Name: development of an algorithm to perform a comprehensive study of autonomic dysreflexia in animals with high spinal cord injury using a telemetry device
Uploaded: 2016-07-29T13:00:00.000+00:00
Duration: 6 min 51 s
Description: Watch this Scientific Journal Video about Development of an Algorithm to Perform a Comprehensive Study of Autonomic Dysreflexia in Animals with High Spinal Cord Injury Using a Telemetry Device at JoVE

这项研究如果资助的健康研究的加拿大研究所和心脏及中风BC和育空地区的基础。我们要感谢Rayshad Gopaul先生和谢利McErlane博士在动物保健技术支持和专业知识。

雄性Wistar（HSD:WI的Wistar）大鼠7周龄，体重300-350克本实验中使用。所有大鼠维持12小时的光/暗周期，并获得标准的实验室大鼠饲料和水随意。所有实验程序符合由加拿大议会关于动物饲养建立先导，以护理和使用实验动物，并授予伦理委员会批准由英国哥伦比亚大学。手术和动物护理是根据我们实验室的标准程序（拉姆齐等，2010）17进行。 1.动物的制备:外科手术<ol><li> 遥测装置植入 注:协议被实施根据制造商说明和我们以前的与设备的注入经验:16 <ol><li>与恩诺沙星预治疗的动物，一旦一个连续3天每天（10毫克/千克，S.C。）手术前。 </li><li>在手术前从笼中取出固体食物9-13小时。在此期间可能会提供软食营养代餐。 </li><li>麻醉用4％异氟烷在感应腔中诱导剂量和使用贝恩电路约1.5％异氟烷（1-1.5升/分钟氧气）的维持剂量的动物。用脚趾捏反射和角膜反射来评估动物是否是在麻醉手术平面。呼吸必须监测以评估是否麻醉深度太高（ 即 ，非常缓慢呼吸速率和喘息，苍白粘膜）或过低（ 即，快速，浅呼吸指示动物可能不会在麻醉的手术平面）。在手术期间和麻醉大约每5分钟的外科平面的重复监测调整异氟烷剂量是必要的。 注:所有手术使用的无菌技术进行:具体而言，医生穿外科擦洗，帽，口罩和无菌手套。开配戴手套过程观察，高压灭菌含有手术器械使用无菌技术被打开手术组和动物开始手术之前叠加。手术工具是尖灭菌烫珠手术间（最多5次手术）。外科海绵和纱布使用前高压灭菌。在手术过程中，将动物保持在维持在37℃的循环水毯。 </li><li>辖丁丙诺啡（0.02毫克/千克，SC）立即手术前。此外，管理酮洛芬（5毫克/千克; SC）手术前，以提供多峰镇痛。辖布比卡因（5毫克/公斤，SC）立即手术前，以提供本地线路块。在剃须腹部，应用交替氯己定和70％的乙醇拭子（各三次），接着碘伏为至少1分钟，并用70％的乙醇（具有新开醇每次擦拭）除去。皮肤的擦拭在同心的方式制成在切口线的中心开始并向外盘旋（以便不通过"回/来回"运动污染的区域）。用无菌纱布拭子和各个应用程序之间丢弃。此过程开始动物手术前重复三次。虽然修剪和手术擦洗，保持对异氟醚下约1.5％维持剂量连接到贝恩非再呼吸电路的鼻锥动物。以维持所需要的动物在麻醉的手术计划调整异氟烷水平。 </li><li>做一个5厘米长的中线腹部切口，通过肌肉和腹膜露出肠子切割。 </li><li>缩回肠（这将暴露在后腹壁降主动脉），盖上无菌手术纱布肠道用2-3的林格溶液毫升润湿并保持湿润与视需要，以防止组织脱水通过过程的其他林格。 </li><li>钝解剖围绕主动脉和主动脉从利用虹膜铲刀的下腔静脉分离。 </li><li>地方结扎（8-0丝缝合线）围绕主动脉的最外膜层，只是远端肾动脉的喙和尾部端。 </li><li>解除结扎简要地阻塞血液流动，并在1-2毫米前的水平与一个25号针头弯曲在位于头90°髂分叉刺穿主动脉。 </li><li>使用20号弯曲针引导遥测装置的导管的尖端进入主动脉。推进吻侧，以确保该针尖仅仅是远端肾动脉<ol><li>利用组织粘合剂少量修复它。去除结扎并确保有由于导管或组织粘合剂无血流闭塞。 </li></ol></li><li>固定遥测装置的主体成使用4-0丝线缝合腹壁。使用4-0薇乔和4-0普理灵缝线关闭MUSC乐和皮肤，分别。 </li></ol></li><li> 遥测植入手术后护理 注意:对于前14天手术后的动物应每天至少3次检查中，除了一个详细监测评估每天一次。评估动物在7类:体重，体貌（疼痛是基于迹象，如增加眼睛，鼻子和前爪周围看到卟啉生产和面部鬼脸体征评估），行为/活动，排便，皮肤（包括中恒，立毛，切口愈合不良及皮下积液），水化和呼吸。 <ol><li>喂动物，水果，谷物品种，软食营养代餐，并提供水自由采食，遥测装置植入后6天。动物应该有扣缴第一天操作后定期固体啮齿动物饲料。注入动物温热乳酸林格溶液（5毫升）皮下第3天手术后，经外观脱水的症状。管理在早上和晚上检查（皮肤帐篷苍白耳/粘膜的迹象）林格溶液（5毫升）在必要时对动物进行评估。 </li><li>辖恩诺沙星（10毫克/千克，SC）和酮洛芬（5毫克/公斤，SC），每天一次，连续3天手术后。手术后的以下3天早晚（ 即，每12小时）期间管理丁丙诺啡（0.02毫克/公斤，SC）。如果疼痛/窘迫的迹象都指出丁丙诺啡中旬的一天剂量（0.02毫克/千克，SC）可施用。 </li></ol></li><li> 脊髓全横断 注:让动物从遥测植入手术T3完全离断术前恢复至少14天。 <ol><li>预对待动物的预防性恩诺沙星（10毫克/公斤，SC）为SCI手术前三天。 </li><li>如前所述麻醉4％异氟烷的诱导剂量下的动物。进一步，用注射盐酸氯胺酮（70毫克/千克，IP）和盐酸右美托咪（0.5毫克/千克，IP）的动物。 注意:对于完全离断手术，造成异氟醚血管扩张往往呈现止血的问题，一旦线切断。可以通过使用氯胺酮/右美托咪鸡尾酒来防止这个问题。动物应麻醉20-25分钟，并使用脊髓横断手术以下阿替美唑（1毫克/千克，SC）逆转。 </li><li>手术前施用皮下丁丙诺啡（0.02毫克/千克），和酮洛芬（5毫克/千克）。辖布比卡因（5毫克/公斤，SC）立即手术前，以提供本地线路块。在剃须腹部，交替氯己定和70％乙醇的刷子（各三次），接着碘伏应适用于至少1分钟，并用70％的乙醇（具有新开醇每次擦拭）中除去。刷子在同心的方式作出开始在切口线的中心，并且向外盘旋（以便不通过"回/来回"运动污染的区域）。用无菌纱布和各个应用程序之间丢弃。开始动物手术前重复该过程三次。虽然修剪和手术擦洗，保持动物的鼻锥下异氟醚的约1.5％，维持剂量（按要求调整，以保持麻醉手术平面）。在整个手术动物的持续时间被保持在保持在37℃的循环水毯。 </li><li>做一个背中线切口，2厘米长，在浅肌层覆C8-T3椎体。 </li><li>布朗特以揭示硬脑膜层解剖。 </li><li>刺穿与microscissors或用25号针头的尖端硬脑膜层。 </li><li>打开在T2-T3椎间隙硬脑膜和使用microscissors执行完全横断。打开microscissors但不足够的横向破裂椎arterieS和静脉。在迅速的运动横切脊髓和搜索在动物后肢夸张的抽搐。确认通过喙和尾部脊髓树桩目视间隔完全横断。 </li><li>确认后，将树桩之间明胶海绵，以达到止血。施加足够的明胶海绵止血和脊髓的树桩分开。 </li><li>使用4-0薇乔和4-0普理灵缝线关闭肌肉和皮肤，分别为。脊髓损伤手术后，给动物预温热乳酸林格氏溶液（5毫升，SC 30℃），并允许它们在温度受控的环境中恢复。 </li></ol></li><li> 脊髓横断术后护理 注意:对于前14天手术后的动物应每天3次检查中，除了每天一次一个详细的监测评估。 14天后详细的监测评估可能隔日重复。评估动物在7 catego里斯:体重，体貌（疼痛是基于迹象，如卟啉和面部鬼脸评估），行为/活动，排便，皮肤（包括中恒，竖毛，切口愈合不良及皮下积液），水化和呼吸。用于第一5-7天气囊应该检查并表示每天至少3次。 <ol><li>喂动物，水果，谷物品种，营养软食代餐，并提供水自由采食，脊髓横断手术后8-14天。注入用温热乳酸林格氏溶液（5毫升）皮下注射动物手术后的第3天，在脱水症状。管理的检查过程中林格溶液（5毫升）在必要时对动物进行评估。 注:脊髓横断后，大鼠表现出后肢瘫痪，并采用屈曲和前肢，这是在这个损伤模型保存扩展走动。老鼠缺乏躯干和尾巴支持。为了方便它们在笼子里的运动，橡胶格应置于下滚燕麦床上用品。这使得大鼠与前爪抓住网格，并在笼子容易移动到获取食物和水。床上用品应每天更换。在脊髓横断后第7天，燕麦床上用品应该由它们原来的木片床上用品所取代。因为给大鼠丁丙诺啡燕麦卷被褥使用可以开发PICA在他们吃的床上用品，这可能会导致食管impactions。燕麦与燕麦食管吸湿嵌塞尚未观察到。 </li><li>辖恩诺沙星（10毫克/千克，SC）和酮洛芬（5毫克/公斤，SC）每天一次连续3天手术后。手术后的以下3天早晚（ 即 ，每12小时）期间管理丁丙诺啡（0.02毫克/公斤，SC）。如果疼痛/窘迫的迹象都指出丁丙诺啡中旬的一天剂量可以。 </li></ol></li></ol> 2.遥测血流参数监测<ol><li>收集每天打逐拍动脉血压（ABP）和核心体温24小时，预先和后SCI。采样动脉血压和核心体温在使用遥测装置24小时的块1000赫兹。 </li><li>放置的SmartPad下动物笼换能器的无线再充电，并用于接收来自换能器的数字遥测信号。 注意:输出电压随后被传递到连接到计算机，该组织从每个通道的数据流的配置。该电压被转换为连续的动脉血压和温度记录。可视利用数据采集软件的采样数据，如LabChart。 </li><li>丢弃动物监测过程中收集的遥测数据，以及在10分钟之前立即监测和监测10分钟后。 </li></ol> 3.自发INCID评估自主神经异常反射的分配办法（AD） 注:频率，严重程度，和自发的广告事件的持续时间是用我们自己的小说AD检测 JAVA平台软件（ 图4）开发了一种算法评估。新算法已经发展到自动根据之前和SCI利用图2指定的参数后24小时SBP和HR遥测录音自发的广告事件。 <ol><li>在提取使用采集软件的计息期限从原材料遥测数据SBP和HR值。 注意:采集软件检测并确定从连续ABP录音SBP峰的值。心脏速率是通过确定相邻收缩压峰之间的时间距离外推。 </li><li>上传遥测录音在A列interbeat区间，在B列SBP值，列C MAP值和时间的CSV文件我N个列D（图4A）。 </li><li>使用软件（图4B-D），指定相关生理HR范围（即 R到R的间隔）180 BPM和625 BPM之间的SBP录音。指定下心率和最小 心率最大面板的软件中的相关人力资源范围。 </li><li>通过240秒的移动平均窗口中创建了SBP和HR的阈值。表明移动平均阈值在广告持续时间阈值面板上的分析窗口长度。 </li><li>在20毫米汞柱均线基线上方设置一个SBP换位门槛。表明，在移动平均阈值换位BP面板换位值。 </li><li>隔离收缩压峰簇超出换位阈用峰 - 峰间隔少于2秒和大于10秒的持续时间。指定峰使用峰间间隔BP和杜峰间隔使用峰丛间隔BP面板比间隔。 </li><li>组收缩压峰簇是彼此的120秒内。 注意:在彼此的120秒内检测到的电位的AD事件将被分组为单个事件。指定连续SBP集群之间可以区分的广告持续时间阈值面板独立的广告事件的最长持续时间。 </li><li>确认是否收缩压峰簇群通过检测在40 BPM或更大的人​​力资源的下降与潜在自发的AD事件相关联。 <ol><li>在潜在事件的发病人力资源值的平均值的10％（指定要在上心脏速率这个百分比下降平均范围面板）。从电势事件结束的心脏速率值的平均值的75％（指定要在下部这个百分比心脏率下降平均范围面板）。 </li><li>减去从较高心脏速率阈下心脏速率的阈值，以确保一个correspon40 BPM或更大的下降鼎。使用指定的心脏率下降限制面板的心脏率下降限制。 </li></ol></li><li>一旦面板已经排满，按确定 。检测出的AD事件的图形表示呈现，包括收缩压的尖峰和相关联的人力资源数据（ 图4C-D）。一个输出excel文件也与升压反应（毫米汞柱），持续时间（秒），最大值收缩压（毫米汞柱），最小的HR（BPM），每个检测出的AD事件的心率下降（BPM）中产生。 </li></ol> 4.结直肠扩张有意引导学生AD 注:AD诱发的严重程度可以通过结直肠扩张（CRD），模仿肠例行3,18,19临床相关的刺激来确定。 <ol><li>验证弗伦奇导管的囊（直的，10法语，将3ml 35厘米，游离胶乳）不被用2ml空气充气它泄漏。 </li><li>第约束通过包裹在毛巾，让血压和心率稳定了半个小时ê动物。 </li><li>放置润滑剂在导管的尖端。放置一个标记在导管用永久性标记，2厘米从气囊的中间。导管经由肛门插入结肠插入直到标记在开口刚刚可见。固定导管与外科胶带的尾部。重新放置在他的家笼中的动物包裹放在接收器匹配的SmartPad在动物体内植入的传感器。允许动物的血压正常化10分钟。 </li><li>导管的插入和相关的适应期（ 即，10分钟）后，注入2毫升的空气的气球在10秒内，使血压和心率稳定。保持扩张1分钟。确保在导管的尖端保持在直肠内。重要的是不要过度膨胀球囊（ 即，将 3ml +），以避免对结肠的破裂。 注意:腹胀为1分钟的持续时间与＃160;允许升压反应的发展和与AD的感应情节相关联的压力感受器介导的HR降适当的时间。此外，此1分钟内还提供了足够的时间，心血管指数的正常化。 </li><li>重复扩张每次​​试验的3倍，试验间10分钟的最小间隔每日重复试验。一旦评估完成，从动物取出导尿管。 </li><li>平均节拍由拍数据超过1秒的间隔和报告SBP在多个试验最高涨幅和最大跌幅人力资源。 </li></ol>

