提交人感谢帕特里夏·沃德博士、4月赫里蒂博士和苏珊·哈克马博士的投入和指导, 克里斯汀·雅伯里的手术协助, 陈阳生, 安德烈·维尔海特和约翰尼·莫豪斯, 感谢他们的技术援助。与统计和行为评估。国防部 (w81xwh-11-1-0668 和 w81xwh-15-15-1-0656) 和肯塔基州脊髓和头部伤害研究信托基金 (kscirt 14-5) 为这项工作提供了资金支持。

所述的所有方法都已得到路易斯维尔大学动物护理和使用机构委员会 (iacuc) 的批准。
1. 损伤前处理和检测 (sci 前一周)
<ol><li>每天处理每只老鼠 5-10分钟, 每天一次, 为期5天。 注: 本协议中使用的是最初年龄约50天、体重200-225 克的成年雄性 wistar 大鼠。在受伤前的时间点上的老鼠没有适应 lt 使用的线束, 因为充分使用后肢可以让老鼠从夹克中逃脱。</li>
	<li>进行任何特定于研究的损伤前测试 (例如, 作者对涉及 sci 对膀胱和肠道功能影响的研究进行代谢笼评估)。</li>
</ol>2. 脊髓连接25、26、27、28
<ol><li>根据提供的剂量图 (表1), 用氯胺酮 (80 mgkg) 和 xylazine (10mg源于 kg) 在腹腔内对动物进行麻醉。根据需要管理补充剂量。通过评估角膜、眼睑、踏板、尾部夹紧和羽鼻反射, 至少每10分钟测试一次麻醉深度。</li>
	<li>从发生切口和损伤的动物背部剪发。用皮草4% 手术擦洗清洁手术区域。使用长效普通抗生素 (例如, 0.5 cc 亲 pen-g 皮下注射)。</li>
	<li>将麻醉后的动物放置在较低的加热垫上, 以保持正常体温。</li>
	<li>根据椎体突起和 #10 手术刀估计目标病变水平的位置, 在动物的背上进行估计5厘米的切口, 直接在中线椎体上方。</li>
	<li>对于胸腔中挫伤,通过切除 (含 t7 椎板) 的 t8t9 水平的脊髓。</li>
	<li>使用挫伤装置, 如无限地平线撞击器29, 执行挫伤 (对于中等至严重程度的 sci, 使用 210 kdyn 的力量, 没有停留时间)18。</li>
	<li>使用4-0 直径的单丝将脊髓上的肌层和筋膜缝合在一起, 并用9毫米手术伤口缝合皮肤。</li>
	<li>术后服用硫酸庆大霉素 (每天 5 mg kg, 为期 5天; 避免膀胱感染的抗生素) 和美洛昔康 (皮下1 mg/kg, 先48小时镇痛, 然后根据需要)。</li>
	<li>将动物放在干净的笼子里, 放在加热垫上。每15分钟检查一次动物生命体征, 直到它们从麻醉中完全清醒。 在术后的第一个日子里, 鼓励动物吃含糖的食物。在前 48小时 (在手工创作时每天三次----见 2.10), 对老鼠进行不活动、在处理时发声以及缺乏吃喝欲望的监测。 如果发现镇痛不足, 请与兽医人员联系。在最初的两周恢复阶段, 观察动物感染或其他并发症的证据。一旦反射排尿返回, 这些动物往往每天两次 (清晨和下午晚些时候)。感染或体重显著下降的动物立即被安乐死。关于食物和水的摄入, 安乐死的截止点是动物体重下降超过 2 0% 的时候。正常体重减轻手术后和不使用萎缩的肌肉低于受伤水平是15-20。所有动物每周至少称重一次。</li>
	<li>每天3次使用手动微管机动 (上午 8点, 下午 3点, 晚上 10点) 进行膀胱排空手术, 直到反身膀胱功能恢复 (平均3-6天的挫伤)26,30。</li>
</ol>3. 培训阶段
<ol><li>在 sci 后两周内开始 lt, 因为过早启动干预可能会加剧继发性损伤级联 31.</li>
	<li>第1周适应跑步机训练: 将老鼠运送到专门用于训练的安静房间。</li>
	<li>在第一天, 随机和均匀地将 sci 动物分为训练和未训练的对照组, 以考虑损伤本身的潜在变异性以及挫伤后的自发恢复程度。例如, 将老鼠分成4组: 四足训练 (qt)、前肢训练 (ft)、未经训练的控制 (nt) 和未训练的家庭保持架控制 (hc)。动物接受椎板切除术但没有受伤, 否则处理方式与其他组相同的假组, 也可以在没有训练的情况下作为未受伤的控制组使用。</li>
	<li>将每只动物放置在各自的线束中 (图 1), 并通过紧固在重量支撑弹簧上的鳄鱼夹将线束固定在跑步机上方的体重支撑机构上 (图2和图 3)。这就要求动物固定在跑步机上的一个位置, 确保它们朝着指定的前进方向和速度前进。 注: 由于时间和人员的限制, 作者的实验室每天对12只动物进行培训, 每个子集组有3只动物。</li>
	<li>按照以前发布的协议17开始适应过程.开始适应 lt (sci 后第3周开始), 逐步采用跑步机暴露方案, 从第1天的10分钟增加到第一周58分钟的全部目标 (表 2)。通常情况下, 到第4天, 动物很好地适应了训练方案。如果动物在适应的第三天还没有显示出进展, 时间就会缩短, 额外的天数会在更渐进的情况下增加 (罕见的情况)。<ol>
<li>如果动物在第一天或两天内不适应线束和跑步机的限制, 停止训练, 将其从马具中取出, 将动物放回笼子里, 并给予它两种食物, 以帮助加强未来的依从性。第二天, 将动物再次放置在马具和重量支撑系统中 1 0分钟。在随后的日子里, 最初将训练时间延长 20分钟, 然后继续每天增加训练时间, 以便在第10天之前完成全面培训。</li>
	</ol>
</li>
	<li>按照表 2中提供的详细培训方案进行操作。<ol>
<li>由于损伤后后肢使用有限, qt 组的大鼠在踩在跑步机上时需要人工协助进行适当的爪子放置。每只手使用一个手指 (通常是第三个数字) 来帮助支撑腰部。当动物在步进方面需要进一步的帮助时, 用同样的手指在膝盖以上施加压力, 开始踩。如有必要, 使用单独的手指 (通常是第五位数字) 来帮助脚踩。 注: 所需的体重支持量因动物而异, 并随着训练的进行而变化。弹簧支撑系统提供了足够的帮助, 使动物能够获得适当的步态。培训员根据上述要求提供进一步支助。请注意, lt 的一个关键要素是功能上适合步进和肢体间协调的爪子放置, 该位置由培训师推动, 独立于支持系统。</li>
		<li>对于 ft 运动组, 调整体重支持系统, 使后肢稍微抬起, 以确保没有感觉刺激爪子, 也不因与跑步机接触而产生负重。 注: 英国《金融时报》组作为一种运动和代谢控制, 类似于基于人类活动的培训研究中的手摇运动。</li>
		<li>以与 qt 类似的方式将 nt 组利用并连接到体重支持系统, 并将 nt 组放置在固定表面的 qt 群附近 (图2和图 3)。 注: nt 组在较长时间内被利用时没有收到任何活动和控件。</li>
		<li>家庭笼子组可以作为一个额外的控制。将这些动物运送到训练设施, 作为这一群体的额外步骤。</li>
	</ol>
</li>
	<li>在 lt 开始后的第7-10 天, 每天对每只动物进行一次训练, 直到研究结束的那一天。在每天的训练之后, 给每只动物一个含糖的治疗, 以加强依从性。在研究期间, 继续每天对动物进行 lt, 方法是遵循表 2中提供的1年方案 (例如, 8-12, 以模拟临床研究中进行的大约80小时的疗程)9。</li>
</ol>4. 安乐死和组织收集
<ol><li>对坚持 avma 安乐死指南的动物进行致命剂量的麻醉。</li>
	<li>当心脏刚刚跳动时, 立即开始在专用的烟罩中使用冷的肝素盐, 然后是冷的、4% 的甲醛溶液, 以此为原料。<ol>
<li>开始使用手术剪刀在横隔膜上做一个切口, 露出胸腔。继续从两边的胸腔进行切割, 取出胸腔。将灌注针插入心脏的左心室, 用止血器夹住针头, 然后夹住右心房。</li>
		<li>使用灌注泵机制, 让冷肝素盐水流经动物的血管。一旦清盐水从右心房流出, 切换到冷的4% 的甲醛溶液, 直到身体变硬。</li>
	</ol>
</li>
	<li>去除必要的组织, 如肾脏、膀胱、结肠、大脑、感觉神经节和脊髓, 并在4°c 下以4% 的甲醛储存长达48小时。24-48小时后, 将组织移动到30% 的蔗糖, 并在4°c 下保存。</li>
	<li>将收集到的组织转移到30% 的磷酸蔗糖缓冲低温保护剂溶液中, 直到组织准备好切割。要切割组织, 请将其嵌入组织冷冻化合物中, 并根据所使用的组织类型在所需厚度下切割在低温恒温器上 (例如, 大脑和脊髓组织为 35μm, 器官组织为5-7 微米)。</li>
</ol>

