Name: catheterization of the carotid artery and jugular vein to perform hemodynamic measures, infusions and blood sampling in a conscious rat model
Uploaded: 2015-01-30T13:00:00.000+00:00
Duration: 9 min 5 s
Description: Watch this Scientific Journal Video about Catheterization of the Carotid Artery and Jugular Vein to Perform Hemodynamic Measures, Infusions and Blood Sampling in a Conscious Rat Model at JoVE.com

Intramural NIH program supported the development of this model. Publication support was provided by Harvard Apparatus Inc. and ADInstruments Inc. The work by the authors was done as part of US government&#8211;funded research; however, the opinions expressed are not necessarily those of the National Institutes of Health.

下面介绍的程序进行批准的临床中心的动物护理和使用委员会在美国国立卫生研究院的协议的一部分。 1.准备手术<ol><li>燃气导管消毒和器械。 </li></ol> 2.准备导管<ol><li>同花顺肝素甘油（250 IU肝素/ 1毫升的丙三醇）的导管，以确保通畅，避免泄漏。塞与金属塞子（20 GA 15毫米）至保留肝素甘油导管的末端。 </li></ol> 3.准备无菌工作站<ol><li>用70％的酒精喷工作站和仪表盘。使用无菌悬垂覆盖手术台和仪表盘和地点仪器的托盘上。 </li></ol> 4，大鼠准备外科手术<ol><li>地方大鼠分别在有机玻璃室，并暴露于麻醉剂;异氟烷（3-5％）的混合编氧气，直到昏迷。从腔室中取出大鼠和从颈部的前部和后部区域剃毛皮。涂抹润滑油膏眼睛。 </li></ol> 5.导管植入<ol><li>将麻醉的大鼠（由脚趾捏确认）到手术加热表中腹的位置;把它的鼻子到鼻锥为维持麻醉自发性呼吸（异氟烷，3.5％）。轻轻擦洗手术区3次交替使用聚维酮碘和70％的酒精。使使用外科剪刀的肩甲骨之间的0.5厘米中线皮肤切口。 </li><li>重新定位大鼠在仰卧位;轻轻抑制腿部用橡皮筋表中的每一侧;维持麻醉。放置在颈部2推出无菌纱布的4x4小幅hyperextend更好的曝光。使在使用手术刀锁骨水平颈部的中线的2厘米腹侧颈部皮肤切口权利。 </li></ol> 6.右键颈内静脉穿刺置管<ol><li>使用止血钳，直截了当地解剖右颈静脉，分离出唾液腺和淋巴组织可视化和隔离容器的一个5毫米部分。用4-0丝线缝合，放置在容器的两个头部和尾部端部的松动领带最大化容器的曝光。用微型手术剪做一个切口足够大以通过导管，管线与两个连字之间的容器中，并配合在容器周围的颅结扎。 </li><li>将静脉导管插入朝向与微解剖钩和镊子的辅助心脏的容器中，并推进该导管，直到所有的PU 3F段是在容器中。使用连字在头部和尾部端以固定导管的容器中。 </li></ol> 7.左颈动脉插管<ol><li>用止血钳，直言解剖肩胛舌骨肌纵向暴露左侧颈总动脉和我Alcalase碱性蛋白酶容器的5毫米部分。确保迷走神经（白色的），从动脉完全分离。小心不要切碎或破坏神经。 </li><li>用4-0丝线缝合，放置在容器的尾端的松散的领带，领带关闭容器的颅端和尾端放置缝合上述哈巴狗夹停止血流以下的切口。用微型手术剪，使一个切口，大到足以与两个连字之间的容器通过导管，管线。将朝心脏动脉导管与微观解剖钩钳的帮助。 </li><li>使用无锁的平滑针保持器除去牛头犬钳紧之前保持在容器内的导管的所述部分。推进导管有一对镊子的同时松开针保持器慢慢直到导管的整个聚氨酯2F段是在该容器中。配合宽松的尾部结扎导管周围和血管确保，但不能因此紧固密封吨，以堵塞，导管。 </li></ol> 8.使用直止血，隧道5cm的管皮下回耳后，并通过肩胛骨之间的切口。 Exteriorize导管通过管和取出管。 9.关闭腹切口三种不锈钢绕线夹，以及背侧切口用4-0丝线缝合固定导管外置到位。 10.手术后监测和护理<ol><li>以下导尿，终止麻醉，并恢复在卧位大鼠中与纤维素寝具一个笼子。观察大鼠在2小时的时间间隔为至少4小时，或直到表示疼痛的迹象，然后每天一次。得到酮洛芬[5毫克/千克，皮下注射（SQ）]立即张贴程序和每12小时，如果大鼠显示疼痛或困难的症状。根据我们的经验，没有老鼠都需要以下初始额外镇痛疼痛升注射。观测到的特性，将额外的剂量镇痛诱发考虑包括但不限于:姿势异常;增加/减少呼吸速率，unthrifty / ungroomed，感染/切口部位的炎症，眼分泌物，竖毛（"加标haircoat"），降低活性响应于听觉或触觉刺激。 注:大鼠恢复72小时即获得从前期导尿重"10 g将纳入研究。 </li></ol> 11.连接到血液动力系统<ol><li>以下恢复，报名参加该研究的健康大鼠（体重增加&gt; 10克）。放置大鼠的Lucite腔和麻醉如前所述。 </li><li>将无菌的悬垂在手术台上用无菌止血钳，微型镊子1毫升注射器用钝针连接。 注:该止血钳的尖端应覆盖有橡胶，以避免损坏导管。将麻醉大鼠的手术表中的腹侧的位置和地点鼻子到鼻锥维持麻醉（3-5％）具有自发呼吸。 </li><li>钳动脉并用一对橡胶封止血略低于金属插针的静脉导管。用一对镊子除去金属销在导管的端部，并附加一个1ml注射器填充有肝素盐水附有一钝针。反过来，取出止血，收回0.1毫升，以确保导管的专利。 </li><li>再夹紧导管，取出针，并为此附加到一个较长的导管（预冲洗用肝素盐水），在连接到一个旋转，并拴在鞋盒笼子的顶部。连接这些导管的换能器的数据采集和记录，取样，或输注。 注:所有导管保护，螺旋簧管足够长的时间，使动物的全套动作的笼子里。 </li><li>下面导管连接，终止麻醉在恢复与纤维素床上用品笼子里的胸骨位置动物。为了保持动脉导管通畅，人工冲洗和0.05 ml肝素盐水（50 IU / ml）的每1小时24小时这与动物的定期评估一致的线。 </li></ol> 12.采血，压力监测和药物管理局<ol><li>附加空注射器活塞连接到动脉导管，收回0.6毫升在线冲洗。附加样品注射器，取样品，并返回0.6毫升在线冲洗后0.7毫升肝素生理盐水冲洗。 </li><li>使用数据采集硬件和软件来测量和记录MAP和心脏率不断。 </li><li>自动给药系统（ADDS）连接到静脉导管。 注:将调节血管加压输注速率（高，低，或0剂量）的基础上检测到正常生理范围内维持压力在MAP。 </li></ol>