<ol>
	<li>Krassioukov, A., Claydon, V. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+clinical+problems+in+cardiovascular+control+following+spinal+cord+injury:+an+overview.">The clinical problems in cardiovascular control following spinal cord injury: an overview.</a> Progress in brain research. 152, 223-229 (2006).</li><li>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=Autonomic+function+following+cervical+spinal+cord+injury.">Autonomic function following cervical spinal cord injury.</a> Respiratory physiology &amp; neurobiology. 169, 157-164 (2009).</li><li>Teasell, R. W., Arnold, J. M. O., Krassioukov, A., Delaney, G. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cardiovascular+consequences+of+loss+of+supraspinal+control+of+the+sympathetic+nervous+system+after+spinal+cord+injury.">Cardiovascular consequences of loss of supraspinal control of the sympathetic nervous system after spinal cord injury.</a> Archives of physical medicine and rehabilitation. 81, 506-516 (2000).</li><li>Alan, N., 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=Recurrent+autonomic+dysreflexia+exacerbates+vascular+dysfunction+after+spinal+cord+injury.">Recurrent autonomic dysreflexia exacerbates vascular dysfunction after spinal cord injury.</a> The Spine Journal. 10, 1108-1117 (2010).</li><li>Eltorai, I., Kim, R., Vulpe, M., Kasravi, H., Ho, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Fatal+cerebral+hemorrhage+due+to+autonomic+dysreflexia+in+a+tetraplegic+patient:+case+report+and+review.">Fatal cerebral hemorrhage due to autonomic dysreflexia in a tetraplegic patient: case report and review.</a> Spinal Cord. 30, 355-360 (1992).</li><li>Pan, 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=Intracerebral+hemorrhage+secondary+to+autonomic+dysreflexia+in+a+young+person+with+incomplete+C8+tetraplegia:+A+case+report.">Intracerebral hemorrhage secondary to autonomic dysreflexia in a young person with incomplete C8 tetraplegia: A case report.</a> Archives of physical medicine and rehabilitation. 86, 591-593 (2005).</li><li>Ho, C., 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=Autonomic+dysreflexia+and+myocardial+ischemia.">Autonomic dysreflexia and myocardial ischemia.</a> Spinal cord. 48, 714-715 (2010).</li><li>Wan, D., Krassioukov, A. V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Life-threatening+outcomes+associated+with+autonomic+dysreflexia:+A+clinical+review.">Life-threatening outcomes associated with autonomic dysreflexia: A clinical review.</a> The journal of spinal cord medicine. 37, 2-10 (2014).</li><li>Mathias, C. J., Frankel, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cardiovascular+control+in+spinal+man.">Cardiovascular control in spinal man.</a> Annual review of physiology. 50, 577-592 (1988).</li><li>Hubli, M., Krassioukov, A. V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ambulatory+Blood+Pressure+Monitoring+in+Spinal+Cord+Injury:+Clinical+Practicability.">Ambulatory Blood Pressure Monitoring in Spinal Cord Injury: Clinical Practicability.</a> Journal of neurotrauma. 31, 789-797 (2014).</li><li>Liu, N., Fougere, R., Zhou, M., Nigro, M., 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=Autonomic+dysreflexia+severity+during+urodynamics+and+cystoscopy+in+individuals+with+spinal+cord+injury.">Autonomic dysreflexia severity during urodynamics and cystoscopy in individuals with spinal cord injury.</a> Spinal cord. 51, 863-867 (2013).</li><li>Phillips, A. A., Elliott, S. L., Zheng, M. M., Krassioukov, A. V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Selective+alpha+adrenergic+antagonist+reduces+severity+of+transient+hypertension+during+sexual+stimulation+after+spinal+cord+injury.">Selective alpha adrenergic antagonist reduces severity of transient hypertension during sexual stimulation after spinal cord injury.</a> Journal of neurotrauma. , (2014).</li><li>Maiorov, D. N., Fehlings, M. G., Weaver, L. C., Krassioukov, A. V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Relationship+between+severity+of+spinal+cord+injury+and+abnormalities+in+neurogenic+cardiovascular+control+in+conscious+rats.">Relationship between severity of spinal cord injury and abnormalities in neurogenic cardiovascular control in conscious rats.</a> Journal of neurotrauma. 15, 365-374 (1998).</li><li>Maiorov, D. N., Weaver, L. C., Krassioukov, A. V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Relationship+between+sympathetic+activity+and+arterial+pressure+in+conscious+spinal+rats.">Relationship between sympathetic activity and arterial pressure in conscious spinal rats.</a> American Journal of Physiology-Heart and Circulatory Physiology. 41, H625 (1997).</li><li>Rabchevsky, A. 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=Effects+of+gabapentin+on+muscle+spasticity+and+both+induced+as+well+as+spontaneous+autonomic+dysreflexia+after+complete+spinal+cord+injury.">Effects of gabapentin on muscle spasticity and both induced as well as spontaneous autonomic dysreflexia after complete spinal cord injury.</a> Frontiers in physiology. 3, (2012).</li><li>Mayorov, D. N., Adams, M. A., Krassioukov, A. V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Telemetric+blood+pressure+monitoring+in+conscious+rats+before+and+after+compression+injury+of+spinal+cord.">Telemetric blood pressure monitoring in conscious rats before and after compression injury of spinal cord.</a> Journal of neurotrauma. 18, 727-736 (2001).</li><li>Ramsey, J. B., 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=Care+of+rats+with+complete+high-thoracic+spinal+cord+injury.">Care of rats with complete high-thoracic spinal cord injury.</a> Journal of neurotrauma. 27, 1709-1722 (2010).</li><li>Krassioukov, A. V., Furlan, J. C., Fehlings, M. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Autonomic+dysreflexia+in+acute+spinal+cord+injury:+an+under-recognized+clinical+entity.">Autonomic dysreflexia in acute spinal cord injury: an under-recognized clinical entity.</a> Journal of neurotrauma. 20, 707-716 (2003).</li><li>Krogh, K., Mosdal, C., Laurberg, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Gastrointestinal+and+segmental+colonic+transit+times+in+patients+with+acute+and+chronic+spinal+cord+lesions.">Gastrointestinal and segmental colonic transit times in patients with acute and chronic spinal cord lesions.</a> Spinal cord. 38, 615-621 (2000).</li><li>Krassioukov, A. V., Johns, D. G., Schramm, L. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sensitivity+of+sympathetically+correlated+spinal+interneurons,+renal+sympathetic+nerve+activity,+and+arterial+pressure+to+somatic+and+visceral+stimuli+after+chronic+spinal+injury.">Sensitivity of sympathetically correlated spinal interneurons, renal sympathetic nerve activity, and arterial pressure to somatic and visceral stimuli after chronic spinal injury.</a> Journal of neurotrauma. 19, 1521-1529 (2002).</li><li>Braga, V. A., Prabhakar, N. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Refinement+of+telemetry+for+measuring+blood+pressure+in+conscious+rats.">Refinement of telemetry for measuring blood pressure in conscious rats.</a> Journal of the American Association for Laboratory Animal Science: JAALAS. 48, 268 (2009).</li><li>Whitesall, S. E., Hoff, J. B., Vollmer, A. P., D'Alecy, L. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Comparison+of+simultaneous+measurement+of+mouse+systolic+arterial+blood+pressure+by+radiotelemetry+and+tail-cuff+methods.">Comparison of simultaneous measurement of mouse systolic arterial blood pressure by radiotelemetry and tail-cuff methods.</a> American Journal of Physiology-Heart and Circulatory Physiology. 286, H2408-H2415 (2004).</li></ol>