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H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Persistent+polyuria+in+a+rat+spinal+contusion+model.">Persistent polyuria in a rat spinal contusion model.</a> Journal of Neurotrauma. 29 (15), 2490-2498 (2012).</li><li>Scheff, S. W., Rabchevsky, A. G., Fugaccia, I., Main, J. A., Lumpp, J. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Experimental+modeling+of+spinal+cord+injury:+characterization+of+a+force-defined+injury+device.">Experimental modeling of spinal cord injury: characterization of a force-defined injury device.</a> Journal of Neurotrauma. 20 (2), 179-193 (2003).</li><li>Ferrero, S. L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effects+of+lateral+funiculus+sparing,+spinal+lesion+level,+and+gender+on+recovery+of+bladder+voiding+reflexes+and+hematuria+in+rats.">Effects of lateral funiculus sparing, spinal lesion level, and gender on recovery of bladder voiding reflexes and hematuria in rats.</a> Journal of Neurotrauma. 32 (3), 200-208 (2015).</li><li>Smith, R. R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Swim+training+initiated+acutely+after+spinal+cord+injury+is+ineffective+and+induces+extravasation+in+and+around+the+epicenter.">Swim training initiated acutely after spinal cord injury is ineffective and induces extravasation in and around the epicenter.</a> Journal of Neurotrauma. 26 (7), 1017-1027 (2009).</li><li>Oelke, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+practical+approach+to+the+management+of+nocturia.">A practical approach to the management of nocturia.</a> International Journal of Clinical Practice. 71 (11), e13027 (2017).</li><li>Claydon, V., Steeves, J., Krassioukov, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Orthostatic+hypotension+following+spinal+cord+injury:+understanding+clinical+pathophysiology.">Orthostatic hypotension following spinal cord injury: understanding clinical pathophysiology.</a> Spinal Cord. 44 (6), 341 (2006).</li><li>Antunes-Rodrigues, J., De Castro, M., Elias, L. L., Valenca, M. M., McCANN, S. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neuroendocrine+control+of+body+fluid+metabolism.">Neuroendocrine control of body fluid metabolism.</a> Physiological Reviews. 84 (1), 169-208 (2004).</li><li>C&#244;t&#233;, M. -. P., Azzam, G. A., Lemay, M. A., Zhukareva, V., Houl&#233;, J. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Activity-dependent+increase+in+neurotrophic+factors+is+associated+with+an+enhanced+modulation+of+spinal+reflexes+after+spinal+cord+injury.">Activity-dependent increase in neurotrophic factors is associated with an enhanced modulation of spinal reflexes after spinal cord injury.</a> Journal of Neurotrauma. 28 (2), 299-309 (2011).</li><li>Dupont-Versteegden, E. E., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Exercise-induced+gene+expression+in+soleus+muscle+is+dependent+on+time+after+spinal+cord+injury+in+rats.">Exercise-induced gene expression in soleus muscle is dependent on time after spinal cord injury in rats.</a> Muscle &amp; Nerve: Official Journal of the American Association of Electrodiagnostic Medicine. 29 (1), 73-81 (2004).</li><li>De Leon, R., Hodgson, J., Roy, R., Edgerton, V. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Full+weight-bearing+hindlimb+standing+following+stand+training+in+the+adult+spinal+cat.">Full weight-bearing hindlimb standing following stand training in the adult spinal cat.</a> Journal of Neurophysiology. 80 (1), 83-91 (1998).</li><li>Pearson, K. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Progress in brain research. 143, 123-129 (2004).</li><li>Gerasimenko, Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Feed-forwardness+of+spinal+networks+in+posture+and+locomotion.">Feed-forwardness of spinal networks in posture and locomotion.</a> The Neuroscientist. 23 (5), 441-453 (2017).</li><li>Courtine, G., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Transformation+of+nonfunctional+spinal+circuits+into+functional+states+after+the+loss+of+brain+input.">Transformation of nonfunctional spinal circuits into functional states after the loss of brain input.</a> Nature Neuroscience. 12 (10), 1333 (2009).</li></ol>

