rodents. When placed in the cylinder, rodents explore ...

Measuring Motor Coordination in Mice

Mice are increasingly being used in behavioral neuroscience, largely replacing rats as the behaviorist's animal of choice. Before aspects of behavior such as emotionality or cognition can be assessed, however, it is vital to determine whether the motor capabilities of e.g. a mutant or lesioned mouse allow such an assessment. Performance on a maze task requiring strength and coordination, such as the Morris water maze, might well be impaired in a mouse by motor, rather than cognitive, impairments, so it is essential to selectively dissect the latter from the former. For example, sensorimotor impairments caused by NMDA antagonists have been shown to impair water maze performance2. 
Motor coordination has traditionally been assessed in mice and rats by the rotarod test, in which the animal is placed on a horizontal rod that rotates about its long axis; the animal must walk forwards to remain upright and not fall off. Both set speed and accelerating versions of the rotarod are available.
The other three tests described in this article (horizontal bar, static rods and parallel bars) all measure coordination on static apparatus. The horizontal bar also requires strength for adequate performance, particularly of the forelimbs as the mouse initially grips the bar just with the front paws. 
Adult rats do not perform well on tests such as the static rods and parallel bars (personal observations); they appear less well coordinated than mice. I have only tested male rats, however, and male mice seem generally less well coordinated than females. Mice appear to have a higher strength:weight ratio than rats; the Latin name, Mus musculus, seems entirely appropriate. The rotarod, the variations of the foot fault test12 or the Catwalk (Noldus)15 apparatus are generally used to assess motor coordination in rats.

Protocols are presented for two established motor coordination tasks, the accelerating rotarod and horizontal bar, also two tests developed in Oxford recently, the static rods and parallel bars. These tests can detect motor impairments potentially of interest in their own right, as well as being possible variables in tests of other areas of behavior.

测量小鼠运动协调

Mice are increasingly being used in behavioral neuroscience, largely replacing rats as the behaviorist's animal of choice. Before aspects of behavior such ...

神经科学

Neurodevelopmental Reflex Testing in Neonatal Rat Pups

Neurodevelopmental reflex testing is commonly used in clinical practice to assess the maturation of the nervous system. Neurodevelopmental reflexes are also referred to as primitive reflexes. They are sensitive and consistent with later outcomes. Abnormal reflexes are described as an absence, persistence, reappearance, or latency of reflexes, which are predictive indices of infants that are at high risk for neurodevelopmental disorders. Animal models of neurodevelopmental disabilities, such as cerebral palsy, often display aberrant developmental reflexes, as would be observed in human infants. The techniques described assess a variety of neurodevelopmental reflexes in neonatal rats. Neurodevelopmental reflex testing offers the investigator a testing method that is not otherwise available in such young animals. The methodology presented here aims to assist investigators in examining developmental milestones in neonatal rats as a method of detecting early-onset brain injury and/or determining the effectiveness of therapeutic interventions. The methodology presented here aims to provide a general guideline for investigators.

Behavioral testing is the gold standard for determining outcomes following brain injury, and can identify the presence of developmental disabilities in infants and children. Neurodevelopmental reflexes are an early indicator of these abnormalities. A host of easily accomplished developmental reflex tests in the neonatal rodent were developed and described here.

神经发育复检新生幼鼠

Neurodevelopmental reflex testing is commonly used in clinical practice to assess the maturation of the nervous system. Neurodevelopmental reflexes are ...

Battery of Behavioral Tests Assessing General Locomotion, Muscular Strength, and Coordination in Mice

Behavioral testing is used in pre-clinical trials to assess the phenotypic effects and outcomes that a particular disease or treatment has on the animal&#39;s wellbeing and health. There are numerous behavioral tests that may be applied. We selected a test for general locomotion, the open field test (OFT); a test for muscular strength, the mesh test (MT); and a test for coordination, the rotarod test (RR). Testing can be accomplished on a weekly or monthly basis. As a test for general locomotion, the OFT works by objectively monitoring movement parameters while the mouse is in an open field apparatus. The field is generally a 2&#39; x 2&#39; box, and the movements are recorded through laser sensing or through video capture. The mouse is placed in the center of the open field and allowed to move freely for the test. The MT uses the latency for a mouse to fall off an inverted screen as a measure of muscular strength. A mouse is placed on a screen, which is inverted over a clear box, and is timed for their latency to fall. Three trials are performed, with the best of the three trials scored for that day. A score of 60 s is the maximum time a mouse is left inverted. Mice are given a 5-min rest period between mesh test trials. Lastly, an accelerated protocol on the RR assesses motor coordination and endurance. During a trial, a mouse walks on a rotating rod as it increases in speed from 4 rpm to 40 rpm over 5 min. The trial ends when the mouse touches the magnetized pressure sensor upon falling. Each mouse undergoes three trials, and the best trial is scored for that day. This combined behavioral data allows for the global assessment of mobility, coordination, strength, and movement of the test animals. At least two out of the three behavioral testing measures must show improvement for an animal to qualify as having overall improved motor function.