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The authors have no potential conflicts of interest.

<html>调查治疗方法的疾病的临床模型就必须准确地执行血液动力学监测，因此需要血管通路的能力。在我们的脓毒症模型，动脉导管提供了系统性压力监测和静脉插管允许的标准临床治疗中的应用。标准治疗脓毒症，包括抗生素输液治疗细菌感染和流体和升压药的输注以保持在所需范围内的动脉压力。两个导管用于采血。在此设计中，可编程输注应用与自动触发基于实时的血管加压输注速率的泵平均动脉压是预先在临床环境，需要手动滴定由医务人员和在临床前设置使使用这种治疗方式可能用于研究大量动物的。 。e_content"&gt;一些并发症模型的发展经历了早期的这些包括动脉和来自紧缩静脉导管，对PU2和PU3成分的关系，（1-2％）的部分遮挡;颈静脉导管错位成小支，（&lt;1％）;恢复（无肺动脉血栓或其他器官损伤明显）后，不明原因猝死，（1-2％）通过保持异氟醚浓度2-3.5％与2升/分钟O 2流量解析。没有观察到出血，局部感染和自身造成损伤的其他常见的手术并发症。一旦外科熟练度达到后，每个过程应该能够在不超过15分钟完成。成功的测量作为健康恢复的速率和专利导管，可以预计，在超过一次充分训练。一个附加的动物被包含在每个研究的90％，以减少数据的丢失，需要的动物的总数和帐户失败导管可以是"只有融合"或死亡，由于并发症。 在此过程中最关键的步骤是将导管的设计和布局。该材料必须是柔软的尖端，以便不以刺穿血管壁无卷曲过于容易。导管的主体具有要更硬（比前端），以便能够提前到容器充分且在适当的位置固定。 图1显示管子的直径3被连接于动脉导管和2个不同直径的管子连接为静脉导管。这些部件滑动内彼此以减少导管直径在前端并与加入，以确保在容器或心脏导管的放置被保持锚粘在一起。 一旦回收，并连接到旋转和系绳，所述MAP和CVP波形应类似于图2中所表示的数据是为了保持动脉导管patenc监测过程中Y，每小时肝素盐水冲洗（0.05毫升），应执行。连续输液保持静脉导管通畅。在线路连接和冲洗，密切一定要注意，以确保没有气泡就行了。 ≥0.1ml的气泡可导致肺，脑或其他器官的栓塞。血液动力学波形记录和在研究过程中进行分析。当在x轴压缩时， 图3示出了在1小时的过程中的变化，MAP和CVP以及变化率和血管加压输注效果。 该程序有几个潜在的局限性。在我们的研究中，导管留专利长达11天。它是未知多久导管将保持专利超过这个期限。到所述适合的导管，有一个最小的血管的大小，这大致与啮齿动物的重量相关的，需要的动物纳入本研究为大于200克。使用更小的导管直径将导致增加的电阻，以减小流动通畅。达到中心静脉压措施，需要在研究期间的心房足以帐户大小和生长精确导管放置，并且可以是一个挑战。 一旦掌握，动脉和血管导管可提供依据各种啮齿动物模型需要血液动力学监测流体或疗法，血液采样，并输注同时最小化任何疼痛和痛苦的仪器中或处理后回收的。事实上，在与股静脉的先前JOVE出版物10描述导尿结合，我们已经成功地执行了此模型用3植入导管。