作者没有什么可透露的。

sci 后 abt 对大鼠的治疗方法是一种新的治疗干预方法。虽然动物模型中可能存在其他运动和阶梯式训练方法,但这种方法模仿了 sci 人群中临床进行的 lt, 我们在那里看到了有希望的结果23。结合我们对控制动物的设置、方案和使用, 利用我们的训练范式所获得的结果将有助于了解 sci 之后 abt 的好处. 本协议的未来应用包括观察所描述的结果:abt 在不同的训练时间框架, 以及 abt 对恢复从不同的水平和程度的伤害的影响。
这种设计的一个局限性是这种实验的时间长度。鉴于我们对每只动物的训练方案每天需要 1小时, 10周每天每天都需要大量的人员时间和有组织的时间表是必要的。需要特别注意的一个重要方面是英国《金融时报》集团, 该集团拥有独特的线束, 带有钩环材料带, 以固定跑步机上方的后肢, 从而消除体重支撑。重要的是要确保动物不会得到重量支持, 这也是为什么一个平台没有被放置在老鼠的后爪下的原因。此外, 正如以前的研究表明, 感觉输入是脊髓运动系统可塑性的主要驱动因素 38,39, 40, 不断需要处理 qt 组, 以协助在临床环境中与理疗师的步子大同小异。
对用于动物的商用跑步机系统进行的一项重要修改是扭转极性。暴露电机后, 正线和负线被切换, 扭转了跑步机的运动方向。这使得更多的空间和更容易进入和帮助训练的动物 (该系统配备了一个冲击网格在一端, 旨在防止未经利用的, 空间完整的动物从跑步机带上走下来)。

在全球范围内, 有 250, 000 至 500, 000 例新的脊髓损伤 (sci) 病例要么因退化、疾病, 要么最常见的 (高达 90%) 创伤1。创伤性 sci 后, 一系列生理事件发生, 导致神经功能缺陷, 影响多种身体功能。由于 sci 之后的长期缺陷, 制定和测试有效的治疗方式至关重要。直到最近, 康复战略最普遍的重点是恢复行动能力 2、3.继 sci 之后, 患者将泌尿系统、肠道和性功能列为生活并发症中需要更好的管理1,4,5。因此, 从康复的角度来看, 以膀胱、肠道和性功能为目标是最重要的, 从 1,4,5。
运动和运动训练 (lt) 是 sci 患者群体常用的康复疗法, 具有心血管功能、泌尿系统功能和行动能力 6、7、8等诸多优点 ,9,10。正因为如此, 我们在临床前大鼠 sci 模型中采用了类似的模式。我们的目标是确定 lt 对 sci wistar 大鼠有什么影响, 特别是对上 (肾) 和下 (膀胱, 外尿道括约肌) 尿路功能、肠功能和性功能。此外, lt 已被证明足以激活低于损伤水平的神经肌肉系统, 这可能会影响中枢神经系统 (cns)内的可塑性 11,12。
lt 在临床前研究中的成功在 13、14和小型15、16、17、18、19 sci 动物模型中都有很好的记录。有证据表明, lt 提供的传入感官输入足以刺激脊柱反射通路, 从而导致可塑性和改善感觉-运动功能9,20。关于自主功能的 lt 福利还没有很好的特征。为此, 我们实施我们的培训范式, 重点是自主结果测量, 使用四个不同的组, 包括两个未受过训练的控制和代谢锻炼非重量轴承组, 同时一个 lt 组, 模仿时间,用于临床研究的会话持续时间、手动辅助和重量支持 19、21、22、23、24.

<table>
	<tbody>
		<tr>
			<td>Exer-3R treadmill</td>
			<td>Columbus Instruments</td>
			<td></td>
			<td>reversed polarity of the motor</td>
		</tr>
		<tr>
			<td>Body weight support system</td>
			<td>N/A</td>
			<td>N/A</td>
			<td>modified spring scales with alligator clips</td>
		</tr>
		<tr>
			<td>Rat harness</td>
			<td>N/A</td>
			<td>N/A</td>
			<td>Our harnesses are custom made; please refer to Figure 1 for visual.</td>
		</tr>
		<tr>
			<td>Infinite Horizon (IH) impactor device</td>
			<td>Precision Systems and Instrumentation</td>
			<td>Model 0400</td>
			<td></td>
		</tr>
		<tr>
			<td>Ketamine HCl</td>
			<td>Hospira</td>
			<td>NDC 0409-2053-10</td>
			<td></td>
		</tr>
		<tr>
			<td>Xylazine (AnaSed Injection)</td>
			<td>Akorn Animal Health</td>
			<td>NDC 59399-110-20</td>
			<td></td>
		</tr>
		<tr>
			<td>Meloxicam (Eloxiject)</td>
			<td>Henry Schein Animal Health</td>
			<td>NDC 116695-6925-2</td>
			<td></td>
		</tr>
		<tr>
			<td>Gentamicin Sulfate (GentaFuse)</td>
			<td>Henry Schein Animal Health</td>
			<td>NDC 11695-4146-1</td>
			<td></td>
		</tr>
		<tr>
			<td>urethane, 97%</td>
			<td>Argos Organics</td>
			<td>CAS 51-79-6</td>
			<td></td>
		</tr>
		<tr>
			<td>4-0 monofilament suture kit (4-0 Ethilon Nylon Suture)</td>
			<td>Ethicon, LLC</td>
			<td>205016</td>
			<td></td>
		</tr>
		<tr>
			<td>Michel suture clips (9mm Auto Clips)</td>
			<td>MikRon Precision, Inc.</td>
			<td>1629</td>
			<td></td>
		</tr>
		<tr>
			<td>Heating pad</td>
			<td>Mastex Industries, Inc</td>
			<td>Model 500</td>
			<td></td>
		</tr>
		<tr>
			<td>Tootie Fruitys cereal</td>
			<td>Malt O Meal</td>
			<td></td>
			<td>For training reward</td>
		</tr>
		<tr>
			<td>Male Wistar rats</td>
			<td>Envigo</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Size 10 surgical scalpel blades</td>
			<td>Miltex</td>
			<td>SKU: 4-110</td>
			<td></td>
		</tr>
	</tbody>
</table>