The authors have nothing to disclose.

该协议描述了详细的实施这将与一个遥测装置相结合，在SCI-动物（ 图1B）ABP的长期深入分析的新的JAVA平台的AD检测软件。这是第一个软件，它允许为ABP图案表征，因为它们偶尔发生在一天的持续时间，以检测自发广告事件。充分表征T3 SCI动物模型可以说明检测自发AD的频率，升压反应和持续的软件功能的能力。随着检测自发广告事件，该软件可以分析ABP振荡和辨别血流动力学参数的?...

自主神经异常反射（AD）是在颈部或高胸节急性或慢性脊髓损伤（SCI）后的个体威胁生命的紧急情况和通常的特征在于持续性高血压和心动过缓1情节。 AD主要是由脊髓下行通道，通常要控制交感神经活性和血管张力1-4脊髓交感神经节前神经元提供了从脊髓上输入中心的破坏引起的。高达300毫米汞柱的AD发作的特征在于收缩压（SBP）尖峰并且如果不治疗会导致癫痫发作，颅内出血，心肌梗塞，甚至死亡5-8。各种有害和非伤害性刺激的作为AD的触发，包括肠和膀胱膨胀，痉挛，压疮，膀胱导尿或医源性程序9-12。 AD的RESP的时间发展ONSE以SCI已经在人类9和动物模型13,14了研究。通常这些研究已经使用了"诱导AD"的方法（ 即尿流动力学，在人类或动物的结直肠扩张阴茎vibrostimulations）确定AD的时间发展。这种方法由于需要在分离的时间点重复评估可能排除AD的时间发展的一个精确确定的限制。在人类中使用的24小时的血压监测器在预定的时间间隔进行串行血压测量。这种技术最近已用于监测慢性SCI自发产生的AD。在动物模型中，固态压力传感器正在越来越多地用于长期监控打逐击败动脉血压。近日，Rabchesvky 等 。 （2012），开发出了提取平均动脉压的二分之一的平均值的算法（MAP）的一个对一个移动平均阈值15相比Ð。基于MAP的峰是10毫米汞柱或更大的阈值以上的同时用10 BPM或更大的HR一滴自发公元事件进行了表征。 在这里，有一个建于公元检测算法一种新型的JAVA软件呈现。这个算法通过检测动脉血压（ABP）和心脏速率（HR），该指示一个自然发生​​的AD事件的预先确定的图案。用户能够手动调整所有输入变量的软件，使得&#39;检测算法"可以容易地定制，以所关注的具体参数。该软件还能够二分法ABP和HR成一个给定的时期，使得血流动力学参数的昼夜节律可以分析16。在本手稿，详细说明中给出，用于植入遥测设备和进行SCI手术手术技术。前还提供了相对于所述的AD检测软件的后处理能力和功能如何心血管amples被改变后的SCI。为了比较的目的，从被称为结直肠扩张诱导的AD的方法得到的方法和结果（CRD）也示出。