大鼠的动脉和静脉导管早已用于在实验室研究1,2导管可用于监测血液动力学参数，包括收缩压，舒张压和平均动脉压（MAP），心脏速率（HR）和中心静脉压（CVP） 3,4此外，这些导管允许的标准疗法或潜在疗法输注以及血液取样，以进一步分析的疾病或治疗的作用机制。因此，具有在小动物模型的血管通路是用于研究的措施的临床表现和治疗效果的关键。 研究重大疾病的根本原因，首先建立一个模型来模拟人的条件是很重要的。气管内接种细菌的已知量的已被成功地用于重现肺炎的发病机制，一个严重肺部感染，然后发育成败血症。3，5血流动力学监测措施，并提供标准的临床治疗关键是要模拟常规支持治疗。标准的临床治疗包括输液抗生素，它有助于清除潜在的感染和体液和升压药的管理，两种治疗采用脓毒症患者维持血压。5-7血管通路，特别是当我们研究的专利导管植入尤为重要研究性治疗败血症。 多年，来管理标准的临床治疗的啮齿动物的能力通过用用于构造可植入导管和输注技术来准确地随着时间的推移递送小体积的能力所需要的特性的材料的可用性是有限的。此外，触发输注响应于连续监测的特定的血流动力学度量的能力允许以一致和可再现的立场在使用较大的样本量典型的啮齿动物的研究ARD治疗模式8,9这些技术限制在材料和精密的 ​​硬件已经克服，但同时提供常规治疗，同时监测血流动力学参数，动脉和静脉血管通路必须是可用的。 该视频详细介绍了手术技术植入颈总动脉和颈总静脉导管在麻醉的老鼠。以下一个72小时的恢复期，动物将被重新麻醉，并连接到附连到用于将植入导管连接到血液动力学监测系统中的啮齿动物壳体的系绳和旋转设置。在研究过程中，系统允许用于流体输注根据任一提供一个给定的体积在一定的输注速率或自动化系统，可提供根据所检测到保持给定的压力范围内的平均动脉压的输注速率。补液系统采用可编程注射器泵与数据采集系统的数字输出，接口和由软件监视所述MAP进行控制。系绳和旋转设置允许在研究期间大鼠可以自由流动，同时监测压力，注入液体和升压药，而无需操纵动物进行血液取样。与每个2导管（在我们的设置）与硬件可扩展至监测仪表24的动物同时监视多达12仪表化动物允许很大的灵活性，用于研究多个分层组在每个实验中。 该导管法可以是使用类似的啮齿类动物模型需要血管通路的监测，取样和治疗长达11天的研究设施是有益的。如果研究机构在考虑这个程序与啮齿类动物为维持这些动物然后developin经验和适当的支撑结构克这些技能可以提供显著节省成本，减少每个插管大鼠总成本从约1,800至不到50美元（包括天真大鼠，导管组件和技术成本），提高调度的灵活性（天真的老鼠都是现成的与公司的交付时间表植入老鼠）。这个过程的成功依赖于执行导尿个体的技能。精通此过程中所需的培训动物已经从20至40不等的大鼠与&gt; 90％导致导管通畅率。