activity-based training on a treadmill with spinal cord injured wistar rats

脊髓损伤 (sci) 导致持久的缺陷, 包括流动性和多种与自主相关的功能障碍。跑步机上的 locomotor 培训 (lt) 被广泛用作 sci 人群的康复工具, 对日常生活有许多好处和改善。我们利用这种基于活动的任务特定训练 (abt) 的方法在啮齿类动物 sci 后, 既阐明了这些改进背后的机制, 并加强和改进现有的临床康复方案。我们目前的目标是确定 abt 诱导的 sci 大鼠在中度至重度挫伤后尿、肠和性功能改善的机制。在固定每个单独的动物在一个定制的可调背心, 他们被固定在一个多功能的体重支持机制, 降低到一个修改的三线跑步机, 并协助在步法训练 58分钟, 每天一次, 为期10周。这种设置允许训练四足动物和前肢动物, 以及两个不同的未训练组。有体重支持的受过四倍训练的动物由在场的技术人员协助, 协助在必要时进行适当的后肢放置, 而前肢训练的动物则在尾端饲养, 以确保与跑步机, 没有负重。一个未受过训练的 sci 群动物被放置在一个马具中, 并在跑步机旁边休息, 而另一个控制 sci 组则留在附近训练室的家笼中。这种模式允许同时训练多个 sci 动物, 从而使其更省时, 此外还确保我们的临床前动物模型尽可能接近临床表现, 特别是在身体方面重量支持与手动协助。