Behavioral testing is used in pre-clinical trials to assess the phenotypic effects of a disease or treatment on an animal's wellbeing. In order to globally assess motor functioning, we selected tests for general locomotion, muscular strength, and coordination: the open field test, the mesh test, and the rotarod test, respectively.

用于评估一般运动、肌肉力量和小鼠协调性的行为测试电池

Behavioral testing is used in pre-clinical trials to assess the phenotypic effects and outcomes that a particular disease or treatment has on the ...

A Neonatal Mouse Spinal Cord Compression Injury Model

Spinal cord injury (SCI) typically causes devastating neurological deficits, particularly through damage to fibers descending from the brain to the spinal cord. A major current area of research is focused on the mechanisms of adaptive plasticity that underlie spontaneous or induced functional recovery following SCI. Spontaneous functional recovery is reported to be greater early in life, raising interesting questions about how adaptive plasticity changes as the spinal cord develops. To facilitate investigation of this dynamic, we have developed a SCI model in the neonatal mouse. The model has relevance for pediatric SCI, which is too little studied. Because neural plasticity in the adult involves some of the same mechanisms as neural plasticity in early life1, this model may potentially have some relevance also for adult SCI. Here we describe the entire procedure for generating a reproducible spinal cord compression (SCC) injury in the neonatal mouse as early as postnatal (P) day 1. SCC is achieved by performing a laminectomy at a given spinal level (here described at thoracic levels 9-11) and then using a modified Yasargil aneurysm mini-clip to rapidly compress and decompress the spinal cord. As previously described, the injured neonatal mice can be tested for behavioral deficits or sacrificed for ex vivo physiological analysis of synaptic connectivity using electrophysiological and high-throughput optical recording techniques1. Earlier and ongoing studies using behavioral and physiological assessment have demonstrated a dramatic, acute impairment of hindlimb motility followed by a complete functional recovery within 2 weeks, and the first evidence of changes in functional circuitry at the level of identified descending synaptic connections1.

This article describes a method for generating a reproducible spinal cord compression injury (SCI) in the neonatal mouse. The model provides an advantageous platform for studying mechanisms of adaptive plasticity that underlie spontaneous functional recovery.

新生儿小鼠脊髓压迫损伤模型

Spinal cord injury (SCI) typically causes devastating neurological deficits, particularly through damage to fibers descending from the brain to the spinal ...

医学

Neurobehavioral Assessments in a Mouse Model of Neonatal Hypoxic-ischemic Brain Injury

We performed unilateral carotid artery occlusion on CD-1 mice to create a neonatal hypoxic-ischemic (HI) model and investigated the effects of neonatal HI brain injury by studying neurobehavioral functions in these mice compared to non-operated (i.e., normal) mice. During the study, Rice-Vannucci&#39;s method was used to induce neonatal HI brain damage in postnatal day 7-10 (P7-10) mice. The HI operation was performed on the pups by unilateral carotid artery ligation and exposure to hypoxia (8% O2 and 92% N2 for 90 min). One week after the operation, the damaged brains were evaluated with the naked eye through the semi-transparent skull and were categorized into subgroups based on the absence (&#34;no cortical injury&#34; group) or presence (&#34;cortical injury&#34; group) of cortical injury, such as a lesion in the right hemisphere. On week 6, the following neurobehavioral tests were performed to evaluate the cognitive and motor functions: passive avoidance task (PAT), ladder walking test, and grip strength test. These behavioral tests are helpful in determining the effects of neonatal HI brain injury and are used in other mouse models of neurodegenerative diseases. In this study, neonatal HI brain injury mice showed motor deficits that corresponded to right hemisphere damage. The behavioral test results are relevant to the deficits observed in human neonatal HI patients, such as cerebral palsy or neonatal stroke patients. In this study, a mouse model of neonatal HI brain injury was established and showed different degrees of motor deficits and cognitive impairment compared to non-operated mice. This work provides basic information on the HI mouse model. MRI images demonstrate the different phenotypes, separated according to the severity of brain damage by motor and cognitive tests.

We performed unilateral carotid artery occlusion on postnatal day 7-10 CD-1 mouse pups to create a neonatal hypoxic-ischemic (HI) model and investigated the effects of HI brain injury. We studied neurobehavioral functions in these mice compared to non-operated normal mice.

小鼠新生儿缺氧缺血性脑损伤模型的神经行为评估

We performed unilateral carotid artery occlusion on CD-1 mice to create a neonatal hypoxic-ischemic (HI) model and investigated the effects of neonatal HI ...