<table><tbody><tr><td>Straight micro dissecting forceps</td><td>Roboz</td><td>RS-8102</td><td>surgical instrument</td></tr><tr><td>Delicate straight hemostatic forceps</td><td>Roboz</td><td>RS-7114</td><td>surgical instrument</td></tr><tr><td>Delicate curved hemostatic forceps</td><td>Miltex</td><td>&amp;nbsp;7-4</td><td>surgical instrument</td></tr><tr><td>Serrefine curved, very delicate micro clamp</td><td>Roboz</td><td>RS-5471</td><td>surgical instrument</td></tr><tr><td>Micro dissecting hook</td><td>Miltex</td><td>19-220</td><td>surgical instrument</td></tr><tr><td>Angled on edge micro dissecting spring scissor</td><td>Roboz</td><td>RS-5618</td><td>surgical instrument</td></tr><tr><td>Light operating scissor</td><td>Roboz</td><td>RS-6750</td><td>surgical instrument</td></tr><tr><td>Scalpel handle and blade (#10)</td><td>Cincinnati Surgical</td><td>RS-9843</td><td>surgical instrument</td></tr><tr><td>Pack of 4-0 silk suture with curved needle</td><td>Ethicon</td><td>FS-2</td><td>surgical instrument</td></tr><tr><td>Straight micro suturing needle holder</td><td>Roboz</td><td>RS-6410</td><td>surgical instrument</td></tr><tr><td>Wound clip</td><td>Stoelting</td><td>59027</td><td>surgical instrument</td></tr><tr><td>Sterile gauze 2&quot;x2&quot;&amp;nbsp;</td><td>Dynarex</td><td>3362</td><td>consumable</td></tr><tr><td>Gauze 4&quot;x4&quot;&amp;nbsp;</td><td>Covidien</td><td>2556</td><td>consumable</td></tr><tr><td>Anesthesia vaporizer</td><td>Surgivet</td><td>V703001</td><td>equipment</td></tr><tr><td>Lucite box</td><td>Custom</td><td></td><td>equipment</td></tr><tr><td>Isoflurane</td><td>Baxter</td><td></td><td>equipment</td></tr><tr><td>Downdraft Table</td><td>Airscience</td><td></td><td>equipment</td></tr><tr><td>Table top surgical platform</td><td>Custom</td><td></td><td>equipment</td></tr><tr><td>Arterial Catheter</td><td>Custom - Scientific Commodities Inc.</td><td></td><td>consumable</td></tr><tr><td>Venous Catheter</td><td>Custom - Scientific Commodities Inc.</td><td></td><td>consumable</td></tr><tr><td>Data acquisition system</td><td>ADInstruments</td><td>Powerlab 16/30</td><td>equipment</td></tr><tr><td>Data analysis software</td><td>ADInstruments</td><td>LabChart v7.3</td><td>equipment</td></tr><tr><td>Programmable infusion pumps</td><td>Harvard Apparatus</td><td>PHD Ultra</td><td>equipment</td></tr></tbody></table>

catheterization of the carotid artery and jugular vein to perform hemodynamic measures, infusions and blood sampling in a conscious rat model

小动物模型的成功，研究重大疾病，在某种程度上，依赖模型来模拟人类状况的能力。气管内接种细菌的已知量的已被成功地用于再现肺炎，然后发育成脓毒症的发病机制。监测血液动力学参数，并提供标准的临床治疗，包括输注抗生素，​​流体和药物维持血压是至关重要的模拟在这个模型，但做常规的支持治疗有此需要动脉和静脉血管通路。该视频详细介绍了手术技术植入颈总动脉和颈总静脉导管在麻醉的老鼠。以下一个72小时的恢复期，动物将被重新麻醉，并连接到附连到所述壳体的啮齿动物相连接的植入导管的血液动力学监测系统的系绳和旋转设置。这种设置允许自由流动在研究过程中大鼠，而连续监控的压力，注入液体和药物（抗生素，升压药），并进行采血。

变化的血管压力通过流体填充导管（ 图1）传送，并转换成由血液动力波形表示的电信号（ 图2,3）。无血管通路，无法建立这些措施。波形实时流可以检测一个节拍变化击败的基础上（ 图2）和分析。压缩波形的时标允许定量发生在较长的持续时间（数分钟到数天）（ 图3，上部4的波形），它可以用在血管加压输注速率的变化相关联（ 图3，下4的波形）的变化。 <img alt="图1" src="/files/ftp_upload/51881/51881fig1.jpg" /> 图1.导管设计:在动脉和静脉导管聚氨酯，聚氨酯的部件的详细情况;。 PE，聚乙烯（段长） <a href="https://www.jove.com/files/ftp_upload/51881/51881fig1highres.jpg" target="_blank">请点击这里查看该图的放大版本。</a> <img alt="图2" src="/files/ftp_upload/51881/51881fig2.jpg" /> 图4典型的啮齿动物后7天导管在7秒2，实时地图波形。 <a href="https://www.jove.com/files/ftp_upload/51881/51881fig2highres.jpg" target="_blank">请点击此处查看该图的放大版本。</a> <img alt="图3" src="/files/ftp_upload/51881/51881fig3.jpg" /> 图3.时间压缩MAP的4个典型啮齿类（上4波形）后7天导尿波形超过1小时。当MAP被平均（下4波形），该值是用来触发ADDS高（&lt;90毫米汞柱） ，低（&lt;100毫米汞柱）或停止（&gt; 110）从输液泵的流量响应。 <a href="https://www.jove.com/files/ftp_upload/51881/51881fig3highres.jpg" target="_blank">请点击此处查看该图的放大版本。</a>