按照这一培训协议, 有文献表明, 只有 qt 动物表现出优越的运动功能相比, 其他组 18.然而, 由于我们实验室的性质, 我们的主要重点是调查基于活动的任务特定训练 (abt) 的非运动优势, 包括膀胱、肠道和性功能。例如, 我们以前公布的数据显示, lt 导致 sci 大鼠 qt 和 ft 组多尿运动诱导减少 (图 4)17。此外, 在 qt 和 ft 组中, 没有发现损伤引起的肾脏生长因子β (tgf-β) 转化表达的减少, 表明免疫反应改变, 它们的 tgf-β水平与假 (无损伤) 动物相似。在同一项研究17中, 在安乐死和组织采集之前进行清醒膀胱测量。假组、qt 组和 ft 组的膀胱收缩最大振幅在假体、qt 和 ft 组之间没有显著差异, 而 nt 组仍有明显变化。这些数据共同表明, 肾脏健康和膀胱功能的运动结果是积极的, 从而改善了 sci 后的泌尿系统功能。
sci 人群中多尿的机制目前尚不清楚, 但很可能是多因素 32.例如, 有些人假设, 当 sci 个人坐在轮椅上时, 下肢的液体汇集会导致液体超载, 并在姿势移位 (如从坐着到躺着)时增加液体的消除。这样的解释并不成立的临床前模型, 这导致我们最初专注于精氨酸加压素 (avp), 控制体内液体稳态的激素, 可以通过运动进行调节。avp 通过激活肾脏中的 v2 受体来控制液体的稳态, 这有助于从肾脏收集管道中吸收水分 34.从试点实验 (慢性时间点与一个病变严重程度-210 kdyn 冲击力) 的初步证据表明运动 (lt 和 ft) 对 v2 受体水平在大鼠肾脏的有益的影响 (图 5)。
<img alt="Figure 1" class="xfigimg" src="/files/ftp_upload/58983/58983fig1.jpg"> 图 1: 为雄性 wistar 大鼠量身定制的线束.qt 和 nt 动物都被放置在相同类型的夹克 (a) 中, 允许在 qt 动物的情况下使用后肢。有额外的皮带缝在英国 ft 动物 (b) 的马具上, 以提高后肢, 确保没有体重支持。马具的大型钩环材料部分允许轻松地调整不同大小的动物, 并随着时间的推移对单个动物的大小进行任何变化。<a href="https://www.jove.com/files/ftp_upload/58983/58983fig1large.jpg" target="_blank">请点击这里查看此图的较大版本.</a>
<img alt="Figure 2" class="xfigimg" src="/files/ftp_upload/58983/58983fig2.jpg"> 图 2: 培训站设置.在跑步机周围的体重支撑机制, 包括 nt (最左边)、qt (中) 或 ft (右) 组。<a href="https://www.jove.com/files/ftp_upload/58983/58983fig2large.jpg" target="_blank">请点击这里查看此图的较大版本.</a>
<img alt="Figure 3" class="xfigimg" src="/files/ftp_upload/58983/58983fig3.jpg"> 图 3: 与动物的训练站.顶部 (a) 和 (b) 侧视图, 显示对线束的体重支持机制和附件支撑夹的位置。请注意, ft 动物 (b) 的后肢是在跑步机带上抬起和脱落的。插入物 (c) 描绘了固定在线束上的夹子的更近视图。<a href="https://www.jove.com/files/ftp_upload/58983/58983fig3large.jpg" target="_blank">请点击这里查看此图的较大版本.</a>
<img alt="Figure 4" class="xfigimg" src="/files/ftp_upload/58983/58983fig4.jpg"> 图4:abt 对 sci 后大鼠多尿的影响.在 sci (*; p < 0.05) 之后, 尿量总量 (a) 增加, 在 qt 和 ft 组进行了9周的 lt 训练后, 恢复到接近基线的水平, 但 nt 组相对于受过训练的组仍有增加 (#; p < 0.05)。与基线相比, 所有组的尿量均在9周时增加, 空隙体积增加 (b)。必须指出, 所有群体的空洞数量 (c) 和水量 (d) 保持不变。值是指±标准误差。此数字在作者允许的情况下重新公布17。<a href="https://www.jove.com/files/ftp_upload/58983/58983fig4large.jpg" target="_blank">请点击这里查看此图的较大版本.</a>
<img alt="Figure 5" class="xfigimg" src="/files/ftp_upload/58983/58983fig5.jpg"> 图 5: abt 对大鼠肾脏的影响.4组大鼠5组 (共 20只) 的大鼠肾 v2 受体水平的西方印迹结果, 显示了a组和组平均密度测量分析结果 (使用 imagej;od = b 组中的光学密度), 表明 sci 后慢性时间点 (12周) 的受体显著减少 (*; p < 0.05), 每天接受 10周 abt 的群体的受体相对于基线 (假手术控制) 没有减少。错误条表示标准错误。<a href="https://www.jove.com/files/ftp_upload/58983/58983fig5large.jpg" target="_blank">请点击这里查看此图的较大版本.</a>
<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always"><tbody>
<tr>
<td colspan="3">凯塔明-西拉津剂量图</td>
			<td></td>
			<td></td>
		</tr>
<tr>
<td>有效剂量:</td>
			<td colspan="4">使用 100 mgml 氯胺酮库存和 20 mg/xylazine 库存 * * *</td>
		</tr>
<tr>
<td>80克/千克氯胺酮</td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
<tr>
<td>10 mg/kg xylazine</td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
<tr>
<td colspan="5">1.0 ml 混合物注射 = 0.62 ml 氯胺酮库存 (100 mg/ml) + 0.62 ml xylazine 库存 (20 mg/ml)</td>
		</tr>
<tr>
<td>动物重量</td>
			<td colspan="2">混合注射</td>
			<td>动物重量</td>
			<td>混合注射</td>
		</tr>
<tr>
<td>(g)</td>
			<td>(毫升)</td>
			<td></td>
			<td>(g)</td>
			<td>(毫升)</td>
		</tr>
<tr>
<td>100元</td>
			<td>0.13</td>
			<td></td>
			<td>275</td>
			<td>0.36</td>
		</tr>
<tr>
<td>105</td>
			<td>0.14</td>
			<td></td>
			<td>285</td>
			<td>0.37</td>
		</tr>
<tr>
<td>1110</td>
			<td>0.14</td>
			<td></td>
			<td>290</td>
			<td>0.38</td>
		</tr>
<tr>
<td>115</td>
			<td>0.15</td>
			<td></td>
			<td>300元</td>
			<td>0.39</td>
		</tr>
<tr>
<td>1120</td>
			<td>0.16</td>
			<td></td>
			<td>305</td>
			<td>0。4</td>
		</tr>
<tr>
<td>125</td>
			<td>0.16</td>
			<td></td>
			<td>310</td>
			<td>0。4</td>
		</tr>
<tr>
<td>1130</td>
			<td>0.17</td>
			<td></td>
			<td>315</td>
			<td>0.41</td>
		</tr>
<tr>
<td>135</td>
			<td>0.18</td>
			<td></td>
			<td>320</td>
			<td>0.42</td>
		</tr>
<tr>
<td>140</td>
			<td>0.18</td>
			<td></td>
			<td>325</td>
			<td>0.42</td>
		</tr>
<tr>
<td>145</td>
			<td>0.19</td>
			<td></td>
			<td>330</td>
			<td>0.43</td>
		</tr>
<tr>
<td>150人</td>
			<td>0。2</td>
			<td></td>
			<td>335</td>
			<td>0.44</td>
		</tr>
<tr>
<td>155</td>
			<td>0。2</td>
			<td></td>
			<td>340</td>
			<td>0.44</td>
		</tr>
<tr>
<td>160</td>
			<td>0.21</td>
			<td></td>
			<td>345</td>
			<td>0.45</td>
		</tr>
<tr>
<td>165</td>
			<td>0.21</td>
			<td></td>
			<td>350</td>
			<td>0.46</td>
		</tr>
<tr>
<td>170</td>
			<td>0.22</td>
			<td></td>
			<td>355</td>
			<td>0.46</td>
		</tr>
<tr>
<td>175</td>
			<td>0.23</td>
			<td></td>
			<td>360</td>
			<td>0.47</td>
		</tr>
<tr>
<td>180元</td>
			<td>0.23</td>
			<td></td>
			<td>365</td>
			<td>0.47</td>
		</tr>
<tr>
<td>185</td>
			<td>0.24</td>
			<td></td>
			<td>370</td>
			<td>0.48</td>
		</tr>
<tr>
<td>190</td>
			<td>0.25</td>
			<td></td>
			<td>375</td>
			<td>0.49</td>
		</tr>
<tr>
<td>195</td>
			<td>0.25</td>
			<td></td>
			<td>380</td>
			<td>0.49</td>
		</tr>
<tr>
<td>200人</td>
			<td>0.26</td>
			<td></td>
			<td>385</td>
			<td>0。5</td>
		</tr>
<tr>
<td>205</td>
			<td>0.27</td>
			<td></td>
			<td>390</td>
			<td>0.51</td>
		</tr>
<tr>
<td>210</td>
			<td>0.27</td>
			<td></td>
			<td>395</td>
			<td>0.51</td>
		</tr>
<tr>
<td>215</td>
			<td>0.28</td>
			<td></td>
			<td>400人</td>
			<td>0.52</td>
		</tr>
<tr>
<td>220</td>
			<td>0.29</td>
			<td></td>
			<td>410</td>
			<td>0.53</td>
		</tr>
<tr>
<td>225</td>
			<td>0.29</td>
			<td></td>
			<td>420</td>
			<td>0.55</td>
		</tr>
<tr>
<td>230</td>
			<td>0。3</td>
			<td></td>
			<td>430</td>
			<td>0.56</td>
		</tr>
<tr>
<td>235</td>
			<td>0.31</td>
			<td></td>
			<td>440</td>
			<td>0.57</td>
		</tr>
<tr>
<td>240</td>
			<td>0.31</td>
			<td></td>
			<td>450元</td>
			<td>0.59</td>
		</tr>
<tr>
<td>245</td>
			<td>0.32</td>
			<td></td>
			<td>460</td>
			<td>0。6</td>
		</tr>
<tr>
<td>250人</td>
			<td>0.33</td>
			<td></td>
			<td>470</td>
			<td>0.61</td>
		</tr>
<tr>
<td>255</td>
			<td>0.33</td>
			<td></td>
			<td>480</td>
			<td>0.62</td>
		</tr>
<tr>
<td>260</td>
			<td>0.34</td>
			<td></td>
			<td>490</td>
			<td>0.64</td>
		</tr>
<tr>
<td>265</td>
			<td>0.34</td>
			<td></td>
			<td>500元</td>
			<td>0.65</td>
		</tr>
<tr>
<td>1270</td>
			<td>0.35</td>
			<td></td>
			<td>510</td>
			<td>0.66</td>
		</tr>
</tbody></table>表 1: 根据单个动物的体重绘制的麻醉剂量图。
<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always"><tbody>
<tr>
<td>培训时间 (分钟)</td>
			<td>速度 (平方米)</td>
			<td>持续时间 (分钟)</td>
		</tr>
<tr>
<td>0-1</td>
			<td>6</td>
			<td>1</td>
		</tr>
<tr>
<td>1-2</td>
			<td>8。8</td>
			<td>1</td>
		</tr>
<tr>
<td>2-3</td>
			<td>10。8</td>
			<td>1</td>
		</tr>
<tr>
<td>3-8</td>
			<td>13。2</td>
			<td>5</td>
		</tr>
<tr>
<td>8-13</td>
			<td>10。8</td>
			<td>5</td>
		</tr>
<tr>
<td>13-28</td>
			<td>13。2</td>
			<td>15</td>
		</tr>
<tr>
<td>28-33</td>
			<td>10。8</td>
			<td>5</td>
		</tr>
<tr>
<td>33-38</td>
			<td>6</td>
			<td>5</td>
		</tr>
<tr>
<td>38-43</td>
			<td>8。8</td>
			<td>5</td>
		</tr>
<tr>
<td>43-58</td>
			<td>13。2</td>
			<td>15</td>
		</tr>
</tbody></table>表 2: 跑步机速度设置的训练方案应与每次速度所花费的时间相对应。