<table><tbody><tr><td>11 Male Wistar Rats -Hsd-WI (250-300g)</td><td>Envigo (formerly Harlan Laboratories)&amp;nbsp;</td><td>141</td><td></td></tr><tr><td>Lab Chart (PowerLab&amp;reg; Data Acquisition System)</td><td>AD Instruments&amp;nbsp;</td><td></td><td></td></tr><tr><td>Pressure Telemeter&amp;nbsp;</td><td>Millar Inc.</td><td>RP-TRM54P</td><td></td></tr><tr><td>Configurator</td><td>Millar Inc.</td><td>TR190</td><td></td></tr><tr><td>SmartPad</td><td>Millar Inc.</td><td>TR180</td><td></td></tr><tr><td>Isoflurane (Aerrane)</td><td>Baxter Corp.</td><td>DIN: 02225875</td><td></td></tr><tr><td>Enrofloxacin (Baytril)</td><td>Bayer Healthcare</td><td>DIN: 02169428</td><td></td></tr><tr><td>5-0 Silk Sutures</td><td>Ethicon</td><td>S182</td><td></td></tr><tr><td>4-0 Vicryl Subcuticular</td><td>Ethicon</td><td>J496G</td><td></td></tr><tr><td>Buprenorphine (Temgesic)</td><td>Reckitt Benckiser</td><td>DIN: 0281250</td><td></td></tr><tr><td>Bupivicaine Hydrochloride (Marcaine 0.5%)</td><td>Hospira Healthcare Corp.&amp;nbsp;</td><td>DIN: 02305909</td><td></td></tr><tr><td>Ketoprofen (Anafen)</td><td>Merial</td><td>DIN: 02150999</td><td></td></tr><tr><td>Ketamine Hydrochloride (Vetalar)</td><td>Bioniche</td><td>DIN: 01989529</td><td></td></tr><tr><td>Dexmedetomidine Hydrochloride (Domitor)</td><td>Pfizer</td><td>DIN: 02333929</td><td></td></tr><tr><td>Lactated Ringer&#039;s Solution&amp;nbsp;</td><td>Braun Medical Inc.</td><td>DIN: 01931636</td><td></td></tr><tr><td>Gelfoam #12</td><td>Pharmacia &amp;amp; Upjohn Company</td><td>03603-14-1</td><td></td></tr><tr><td>Microscissors</td><td>Fine Science Tools</td><td>15003-008</td><td></td></tr><tr><td>Iris Spatulae</td><td>Fine Science Tools</td><td>10094-13</td><td></td></tr><tr><td>10 French 35&amp;nbsp;cm Foley Catheter</td><td>Coloplast&amp;nbsp;</td><td>AA6110</td><td></td></tr><tr><td>Dietgel&amp;reg;&amp;nbsp;</td><td>Clear H2O, Westbrook, ME</td><td>76A</td><td></td></tr><tr><td>LabDiet Rodent Diet 5001</td><td>Purina Mills (PMI&amp;reg;)</td><td>5001</td><td></td></tr><tr><td>Chlorhexadine (Hibitane)</td><td>Wyeth Animal Health, Guelph, Ontario</td><td>DIN 00245097</td><td></td></tr><tr><td>Atipamezole Hydrochloride(Antisedan)</td><td>Orion Pharma</td><td>DIN: 02237744</td><td></td></tr></tbody></table>

development of an algorithm to perform a comprehensive study of autonomic dysreflexia in animals with high spinal cord injury using a telemetry device

Spinal cord injury (SCI) is a debilitating neurological condition characterized by somatic and autonomic dysfunctions. In particular, SCI above the mid-thoracic level can lead to a potentially life-threatening hypertensive condition called autonomic dysreflexia (AD) that is often triggered by noxious or non-noxious somatic or visceral stimuli below the level of injury. One of the most common triggers of AD is the distension of pelvic viscera, such as during bladder and bowel distension or evacuation. This protocol presents a novel pattern recognition algorithm developed for a JAVA platform software to study the fluctuations of cardiovascular parameters as well as the number, severity and duration of spontaneously occurring AD events. The software is able to apply a pattern recognition algorithm on hemodynamic data such as systolic blood pressure (SBP) and heart rate (HR) extracted from telemetry recordings of conscious and unrestrained animals before and after thoracic (T3) complete transection. With this software, hemodynamic parameters and episodes of AD are able to be detected and analyzed with minimal experimenter bias.

使用遥测技术，动脉血压是在1000赫兹的频率连续24小时取样。使用LabChart动脉血压（ABP）的说明性记录示于图1B。样品ABP被插入降主动脉固态压力传感器监测。小说JAVA平台AD检测软件能够提取相关收缩压（毫米汞柱）峰（ 图1C）。我们还可以提取来自相邻SBP峰（ 图1D）之间的时间间隔的心率（BPM）。 SBP和HR通过在1000赫兹采样节拍ABP从节拍...

Watch this Scientific Journal Video about Development of an Algorithm to Perform a Comprehensive Study of Autonomic Dysreflexia in Animals with High Spinal Cord Injury Using a Telemetry Device at JoVE

Development of an Algorithm to Perform a Comprehensive Study of Autonomic Dysreflexia in Animals with High Spinal Cord Injury Using a Telemetry Device

The catheter of a telemetry device is implanted into the abdominal aorta in order to continuously collect beat-by-beat hemodynamic data from animals pre and post-high thoracic spinal cord transection. A novel JAVA software was employed to analyze hemodynamic parameters as well as frequency and intensity of spontaneous episodes of autonomic dysreflexia.

一个算法的开发与高位脊髓损伤使用遥测装置进行自主神经异常反射的动物的综合研究

Spinal cord injury (SCI) is a debilitating neurological condition characterized by somatic and autonomic dysfunctions. In particular, SCI above the ...

development-an-algorithm-to-perform-comprehensive-study-autonomic

Research

JoVE Journal

Medicine

7.8K 次观看.  Faculty of Medicine, University of British Columbia. The catheter of a telemetry device is implanted into the abdominal aorta in order to continuously collect beat-by-beat hemodynamic data from animals pre and post-high thoracic spinal cord transection. A novel JAVA software was employed to analyze hemodynamic parameters as well as frequency and intensity of spontaneous episodes of autonomic dysreflexia.