Watch this Scientific Journal Video about Catheterization of the Carotid Artery and Jugular Vein to Perform Hemodynamic Measures, Infusions and Blood Sampling in a Conscious Rat Model at JoVE.com

Catheterization of the Carotid Artery and Jugular Vein to Perform Hemodynamic Measures, Infusions and Blood Sampling in a Conscious Rat Model

Vascular accesses to measure hemodynamics, provide fluids and perform blood sampling are important to any small animal model study. We present a technique for implanting catheters into the carotid artery and the common jugular vein in an anesthetized rat for connecting to a system to perform monitoring, infusions and sampling.

颈动脉和颈静脉导管在有意识大鼠模型进行血流动力学措施，输液和血液采样

The success of a small animal model to study critical illness is, in part, dependent on the ability of the model to simulate the human condition. ...

catheterization-carotid-artery-jugular-vein-to-perform-hemodynamic

Research

JoVE Journal

Medicine

71.7K 次观看.  National Institutes of Health. Vascular accesses to measure hemodynamics, provide fluids and perform blood sampling are important to any small animal model study. We present a technique for implanting catheters into the carotid artery and the common jugular vein in an anesthetized rat for connecting to a system to perform monitoring, infusions and sampling.

视频: Catheterization of the Carotid Artery and Jugular Vein to Perform Hemodynamic Measures, Infusions and Blood Sampling in a Conscious Rat Model

Hyperinsulinemic-Euglycemic Clamp in the Conscious Rat

Type 2 diabetes (T2D) is rapidly rising in prevalence. Characterized by either inadequate insulin production or the inability to utilize insulin produced, T2D results in elevated blood glucose levels. The "gold-standard" in assessing insulin sensitivity is a hyperinsulinemic-euglycemic clamp or insulin clamp. In this procedure, insulin is infused at a constant rate resulting in a drop in blood glucose. To maintain blood glucose at a constant level, exogenous glucose (D50) is infused into the venous circulation. The amount of glucose infused to maintain homeostasis is indicative of insulin sensitivity. Here, we show the basic clamp procedure in the chronically catheterized, unrestrained, conscious rat. This model allows blood to be collected with minimal stress to the animal. Following the induction of anesthesia, a midline incision is made and the left common carotid artery and right jugular vein are catheterized. Inserted catheters are flushed with heparinized saline, then exteriorized and secured. Animals are allowed to recover for 4-5 days prior to experiments, with weight gain monitored daily. Only those animals who regain weight to pre-surgery levels are used for experiments. On the day of the experiment, rats are fasted and connected to pumps containing insulin and D50. Baseline glucose is assessed from the arterial line and used a benchmark throughout the experiment (euglycemia). Following this, insulin is infused at a constant rate into the venous circulation. To match the drop in blood glucose, D50 is infused. If the rate of D50 infusion is greater than the rate of uptake, a rise in glucose will occur. Similarly, if the rate is insufficient to match whole body glucose uptake, a drop will occur. Titration of glucose continues until stable glucose readings are achieved. Glucose levels and glucose infusion rates during this stable period are recorded and reported. Results provide an index of whole body insulin sensitivity. The technique can be refined to meet specific experimental requirements. It is further enhanced by the use of radioactive tracers that can determine tissue specific insulin-stimulated glucose uptake as well as whole body glucose turnover.

The hyperinsulinemic-euglycemic clamp is the "gold standard" for the assessment of insulin action. Insulin is infused at a constant rate stimulating glucose uptake. The amount of exogenous glucose infused to counter this drop is indicative of insulin sensitivity. Here the procedure is performed on a conscious, unrestrained rat.

在自觉大鼠高胰岛素 - 正常血糖钳

Type 2 diabetes (T2D) is rapidly rising in prevalence. Characterized by either inadequate insulin production or the inability to utilize insulin produced, ...