Watch this Scientific Journal Video about Activity-based Training on a Treadmill with Spinal Cord Injured Wistar Rats at JoVE.com

Activity-based Training on a Treadmill with Spinal Cord Injured Wistar Rats

该方案展示了我们的基于活动的运动跑步机训练大鼠脊髓损伤 (sci) 模型。其中包括四足组和前肢组, 以及两种不同类型的未训练控制组。调查人员能够使用该协议评估对 sci 大鼠的训练效果。

脊髓损伤 wistar 大鼠跑步机的作业训练

Spinal cord injury (SCI) results in lasting deficits that include both mobility and a multitude of autonomic-related dysfunctions. Locomotor training (LT) ...

运动和运动训练通常用于脊髓损伤患者人群的康复治疗。基于活动的培训的主要优点是，许多功能优势已被记录，在临床前动物研究，并在临床实践中。演示这个程序的将是詹姆斯·阿姆斯特朗和克里斯汀·亚贝里，他们都是高级研究技术人员，还有我实验室的研究生杰森·甘贝尔。在脊髓损伤前的一周内，处理每只大鼠五到十分钟，每天一次，为期五天。受伤当天，在受伤地点剃掉第一只200至225克大鼠的背部的头发，并在低处将麻醉动物放在加热垫上。管理眼部润滑剂、长效普通抗生素、镇痛剂和五毫升生理盐水。根据椎骨张胆估计目标病变水平的位置，并使用数字10手术刀在动物的胸椎上，直接在中线椎骨上方，进行大约5厘米的切口。使用海莫斯特，收回侧肌，使用4-0丝缝合线。对于中胸挫伤，使用荣格去除覆盖的T7椎骨层，暴露脊髓的T8-T9水平，并使用挫伤装置进行挫伤。分娩后，使用4-0直径的单丝缝合在一起的肌肉和筋膜在脊髓上。用九毫米手术伤口夹合合皮肤。再注射五毫升的生理盐水。然后将动物放在单独的清洁笼子里，放在加热垫上，在24-48小时内进行密切监测。使用手动信任

activity-based-training-on-treadmill-with-spinal-cord-injured-wistar

Research

JoVE Journal

Neuroscience

7.9K 次观看.  University of Louisville. 运动和运动训练通常用于脊髓损伤患者人群的康复治疗。基于活动的培训的主要优点是，许多功能优势已被记录，在临床前动物研究，并在临床实践中。演示这个程序的将是詹姆斯·阿姆斯特朗和克里斯汀·亚贝里，他们都是高级研究技术人员，还有我实验室的研究生杰森·甘贝尔。在脊髓损伤前的一周内，处理每只大鼠五到十分钟，每天一次，为期五天。受伤当天，在受伤?...

视频: Activity-based Training on a Treadmill with Spinal Cord Injured Wistar Rats

Spinal Cord Transection in the Larval Zebrafish

Mammals fail in sensory and motor recovery following spinal cord injury due to lack of axonal regrowth below the level of injury as well as an inability to reinitiate spinal neurogenesis. However, some anamniotes including the zebrafish Danio rerio exhibit both sensory and functional recovery even after complete transection of the spinal cord. The adult zebrafish is an established model organism for studying regeneration following spinal cord injury, with sensory and motor recovery by 6&#160;weeks post-injury. To take advantage of in vivo analysis of the regenerative process available in the transparent larval zebrafish as well as genetic tools not accessible in the adult, we use the larval zebrafish to study regeneration after spinal cord transection. Here we demonstrate a method for reproducibly and verifiably transecting the larval spinal cord. After transection, our data shows sensory recovery beginning at 2&#160;days post-injury (dpi), with the C-bend movement detectable by 3 dpi and resumption of free swimming by 5 dpi. Thus we propose the larval zebrafish as a companion tool to the adult zebrafish for the study of recovery after spinal cord injury.

After spinal transection, adult zebrafish have functional recovery by six weeks post-injury. To take advantage of larval transparency and faster recovery, we present a method for transecting the larval spinal cord. After transection, we observe sensory recovery beginning at 2&#160;days post-injury, and C-bend movement by 3 days post-injury.

在斑马鱼幼虫脊髓横断

Mammals fail in sensory and motor recovery following spinal cord injury due to lack of axonal regrowth below the level of injury as well as an inability ...

科研

神经科学

A Procedure for Implanting a Spinal Chamber for Longitudinal In Vivo Imaging of the Mouse Spinal Cord

Studies in the mammalian neocortex have enabled unprecedented resolution of cortical structure, activity, and response to neurodegenerative insults by repeated, time-lapse in vivo imaging in live rodents. These studies were made possible by straightforward surgical procedures, which enabled optical access for a prolonged period of time without repeat surgical procedures. In contrast, analogous studies of the spinal cord have been previously limited to only a few imaging sessions, each of which required an invasive surgery. As previously described, we have developed a spinal chamber that enables continuous optical access for upwards of 8 weeks, preserves mechanical stability of the spinal column, is easily stabilized externally during imaging, and requires only a single surgery. Here, the design of the spinal chamber with its associated surgical implements is reviewed and the surgical procedure is demonstrated in detail. Briefly, this video will demonstrate the preparation of the surgical area and mouse for surgery, exposure of the spinal vertebra and appropriate tissue debridement, the delivery of the implant and vertebral clamping, the completion of the chamber, the removal of the delivery system, sealing of the skin, and finally, post-operative care. The procedure for chronic in vivo imaging using nonlinear microscopy will also be demonstrated. Finally, outcomes, limitations, typical variability, and a guide for troubleshooting are discussed.

A Procedure for Implanting a Spinal Chamber for Longitudinal In Vivo Imaging of the Mouse Spinal Cord

In this video, we describe a procedure for implanting a chronic optical imaging chamber over the dorsal spinal cord of a live mouse. The chamber, surgical procedure, and chronic imaging are reviewed and demonstrated.