视频: Development of an Algorithm to Perform a Comprehensive Study of Autonomic Dysreflexia in Animals with High Spinal Cord Injury Using a Telemetry Device

Acute and Chronic Tactile Sensory Testing after Spinal Cord Injury in Rats

Spinal cord injury (SCI) impairs sensory systems causing allodynia1-8. To identify cellular and molecular causes of allodynia, sensitive and valid sensory testing in rat SCI models is needed. However, until recently, no single testing approach had been validated for SCI so that standardized methods have not been implemented across labs. Additionally, available testing methods could not be implemented acutely or when severe motor impairments existed, preventing studies of the development of SCI-induced allodynia3. Here we present two validated sensory testing methods using von Frey Hair (VFH) monofilaments which quantify changes in tactile sensory thresholds after SCI4-5. One test is the well-established Up-Down test which demonstrates high sensitivity and specificity across different SCI severities when tested chronically5. The other test is a newly-developed dorsal VFH test that can be applied acutely after SCI when allodynia develops, prior to motor recovery4-5. Each VFH monofilament applies a calibrated force when touched to the skin of the hind paw until it bends. In the up-down method, alternating VFHs of higher or lower forces are used on the plantar L5 dermatome to delineate flexor withdrawal thresholds. Successively higher forces are applied until withdrawal occurs then lower force VFHs are used until withdrawal ceases. The tactile threshold reflects the force required to elicit withdrawal in 50% of the stimuli. For the new test, each VFH is applied to the dorsal L5 dermatome of the paw while the rat is supported by the examiner. The VFH stimulation occurs in ascending order of force until at least 2 of 3 applications at a given force produces paw withdrawal. Tactile sensory threshold is the lowest force to elicit withdrawal 66% of the time. Acclimation, testing and scoring procedures are described. Aberrant trials that require a retest and typical trials are defined. Animal use was approved by Ohio State University Animal Care and Use Committee.

We describe two tactile sensory testing methods for acute or chronic periods of spinal cord injury in rats. These validated procedures can detect the development and maintenance of allodynia-like sensations.

在大鼠脊髓损伤后的急性和慢性触觉测试

Spinal cord injury (SCI) impairs sensory systems causing allodynia1-8. To identify cellular and molecular causes of allodynia, sensitive and valid sensory ...

科研

医学

Using Laser Doppler Imaging and Monitoring to Analyze Spinal Cord Microcirculation in Rat

Laser Doppler flowmetry (LDF) is a noninvasive method for blood flow (BF) measurement, which makes it preferable for measuring microcirculatory alterations of the spinal cord. In this article, our goal was to use both Laser Doppler imaging and monitoring to analyze the change of BF after spinal cord injury. Both the laser Doppler image scanner and the probe/monitor were being employed to obtain each readout. The data of LDPI provided a local distribution of BF, which gave an overview of perfusion around the injury site and made it accessible for comparative analysis of BF among different locations. By intensely measuring the probing area over a period of time, a combined probe was used to simultaneously measure the BF and oxygen saturation of the spinal cord, showing overall spinal cord perfusion and oxygen supply. LDF itself has a few limitations, such as relative flux, sensitivity to movement, and biological zero signal. However, the technology has been applied in clinical and experimental study due to its simple setup and rapid measurement of BF.

Here we present a combination of laser Doppler perfusion imaging (LDPI) and laser Doppler perfusion monitoring (LDPM) to measure spinal cord local blood flows and oxygen saturation (SO2), as well as a standardized procedure for introducing spinal cord trauma on rat.

激光多普勒成像与监测在大鼠脊髓微循环分析中的应用

Laser Doppler flowmetry (LDF) is a noninvasive method for blood flow (BF) measurement, which makes it preferable for measuring microcirculatory ...

行为学

Use of a Wireless Video-EEG System to Monitor Epileptiform Discharges Following Lateral Fluid-Percussion Induced Traumatic Brain Injury

The lateral fluid percussion injury (FPI) model is well established and has been used to study TBI and post-traumatic epilepsy (PTE). However, considerable variability has been reported for the specific parameters used in different studies that have employed this model, making it difficult to harmonize and interpret the results between laboratories. For example, variability has been reported regarding the size and location of the craniectomy, how the Luer lock hub is placed relative to the craniectomy, the atmospheric pressure applied to the dura and the duration of the pressure pulse. Each of these parameters can impact injury severity, which directly correlates with the incidence of PTE. This has been manifested as a wide range of mortality rates, righting reflex times and incidence of convulsive seizures reported. Here we provide a detailed protocol for the method we have used to help facilitate harmonization between studies. We used FPI in combination with a wireless EEG telemetry system to continuously monitor for electrographic changes and detect seizure activity.&#160; FPI is induced by creating a 5 mm craniectomy over the left hemisphere, between the Bregma and Lambda and adjacent to the lateral ridge. A Luer lock hub is secured onto the skull over the craniectomy. This hub is connected to the FPI device, and a 20-millisecond pressure pulse is delivered directly to the intact dura through pressure tubing connected to the hub via a twist lock connector. Following recovery, rats are re-anesthetized to remove the hub. Five 0.5 mm, stainless steel EEG electrode screws are placed in contact with the dura through the skull and serve as four recording electrodes and one reference electrode. The electrode wires are collected into a pedestal connector which is secured into place with bone cement. Continuous video/EEG recordings are collected for up to 4 weeks post TBI.

Here we present a protocol to induce severe TBI with the lateral fluid percussion injury (FPI) model in adult, male Wistar rats. We also demonstrate the use of a wireless telemetry system to collect continuous video-EEG recordings and monitor for epileptiform discharges consistent with post-traumatic epileptogenesis.&#160;

使用无线视频-EEG系统监测侧流冲击后癫痫的放电诱发创伤性脑损伤

The lateral fluid percussion injury (FPI) model is well established and has been used to study TBI and post-traumatic epilepsy (PTE). However, ...

A Radio-telemetric System to Monitor Cardiovascular Function in Rats with Spinal Cord Transection and Embryonic Neural Stem Cell Grafts

High thoracic or cervical spinal cord injury (SCI) can lead to cardiovascular dysfunction. To monitor cardiovascular parameters, we implanted a catheter connected to a radio transmitter into the femoral artery of rats that underwent a T4 spinal cord transection with or without grafting of embryonic brainstem-derived neural stem cells expressing green fluorescent protein. Compared to other methods such as cannula insertion or tail-cuff, telemetry is advantageous to continuously monitor blood pressure and heart rate in freely moving animals. It is also capable of long term multiple data acquisitions. In spinal cord injured rats, basal cardiovascular data under unrestrained condition and autonomic dysreflexia in response to colorectal distension were successfully recorded. In addition, cardiovascular parameters before and after SCI can be compared in the same rat if a transmitter is implanted before a spinal cord transection. One limitation of the described telemetry procedure is that implantation in the femoral artery may influence the blood supply to the ipsilateral hindlimb.