科研

医学

Femoral Arterial and Venous Catheterization for Blood Sampling, Drug Administration and Conscious Blood Pressure and Heart Rate Measurements

In multiple fields of study, access to the circulatory system in laboratory studies is necessary. Pharmacological studies in rats using chronically implanted catheters permit a researcher to effectively and humanely administer substances, perform repeated blood sampling and assists in conscious direct measurements of blood pressure and heart rate. Once the catheter is implanted long-term sampling is possible. Patency and catheter life depends on multiple factors including the lock solution used, flushing regimen and catheter material. This video will demonstrate the methodology of femoral artery and venous catheterization of the rat. In addition the video will demonstrate the use of the femoral venous and arterial catheters for blood sampling, drug administration and use of the arterial catheter in taking measurements of blood pressure and heart rate in a conscious freely-moving rat. A tether and harness attached to a swivel system will allow the animal to be housed and have samples taken by the researcher with minimal disruption to the animal. To maintain patency of the catheter, careful daily maintenance of the catheter is required using lock solution (100 U/ml heparinized saline), machine-ground blunt tip syringe needles and the use of syringe filters to minimize potential contamination. With careful aseptic surgical techniques, proper catheter materials and careful catheter maintenance techniques, it is possible to sustain patent catheters and healthy animals for long periods of time (several weeks).

Chronic catheterization of blood vessels in the rat is often required for administration of substances, obtain blood sample over a period of time or for direct conscious blood pressure measurements. Femoral arterial catheterization of the rat and corresponding measurements of blood pressure in the conscious animal will be demonstrated.

股动脉和静脉导管采血，药品监督管理局和意识的血压和心率测量

In multiple fields of study, access to the circulatory system in laboratory studies is necessary. Pharmacological studies in rats using chronically ...

2-Vessel Occlusion/Hypotension: A Rat Model of Global Brain Ischemia

Cardiac arrest followed by resuscitation often results in dramatic brain damage caused by ischemia and subsequent reperfusion of the brain. Global brain ischemia produces damage to specific brain regions shown to be highly sensitive to ischemia 1. Hippocampal neurons have higher sensitivity to ischemic insults compared to other cell populations, and specifically, the CA1 region of the hippocampus is particularly vulnerable to ischemia/reperfusion 2.
The design of therapeutic interventions, or study of mechanisms involved in cerebral damage, requires a model that produces damage similar to the clinical condition and in a reproducible manner. Bilateral carotid vessel occlusion with hypotension (2VOH) is a model that produces reversible forebrain ischemia, emulating the cerebral events that can occur during cardiac arrest and resuscitation. We describe a model modified from Smith et al. (1984) 2, as first presented in its current form in Sanderson, et al. (2008) 3, which produces reproducible injury to selectively vulnerable brain regions 3-6. The reliability of this model is dictated by precise control of systemic blood pressure during applied hypotension, the duration of ischemia, close temperature control, a specific anesthesia regimen, and diligent post-operative care. An 8-minute ischemic insult produces cell death of CA1 hippocampal neurons that progresses over the course of 6 to 24 hr of reperfusion, while less vulnerable brain regions are spared. This progressive cell death is easily quantified after 7-14 days of reperfusion, as a near complete loss of CA1 neurons is evident at this time.
In addition to this brain injury model, we present a method for CA1 damage quantification using a simple, yet thorough, methodology. Importantly, quantification can be accomplished using a simple camera-mounted microscope, and a free ImageJ (NIH) software plugin, obviating the need for cost-prohibitive stereology software programs and a motorized microscopic stage for damage assessment.

Bilateral carotid occlusion coupled with systemic hypotension produces global brain ischemia in the rat, resulting in damage to the hippocampus with reproducible severity. Animal subjects are impaired with predictable patterns of brain damage, they recover expediently, and mortality rates are comparatively low.

2血管闭塞/低血压：全脑缺血模型大鼠

Cardiac arrest followed by resuscitation often results in dramatic brain damage caused by ischemia and subsequent reperfusion of the brain. Global brain ...

A Model of Chronic Nutrient Infusion in the Rat

Chronic exposure to excessive levels of nutrients is postulated to affect the function of several organs and tissues and to contribute to the development of the many complications associated with obesity and the metabolic syndrome, including type 2 diabetes. To study the mechanisms by which excessive levels of glucose and fatty acids affect the pancreatic beta-cell and the secretion of insulin, we have established a chronic nutrient infusion model in the rat. The procedure consists of catheterizing the right jugular vein and left carotid artery under general anesthesia; allowing a 7-day recuperation period; connecting the catheters to the pumps using a swivel and counterweight system that enables the animal to move freely in the cage; and infusing glucose and/or Intralipid (a soybean oil emulsion which generates a mixture of approximately 80% unsaturated/20% saturated fatty acids when infused with heparin) for 72 hr. This model offers several advantages, including the possibility to finely modulate the target levels of circulating glucose and fatty acids; the option to co-infuse pharmacological compounds; and the relatively short time frame as opposed to dietary models. It can be used to examine the mechanisms of nutrient-induced dysfunction in a variety of organs and to test the effectiveness of drugs in this context.

A protocol for chronic infusions of glucose and Intralipid in rats is described. This model can be used to study the impact of nutrient excess on organ function and physiological parameters.

在大鼠模型慢性营养输液

Chronic exposure to excessive levels of nutrients is postulated to  affect the function of several organs and tissues and to contribute to the  ...