一个程序，植入脊柱商会纵<em&gt;在体内</em&gt;鼠标脊髓成像

Studies in the mammalian neocortex have enabled unprecedented resolution of cortical structure, activity, and response to neurodegenerative insults by ...

Experimental Strategies to Bridge Large Tissue Gaps in the Injured Spinal Cord after Acute and Chronic Lesion

After a spinal cord injury (SCI) a scar forms in the lesion core which hinders axonal regeneration. Bridging the site of injury after an insult to the spinal cord, tumor resections, or tissue defects resulting from traumatic accidents can aid in facilitating general tissue repair as well as regenerative growth of nerve fibers into and beyond the affected area. Two experimental treatment strategies are presented: (1) implantation of a novel microconnector device into an acutely and completely transected thoracic rat spinal cord to readapt severed spinal cord tissue stumps, and (2) polyethylene glycol filling of the SCI site in chronically lesioned rats after scar resection. The chronic spinal cord lesion in this model is a complete spinal cord transection which was inflicted 5 weeks before treatment. Both methods have recently achieved very promising outcomes and promoted axonal regrowth, beneficial cellular invasion and functional improvements in rodent models of spinal cord injury.
The mechanical microconnector system (mMS) is a multi-channel system composed of polymethylmethacrylate (PMMA) with an outlet tubing system to apply negative pressure to the mMS lumen thus pulling the spinal cord stumps into the honeycomb-structured holes. After its implantation into the 1 mm tissue gap the tissue is sucked into the device. Furthermore, the inner walls of the mMS are microstructured for better tissue adhesion.
In the case of the chronic spinal cord injury approach, spinal cord tissue - including the scar-filled lesion area - is resected over an area of 4 mm in length. After the microsurgical scar resection the resulting cavity is filled with polyethylene glycol (PEG 600) which was found to provide an excellent substratum for cellular invasion, revascularization, axonal regeneration and even compact remyelination in vivo.

Severe spinal cord injuries often result in tissue defects. Two possibilities are described to successfully bridge such gaps to promote tissue adaptation, regenerative responses and functional improvement in rats via implantation of a mechanical microconnector system after acute injury and five weeks after complete spinal cord transection.

实验策略急，慢性损伤后桥的损伤脊髓组织大差距

After a spinal cord injury (SCI) a scar forms in the lesion core which hinders axonal regeneration. Bridging the site of injury after an insult to the ...

Laminectomy and Spinal Cord Window Implantation in the Mouse

This protocol describes a method for spinal cord laminectomy and glass window implantation for in vivo imaging of the mouse spinal cord. An integrated digital vaporizer is utilized to achieve a stable plane of anesthesia at a low-flow rate of isoflurane. A single vertebral spine is removed, and a commercially available cover-glass is overlaid on a thin agarose bed. A 3D-printed plastic backplate is then affixed to the adjacent vertebral spines using tissue adhesive and dental cement. A stabilization platform is used to reduce motion artifact from respiration and heartbeat. This rapid and clamp-free method is well-suited for acute multi-photon fluorescence microscopy. Representative data are included for an application of this technique to two-photon microscopy of the spinal cord vasculature in transgenic mice expressing eGFP:Claudin-5 &#8212; a tight junction protein.

This protocol describes implantation of a glass window onto the spinal cord of a mouse to facilitate visualization by intravital microscopy.

拉米内切手术和脊髓窗口植入小鼠

This protocol describes a method for spinal cord laminectomy and glass window implantation for in vivo imaging of the mouse spinal cord. An integrated ...

Isolation of Adult Spinal Cord Nuclei for Massively Parallel Single-nucleus RNA Sequencing

Probing an individual cell&#39;s gene expression enables the identification of cell type and cell state. Single-cell RNA sequencing has emerged as a powerful tool for studying transcriptional profiles of cells, particularly in heterogeneous tissues such as the central nervous system. However, dissociation methods required for single cell sequencing can lead to experimental changes in the gene expression and cell death. Furthermore, these methods are generally restricted to fresh tissue, thus limiting studies on archival and bio-bank material. Single nucleus RNA sequencing (snRNA-Seq) is an appealing alternative for transcriptional studies, given that it accurately identifies cell types, permits the study of tissue that is frozen or difficult to dissociate, and reduces dissociation-induced transcription. Here, we present a high-throughput protocol for rapid isolation of nuclei for downstream snRNA-Seq. This method enables isolation of nuclei from fresh or frozen spinal cord samples and can be combined with two massively parallel droplet encapsulation platforms.

Here, we present a protocol to rapidly isolate high-quality nuclei from the fresh or frozen tissue for downstream massively parallel RNA sequencing. We include detergent-mechanical and hypotonic-mechanical tissue disruption and cell lysis options, both of which can be used for isolation of nuclei.

成人脊髓细胞核在大规模平行单核 RNA 测序中的分离

Probing an individual cell&#39;s gene expression enables the identification of cell type and cell state. Single-cell RNA sequencing has emerged as a ...

Intra-Arterial Delivery of Neural Stem Cells to the Rat and Mouse Brain: Application to Cerebral Ischemia

Neural stem cell (NSC) therapy is an emerging innovative treatment for stroke, traumatic brain injury and neurodegenerative disorders. As compared to intracranial delivery, intra-arterial administration of NSCs is less invasive and produces a more diffuse distribution of NSCs within the brain parenchyma. Further, intra-arterial delivery allows the first-pass effect in the brain circulation, lessening the potential for trapping of cells in peripheral organs, such as liver and spleen, a complication associated with peripheral injections. Here, we detail the methodology, in both mice and rats, for delivery of NSCs through the common carotid artery (mouse) or external carotid artery (rat) to the ipsilateral hemisphere after an ischemic stroke. Using GFP-labeled NSCs, we illustrate the widespread distribution achieved throughout the rodent ipsilateral hemisphere at 1 d, 1 week and 4 weeks after postischemic delivery, with a higher density in or near the ischemic injury site. In addition to long-term survival, we show evidence of differentiation of GFP-labeled cells at 4 weeks. The intra-arterial delivery approach described here for NSCs can also be used for administration of therapeutic compounds, and thus has broad applicability to varied CNS injury and disease models across multiple species.

A method for delivering neural stem cells, adaptable for injecting solutions or suspensions, through the common carotid artery (mouse) or external carotid artery (rat) after ischemic stroke is reported. Injected cells are distributed broadly throughout the brain parenchyma and can be detected up to 30 d after delivery.