We present a protocol for using a radio-telemetric system to record cardiovascular parameters in T4 spinal cord transected rats eight weeks after embryonic brainstem neural stem cell grafting into the lesion site. Telemetry is an advanced technique to accurately evaluate cardiovascular function in conscious freely moving spinal cord injured rats.

无线电遥测系统，以监测大鼠脊髓横断和胚胎神经干细胞移植在心血管功能

High thoracic or cervical spinal cord injury (SCI) can lead to cardiovascular dysfunction. To monitor cardiovascular parameters, we implanted a catheter ...

Novel Approach for Simultaneous Recording of Renal Sympathetic Nerve Activity and Blood Pressure with Intravenous Infusion in Conscious, Unrestrained Mice.

Renal sympathetic nerves contribute significantly to both physiological and pathophysiological phenomena. Evaluating renal sympathetic nerve activity (RSNA) is of great interest in many areas of research such as chronic kidney disease, hypertension, heart failure, diabetes and obesity. Unequivocal assessment of the role of the sympathetic nervous system is thus imperative for proper interpretation of experimental results and understanding of disease processes. RSNA has been traditionally measured in anesthetized rodents, including mice. However, mice usually exhibit very low systemic blood pressure and hemodynamic instability for several hours during anesthesia and surgery. Meaningful interpretation of RSNA is confounded by this non-physiological state, given the intimate relationship between sympathetic nervous tone and cardiovascular status. To address this limitation of traditional approaches, we developed a new method for measuring RSNA in conscious, freely-moving mice. Mice were chronically instrumented with radio-telemeters for continuous monitoring of blood pressure as well as a jugular venous infusion catheter and custom-designed bipolar electrode for direct recording of RSNA. Following a 48-72 hour recovery period, survival rate was 100% and all mice behaved normally. At this time-point, RSNA was successfully recorded in 80% of mice, with viable signals acquired up to 4 and 5 days post-surgery in 70% and 50% of mice, respectively. Physiological blood pressures were recorded in all mice (116&plusmn;2 mmHg; n=10). Recorded RSNA increased with eating and grooming, as well-established in the literature. Furthermore, RSNA was validated by ganglionic blockade and modulation of blood pressure with pharmacological agents. Herein, an effective and manageable method for clear recording of RSNA in conscious, freely-moving mice is described.

Anesthetized mice exhibit non-physiological systemic blood pressure, which precludes meaningful assessment of autonomic tone given the intimate relationship between blood pressure and the autonomic nervous system. Thus, a novel method to simultaneously record renal sympathetic nerve activity and blood pressure with intravenous infusion in conscious mice is outlined.

清醒、无拘无束的小鼠静脉输液同时记录肾交感神经活动和血压的新方法。

Renal sympathetic nerves contribute significantly to both physiological and pathophysiological phenomena. Evaluating renal sympathetic nerve activity ...

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

Spinal cord injury (SCI) results in lasting deficits that include both mobility and a multitude of autonomic-related dysfunctions. Locomotor training (LT) on a treadmill is widely used as a rehabilitation tool in the SCI population with many benefits and improvements to daily life. We utilize this method of activity-based task-specific training (ABT) in rodents after SCI to both elucidate the mechanisms behind such improvements and to enhance and improve upon existing clinical rehabilitation protocols. Our current goal is to determine the mechanisms underlying ABT-induced improvements in urinary, bowel, and sexual function in SCI rats after a moderate to severe level of contusion. After securing each individual animal in a custom-made adjustable vest, they are secured to a versatile body weight support mechanism, lowered to a modified three-lane treadmill and assisted in step-training for 58 minutes, once a day for 10 weeks. This setup allows for the training of both quadrupedal and forelimb-only animals, alongside two different non-trained groups. Quadrupedal-trained animals with body weight support are aided by a technician present to assist in stepping with proper hind limb placement as necessary, while forelimb-only trained animals are raised at the caudal end to ensure no hind limb contact with the treadmill and no weight-bearing. One non-trained SCI group of animals is placed in a harness and rests next to the treadmill, while the other control SCI group remains in its home cage in the training room nearby. This paradigm allows for the training of multiple SCI animals at once, thus making it more time-efficient in addition to ensuring that our pre-clinical animal model mimics the clinical representation as close as possible, particularly with respect to the body weight support with manual assistance.

This protocol demonstrates our model of activity-based locomotor treadmill training for rats with spinal cord injury (SCI). Included is both quadrupedal and forelimb-only groups, in addition to two distinct types of non-trained control groups. Investigators are able to assess training effects on SCI rats using this protocol.

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) ...

神经科学

Thoracic Spinal Cord Hemisection Surgery and Open-Field Locomotor Assessment in the Rat

Spinal cord injury (SCI) causes disturbances in motor, sensory, and autonomic function below the level of the lesion. Experimental animal models are valuable tools to understand the neural mechanisms involved in locomotor recovery after SCI and to design therapies for clinical populations. There are several experimental SCI models including contusion, compression, and transection injuries that are used in a wide variety of species. A hemisection involves the unilateral transection of the spinal cord and disrupts all ascending and descending tracts on one side only. Spinal hemisection produces a highly selective and reproducible injury in comparison to contusion or compression techniques that is useful for investigating neural plasticity in spared and damaged pathways associated with functional recovery. We present a detailed step-by-step protocol for performing a thoracic hemisection at the T8 vertebral level in the rat that results in an initial paralysis of the hindlimb on the side of the lesion with graded spontaneous recovery of locomotor function over several weeks. We also provide a locomotor scoring protocol to assess functional recovery in the open-field. The locomotor assessment provides a linear recovery profile and can be performed both early and repeatedly after injury in order to accurately screen animals for appropriate time points in which to conduct more specialized behavioral testing. The hemisection technique presented can be readily adapted to other transection models and species, and the locomotor assessment can be used in a variety of SCI and other injury models to score locomotor function.

The rat thoracic spinal hemisection is a valuable and reproducible model of unilateral spinal cord injury to investigate the neural mechanisms of locomotor recovery and treatment efficacy. This article includes a detailed step-by-step guide to perform the hemisection procedure and to assess locomotor performance in an open-field arena.

胸脊髓截断手术和大鼠开放场运动评估

Spinal cord injury (SCI) causes disturbances in motor, sensory, and autonomic function below the level of the lesion. Experimental animal models are ...