Hemodynamic Characterization of Rodent Models of Pulmonary Arterial Hypertension

Pulmonary arterial hypertension (PAH) is a rare disease of the pulmonary vasculature characterized by endothelial cell apoptosis, smooth muscle proliferation and obliteration of pulmonary arterioles. This in turn results in right ventricular (RV) failure, with significant morbidity and mortality. Rodent models of PAH, in the mouse and the rat, are important for understanding the pathophysiology underlying this rare disease. Notably, different models of PAH may be associated with different degrees of pulmonary hypertension, RV hypertrophy and RV failure. Therefore, a complete hemodynamic characterization of mice and rats with PAH is critical in determining the effects of drugs or genetic modifications on the disease. 
Here we demonstrate standard procedures for assessment of right ventricular function and hemodynamics in both rat and mouse PAH models. Echocardiography is useful in determining RV function in rats, although obtaining standard views of the right ventricle is challenging in the awake mouse. Access for right heart catheterization is obtained by the internal jugular vein in closed-chest mice and rats. Pressures can be measured using polyethylene tubing with a fluid pressure transducer or a miniature micromanometer pressure catheter. Pressure-volume loop analysis can be performed in the open chest. After obtaining hemodynamics, the rodent is euthanized. The heart can be dissected to separate the RV free wall from the left ventricle (LV) and septum, allowing an assessment of RV hypertrophy using the Fulton index (RV/(LV+S)). Then samples can be harvested from the heart, lungs and other tissues as needed.

Pulmonary arterial hypertension (PAH) is a disease of pulmonary arterioles that leads to their obliteration and the development of right ventricular failure. Rodent models of PAH are critical in understanding the pathophysiology of PAH. Here we demonstrate hemodynamic characterization, with right heart catheterization and echocardiography, in the mouse and rat.

肺动脉高压的啮齿动物模型的血流动力学特性

Pulmonary arterial hypertension (PAH) is a rare disease of the pulmonary vasculature characterized by endothelial cell apoptosis, smooth muscle ...

Simultaneous Electrocardiography Recording and Invasive Blood Pressure Measurement in Rats

For studies related to cardiovascular physiology or pathophysiology, blood pressure (BP) and electrocardiography are basic observational parameters. Research focusing on cardiovascular disease models, potential cardiovascular therapeutic targets or pharmaceutical agents requires assessment of systemic arterial pressure and heart rhythm changes. In situations where radio telemetry systems are not available or affordable, the technique of femoral artery cannulation is an alternative way to obtain intra-arterial pressure waveform recordings and systemic BP measurements. This technique is economical and can be performed with standard equipment in animal facilities. However, invasive arterial pressure recording requires cannulation of small arteries, which can be a challenging surgical skill. Here, we present step-by-step protocols for femoral artery cannulation procedures. Key procedures include the calibration of the data acquisition system, tissue dissection and femoral artery cannulation, and setup of the arterial cannulation system for pressure recording. Surface electrocardiography recording procedures are also included. We also present examples of BP recordings from normotensive and hypertensive rats. This protocol allows reliable direct recordings of systemic BP with simultaneous electrocardiography.

Here, we describe a setup for simultaneous recording of electrocardiography and intra-arterial blood pressure (BP) in experimental rats, which can be done with standard equipment in animal facilities and can be applied to physiological or pharmacological studies to investigate pathogenic or therapeutic mechanisms in cardiovascular medicine.

大鼠同时心电图记录及侵入性血压测量

For studies related to cardiovascular physiology or pathophysiology, blood pressure (BP) and electrocardiography are basic observational parameters. ...

Continuous Blood Sampling in Small Animal Positron Emission Tomography/Computed Tomography Enables the Measurement of the Arterial Input Function

For quantitative analysis and bio-kinetic modeling of positron emission tomography/computed tomography (PET/CT) data, the determination of the temporal blood time-activity concentration also known as arterial input function (AIF) is a key point, especially for the characterization of animal disease models and the introduction of newly developed radiotracers. The knowledge of radiotracer availability in the blood helps to interpret PET/CT-derived data of tissue activity. For this purpose, online blood sampling during the PET/CT imaging is advisable to measure the AIF. In contrast to manual blood sampling and image-derived approaches, continuous online blood sampling has several advantages. Besides the minimized blood loss, there is an improved resolution and a superior accuracy for the blood activity measurement. However, the major drawback of online blood sampling is the costly and time-consuming preparation to catheterize the femoral vessels of the animal. Here, we describe an easy and complete workflow for catheterization and continuous blood sampling during small animal PET/CT imaging and compared it to manual blood sampling and an image-derived approach. Using this highly-standardized workflow, the determination of the fluorodeoxyglucose ([18F]FDG) AIF is demonstrated. Further, this procedure can be applied to any radiotracer in combination with different animal models to create fundamental knowledge of tracer kinetic and model characteristics. This allows a more precise evaluation of the behavior of pharmaceuticals, both for diagnostic and therapeutic approaches in the preclinical research of oncological, neurodegenerative and myocardial diseases.