神经干细胞向大鼠和小鼠大脑的动脉内传递：对脑缺血的应用

Neural stem cell (NSC) therapy is an emerging innovative treatment for stroke, traumatic brain injury and neurodegenerative disorders. As compared to ...

A Minimally Invasive, Fast Spinal Cord Lateral Hemisection Technique for Modeling Open Spinal Cord Injuries in Rats

Open spinal cord injury techniques modeling laceration-like injuries are time-consuming and invasive because they involve laminectomy. This new technique eliminates laminectomy by removing two spinous processes and lifting, then tilting the caudal vertebral arch. The surgical area opens up without the need for laminectomy. Lateral hemisection is then performed with direct visible control under a microscope. The trauma is minimized, requiring only a small bone wound. 
This technique has several advantages: it is faster and, therefore, less of a burden for the animal, and the bone wound is smaller. Because the laminectomy is eliminated, there is less chance for unwanted injury to the spinal cord, and there are no bone splinters that can cause problems (bone splinters embedded in the spinal cord can cause swelling and secondary damage). The vertebral canal remains intact. The main limitation is that the hemisection can only be performed in the intervertebral spaces. 
The results show that this technique can be performed much faster than the traditional surgical approach, using laminectomy (11 min vs. 35 min). This technique can be useful for researchers working with animal models of open spinal cord injury as it is widely adaptable and does not require any additional specialized instrumentation.

Here, we describe a new, fast technique modeling open spinal cord injury in rats that eliminates laminectomy. Lateral hemisection is performed while viewing through a microscope. The technique is versatile and can also be used in the cervical, thoracic, and lumbar regions of the spinal cord of other animals.

一种微创、快速脊髓侧向半切技术，用于模拟大鼠开放性脊髓损伤

Open spinal cord injury techniques modeling laceration-like injuries are time-consuming and invasive because they involve laminectomy. This new technique ...

Establishing a Mouse Contusion Spinal Cord Injury Model Based on a Minimally Invasive Technique

Using minimally invasive methods to model spinal cord injury (SCI) can minimize behavioral and histological differences between experimental animals, thereby improving the reproducibility of the experiments.
These methods need two requirements to be fulfilled: clarity of the surgical anatomical pathway and simplicity and convenience of the laboratory device. Crucially for the operator, a clear anatomical pathway provides minimally invasive exposure, which avoids additional damage to the experimental animal during the surgical procedures and allows the animal to maintain a consistent and stable anatomical morphology during the experiment.
In this study, the use of a novel integrated platform called the SCI coaxial platform for spinal cord injury in small animals to expose the T9 level spinal cord in a minimally invasive way and stabilize and immobilize the vertebra of mice using a vertebral stabilizer is researched, and, finally, a coaxial gravity impactor is used to contuse the spinal cord of mice to approach different degrees of T9 spinal cord injury. Finally, histological results are provided as a reference for the readers.

Minimally invasive techniques and a simple laboratory device improve the reproducibility of the spinal cord injury model by reducing operative damage to the experimental animals and allowing anatomical morphology maintenance. The method is worthwhile because the reliable results and reproducible procedure facilitate investigations of the mechanisms of disease reparation.

基于微创技术的小鼠挫伤脊髓损伤模型的建立

Using minimally invasive methods to model spinal cord injury (SCI) can minimize behavioral and histological differences between experimental animals, ...

Electromagnetic Controlled Closed-Head Model of Mild Traumatic Brain Injury in Mice

Highly reproducible animal models of traumatic brain injury (TBI), with well-defined pathologies, are needed for testing therapeutic interventions and understanding the mechanisms of how a TBI alters brain function. The availability of multiple animal models of TBI is necessary to model the different aspects and severities of TBI seen in people. This manuscript describes the use of a midline closed head injury (CHI) to develop a mouse model of mild TBI. The model is considered mild because it does not produce structural brain lesions based on neuroimaging or gross neuronal loss. However, a single impact creates enough pathology that cognitive impairment is measurable at least 1 month after injury. A step-by-step protocol to induce a CHI in mice using a stereotaxically guided electromagnetic impactor is defined in the paper. The benefits of the mild midline CHI model include the reproducibility of the injury-induced changes with low mortality. The model has been temporally characterized up to 1 year after the injury for neuroimaging, neurochemical, neuropathological, and behavioral changes. The model is complementary to open skull models of controlled cortical impact using the same impactor device. Thus, labs can model both mild diffuse TBI and focal moderate-to-severe TBI with the same impactor.

The protocol describes mild traumatic brain injury in a mouse model. In particular, a step-by-step protocol to induce a mild midline closed head injury and the characterization of the animal model is fully explained.

小鼠轻度创伤性脑损伤的电磁控制闭头模型

Highly reproducible animal models of traumatic brain injury (TBI), with well-defined pathologies, are needed for testing therapeutic interventions and ...

小鼠脊髓中小胶质细胞的提取和酶促纯化

Rapid and Efficient Enrichment of Mouse Spinal Cord Microglia

The vertebral column defines a vertebrate and shapes the spinal canal, a cavity that encloses and safeguards the spinal cord. Proper development and function of the mammalian central nervous system rely significantly on the activity of resident macrophages known as microglia. Microglia display heterogeneity and multifunctionality, enabling distinct gene expression and behavior within the spinal cord and brain. Numerous studies have explored cerebral microglia function, detailing purification methods extensively. However, the purification of microglia from the spinal cord in mice lacks a comprehensive description. In contrast, the utilization of a highly purified collagenase, as opposed to an unrefined extract, lacks reporting within central nervous system tissues. In this study, the vertebral column and spinal cord were excised from 8-10 week-old C57BL/6 mice. Subsequent digestion employed a highly purified collagenase, and microglia purification utilized a density gradient. Cells underwent staining for flow cytometry, assessing viability and purity through CD11b and CD45 staining. Results yielded an average viability of 80% and a mean purity of 95%. In conclusion, manipulation of mouse microglia involved digestion with a highly purified collagenase, followed by a density gradient. This approach effectively produced substantial spinal cord microglia populations.

作者聚焦：小胶质细胞对脊髓异质性和纯化的研究 

Microglia are regarded as some of the most versatile cells in the body, capable of morphological and functional adaptation. Their heterogeneity and multifunctionality enable the maintenance of brain homeostasis, while also being linked to various neurological pathologies. Here, a technique for purifying spinal cord microglia is described.

小鼠脊髓小胶质细胞的快速高效富集

The vertebral column defines a vertebrate and shapes the spinal canal, a cavity that encloses and safeguards the spinal cord. Proper development and ...