In Vivo Intracellular Recording of Type-Identified Rat Spinal Motoneurons During Trans-Spinal Direct Current Stimulation

Intracellular recording of spinal motoneurons in vivo provides a &#8220;gold standard&#8221; for determining the cells&#8217; electrophysiological characteristics in the intact spinal network and holds significant advantages relative to classical in vitro or extracellular recording techniques. An advantage of in vivo intracellular recordings is that this method can be performed on adult animals with a fully mature nervous system, and therefore many observed physiological mechanisms can be translated to practical applications. In this methodological paper, we describe this procedure combined with externally applied constant current stimulation, which mimics polarization processes occurring within spinal neuronal networks. Trans-spinal direct current stimulation (tsDCS) is an innovative method increasingly used as a neuromodulatory intervention in rehabilitation after various neurological injuries as well as in sports. The influence of tsDCS on the nervous system remains poorly understood and the physiological mechanisms behind its actions are largely unknown. The application of the tsDCS simultaneously with intracellular recordings enables us to directly observe changes of motoneuron membrane properties and characteristics of rhythmic firing in response to the polarization of the spinal neuronal network, which is crucial for the understanding of tsDCS actions. Moreover, when the presented protocol includes the identification of the motoneuron with respect to an innervated muscle and its function (flexor versus extensor) as well as the physiological type (fast versus slow) it provides an opportunity to selectively investigate the influence of tsDCS on identified components of spinal circuitry, which seem to be differently affected by polarization. The presented procedure focuses on surgical preparation for intracellular recordings and stimulation with an emphasis on the steps which are necessary to achieve preparation stability and reproducibility of results. The details of the methodology of the anodal or cathodal tsDCS application are discussed while paying attention to practical and safety issues.

This protocol describes in vivo intracellular recording of rat lumbar motoneurons with simultaneous trans-spinal direct current stimulation. The method enables us to measure membrane properties and to record rhythmic firing of motoneurons before, during and after anodal or cathodal polarization of the spinal cord.

在跨脊柱直流刺激期间类型识别大鼠脊柱腺瘤的 Vivo 细胞内记录

Intracellular recording of spinal motoneurons in vivo provides a &#8220;gold standard&#8221; for determining the cells&#8217; electrophysiological ...

Automated Gait Analysis to Assess Functional Recovery in Rodents with Peripheral Nerve or Spinal Cord Contusion Injury

Peripheral and central nerve injuries are mostly studied in rodents, especially rats, given the fact that these animal models are both cost-effective and a lot of comparative data has been published in the literature. This includes a multitude of assessment methods to study functional recovery following nerve injury and repair. Besides evaluation of nerve regeneration by means of histology, electrophysiology, and other in vivo and in vitro assessment techniques, functional recovery is the most important criterion to determine the degree of neural regeneration. Automated gait analysis allows recording of a vast quantity of gait-related parameters such as Paw Print Area and Paw Swing Speed as well as measures of inter-limb coordination. Additionally, the method provides digital data of the rats&#8217; paws after neuronal damage and during nerve regeneration, adding to our understanding of how peripheral and central nervous injuries affect their locomotor behavior. Besides the predominantly used sciatic nerve injury model, other models of peripheral nerve injury such as the femoral nerve can be studied by means of this method. In addition to injuries of the peripheral nervous systems, lesions of the central nervous system, e.g., spinal cord contusion can be evaluated. Valid and reproducible data assessment is strongly dependent on meticulous adjustment of the hard- and software settings prior to data acquisition. Additionally, proper training of the experimental animals is of crucial importance. This work aims to illustrate the use of computerized automated gait analysis to assess functional recovery in different animal models of peripheral nerve injury as well as spinal cord contusion injury. It also emphasizes the method&#8217;s limitations, e.g., evaluation of nerve regeneration in rats with sciatic nerve neurotmesis due to limited functional recovery. Therefore, this protocol is thought to help researchers interested in peripheral and central nervous injuries to assess functional recovery in rodent models.

Automated gait analysis is a feasible tool to evaluate functional recovery in rodent models of peripheral nerve injury and spinal cord contusion injury. While it requires only one setup to assess locomotor function in various experimental models, meticulous hard- and soft-ware adjustment and training of the animals is highly important.

自动步态分析，评估带外周神经或脊髓挫伤的啮齿动物的功能恢复

Peripheral and central nerve injuries are mostly studied in rodents, especially rats, given the fact that these animal models are both cost-effective and ...

Implantation of Combined Telemetric ECG and Blood Pressure Transmitters to Determine Spontaneous Baroreflex Sensitivity in Conscious Mice

Blood pressure (BP) and heart rate (HR) are both controlled by the autonomic nervous system (ANS) and are closely intertwined due to reflex mechanisms. The baroreflex is a key homeostatic mechanism to counteract acute, short-term changes in arterial BP and to maintain BP in a relatively narrow physiological range. BP is sensed by baroreceptors located in the aortic arch and carotid sinus. When BP changes, signals are transmitted to the central nervous system and are then communicated to the parasympathetic and sympathetic branches of the autonomic nervous system to adjust HR. A rise in BP causes a reflex decrease in HR, a drop in BP causes a reflex increase in HR.
Baroreflex sensitivity (BRS) is the quantitative relationship between changes in arterial BP and corresponding changes in HR. Cardiovascular diseases are often associated with impaired baroreflex function. In various studies reduced BRS has been reported in e.g., heart failure, myocardial infarction, or coronary artery disease. 
Determination of BRS requires information from both BP and HR, which can be recorded simultaneously using telemetric devices. The surgical procedure is described beginning with the insertion of the pressure sensor into the left carotid artery and positioning of its tip in the aortic arch to monitor arterial pressure followed by the subcutaneous placement of the transmitter and ECG electrodes. We also describe postoperative intensive care and analgesic management. After a two-week period of post-surgery recovery long-term ECG and BP recordings are performed in conscious and unrestrained mice. Finally, we include examples of high-quality recordings and the analysis of spontaneous baroreceptor sensitivity using the sequence method.

The baroreflex is a heart-rate regulation mechanism by the autonomic nervous system in response to blood-pressure changes. We describe a surgical technique to implant telemetry transmitters for continuous and simultaneous measurement of electrocardiogram and blood pressure in mice. This can determine spontaneous baroreflex sensitivity, an important prognostic marker for cardiovascular disease.

植入遥测心电图和血压递质组合以确定意识小鼠的自发压力反射敏感性

Blood pressure (BP) and heart rate (HR) are both controlled by the autonomic nervous system (ANS) and are closely intertwined due to reflex mechanisms. ...