Here a protocol for continuous blood sampling during PET/CT imaging of rats to measure the arterial input function (AIF) is described. The catheterization, the calibration and setup of the system and the data analysis of the blood radioactivity are demonstrated. The generated data provide input parameters for subsequent bio-kinetic modeling.

小型动物正电子发射断层扫描/计算机断层扫描中的连续血液采样支持动脉输入函数的测量

For quantitative analysis and bio-kinetic modeling of positron emission tomography/computed tomography (PET/CT) data, the determination of the temporal ...

Establishment of Deep Hypothermic Circulatory Arrest in Rats

Deep hypothermic circulatory arrest (DHCA) is routinely applied during surgeries for complex congenital heart disease and aortic arch disease. The present study aims to provide a method for establishing DHCA in rats. To evaluate the impact of the DHCA process on vital signs, a normal temperature cardiopulmonary bypass (CPB) rat model without circulatory arrest was used as a control. As expected, DHCA led to a significant decrease in body temperature and mean arterial blood pressure. The blood gas analysis indicated that DHCA increased lactic acid levels but did not influence the blood pH and the concentrations of hemoglobin, hematocrit, Na+, Cl&#8722;, K+, and glucose. Furthermore, compared with the normal temperature CPB rats, the results of the transmission electron microscopy showed a mild increase in hippocampal autophagosomes in the DHCA rats.

This protocol presents the establishment of deep hypothermic circulatory arrest in rats, which can be applied to investigate systemic inflammatory response syndrome, ischemia/reperfusion injury, oxidative stress, neuroinflammation, etc.

在大鼠中建立深低温循环停滞

Deep hypothermic circulatory arrest (DHCA) is routinely applied during surgeries for complex congenital heart disease and aortic arch disease. The present ...

免疫与感染

Microsurgical Skills of Establishing Permanent Jugular Vein Cannulation in Rats for Serial Blood Sampling of Orally Administered Drug

Blood sampling in small laboratory animals is necessary for pharmaceutical lead optimization but can cause great harm and stress to experimental animals, which could potentially affect results. The jugular vein cannulation (JVC) in rats is a widely used model for repeated blood collection but requires adequate training of surgery skills and animal care. This article details the microsurgical procedures for establishing and maintaining a permanent JVC rat model with specific focus on the placement and sealing of the jugular cannula. The importance of monitoring physiological (e.g., body weight, food, and water intake) and hematological parameters, was highlighted with results presented for 6 days post-surgery during the rat&#39;s recovery. The drug-plasma concentration-time profile of orally administered natural phenol ellagic acid was determined in the JVC rat model.

Detailed microsurgical techniques are demonstrated to establish a longer-term jugular vein cannulation rat model for sequential blood collection in the same animal. Physiological and hematological parameters have been monitored during the rat's recovery phase. This model has been applied to study pharmacokinetics of orally administered polyphenol without inducing animal stress.

建立大鼠永久性颈静脉插管的显微外科技能，用于口服药物的连续血液采样

Blood sampling in small laboratory animals is necessary for pharmaceutical lead optimization but can cause great harm and stress to experimental animals, ...

锁骨下静脉穿刺法对清醒大鼠重复采血

Subclavian Vein Blood Sampling in Conscious Rats

There are several established methods for obtaining repeated blood samples from rats, with the most commonly employed methods being lateral tail vein sampling without anesthesia and jugular vein sampling with anesthesia. However, most of these methods require assistance and anesthetic equipment and sometimes pose difficulties in terms of blood collection or the poor quality of blood samples. In addition, these methods of blood collection consume significant time and human resources when repeated blood sampling is required for a large number of rats. This study presents a technique for repetitive blood sampling in non-anesthetized rats by a single proficient individual. Highly satisfactory blood samples can be obtained by puncturing the subclavian vein. The method demonstrated an impressive overall success rate of 95%, with a median time of merely 2 min from rat restraint to the completion of blood collection. Furthermore, performing consecutive blood collections within the designated range does not inflict any harm on the rats. This method is worth promoting for blood collection, especially in large-scale pharmacokinetic studies.

作者聚焦:在非麻醉大鼠中开发一种高效的血液采集方法 

Here we present a combination of effective rat restriction and subclavian vein puncture methods that enable rapid, safe, and repeated blood collection in rats without anesthesia.

有意识大鼠的锁骨下静脉血液采样

There are several established methods for obtaining repeated blood samples from rats, with the most commonly employed methods being lateral tail vein ...