A Controlled Mouse Model for Neonatal Polymicrobial Sepsis

Neonatal sepsis remains a global burden. A preclinical model to screen effective prophylactic or therapeutic interventions is needed. Neonatal mouse polymicrobial sepsis can be induced by injecting cecal slurry intraperitoneally into day of life 7 mice and monitoring them for the following week. Presented here are the detailed steps necessary for the implementation of this neonatal sepsis model. This includes making a homogeneous cecal slurry stock, diluting it to a weight- and litter-adjusted dose, an outline of the monitoring schedule, and a definition of observed health categories used to define humane endpoints. The generation of a homogeneous cecal slurry stock from pooled donors allows for the administration into many litters over time, reducing the variation between donors, and preventing the use of potentially toxic glycerol. The monitoring strategy used allows for the anticipation of survival outcome and the identification of mice that would later progress to death, allowing for an earlier identification of the humane endpoint. Two main behavioral features are used to define the health scores, namely, the ability of the neonatal mice to right themselves when placed on their back and their level of mobility. These criteria could potentially be applied to address humane endpoints in other studies of neonatal disease in mice, as long as a pilot study is performed to confirm accuracy. In conclusion, this approach provides a standardized method to model newborn sepsis in mice, while providing resources to assess animal welfare used to define early humane endpoints for challenged animals.

This protocol provides the necessary steps to establish and evaluate neonatal sepsis in 7-day-old mice.

新生儿多药脓毒症的对照小鼠模型

Neonatal sepsis remains a global burden. A preclinical model to screen effective prophylactic or therapeutic interventions is needed. Neonatal mouse ...

免疫与感染

A Mouse Model of Single and Repetitive Mild Traumatic Brain Injury

Mild Traumatic Brain Injury (mTBI) can result in the acute loss of brain function, including a period of confusion, a loss of consciousness (LOC), focal neurological deficits and even amnesia. Athletes participating in contact sports are at high risk of exposure to large number of mTBIs. In terms of the level of injury in a sporting athlete, a mTBI is defined as a mild injury that does not cause gross pathological changes, but does cause short-term neurological deficits that are spontaneously resolved. Despite previous attempts to model mTBI in mice and rats, many have reported gross adverse effects including skull fractures, intracerebral bleeding, axonal injury and neuronal cell death. Herein, we describe our highly reproducible animal model of mTBI that reproduces clinically relevant symptoms. This model uses a custom made pneumatic impactor device to deliver a closed-head trauma. This impact is made under precise velocity and deformation parameters, creating a reliable and reproducible model to examine the mechanisms that contribute to effects of single or repetitive concussive mTBI.

Athletes absorb several hundred mild traumatic brain injuries (mTBI)/concussions every year; however, the consequence of these on the brain is poorly understood. Therefore, an animal model of single and repetitive mTBI that consistently replicates clinically relevant symptoms provides the means to advance the study of mTBI and concussion.

单一和重复轻度创伤性脑损伤的小鼠模型

Mild Traumatic Brain Injury (mTBI) can result in the acute loss of brain function, including a period of confusion, a loss of consciousness (LOC), focal ...

使用 MouseWalker 量化脊髓损伤小鼠的运动功能障碍

Using the MouseWalker to Quantify Locomotor Dysfunction in a Mouse Model of Spinal Cord Injury

The execution of complex and highly coordinated motor programs, such as walking and running, is dependent on the rhythmic activation of spinal and supra-spinal circuits. After a thoracic spinal cord injury, communication with upstream circuits is impaired. This, in turn, leads to a loss of coordination, with limited recovery potential. Hence, to better evaluate the degree of recovery after the administration of drugs or therapies, there is a necessity for new, more detailed, and accurate tools to quantify gait, limb coordination, and other fine aspects of locomotor behavior in animal models of spinal cord injury. Several assays have been developed over the years to quantitatively assess free-walking behavior in rodents; however, they usually lack direct measurements related to stepping gait strategies, footprint patterns, and coordination. To address these shortcomings, an updated version of the MouseWalker, which combines a frustrated total internal reflection (fTIR) walkway with tracking and quantification software, is provided. This open-source system has been adapted to extract several graphical outputs and kinematic parameters, and a set of post-quantification tools can be to analyze the output data provided. This manuscript also demonstrates how this method, allied with already established behavioral tests, quantitatively describes locomotor deficits following spinal cord injury.

作者聚焦:使用 MouseWalker 量化脊髓损伤小鼠模型中的运动功能障碍  

An experimental pipeline to quantitatively describe the locomotor pattern of freely walking mice using the MouseWalker (MW) toolbox is provided, ranging from initial video recordings and tracking to post-quantification analysis. A spinal cord contusion injury model in mice is employed to demonstrate the usefulness of the MW system.