植入的无线电遥测发射器产生心电图,心率,身体核心温度和自由移动的实验室小鼠的活动上的数据

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Summary

显示一个市售遥测发射机,用于连续测量生物电位植入手术技术(一导联心电图),心率,体温和自由活动的小鼠自发活动。建议和手术后的保健和缓解疼痛的协议,改善恢复,福祉和成活率也。

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Cesarovic, N., Jirkof, P., Rettich, A., Arras, M. Implantation of Radiotelemetry Transmitters Yielding Data on ECG, Heart Rate, Core Body Temperature and Activity in Free-moving Laboratory Mice. J. Vis. Exp. (57), e3260, doi:10.3791/3260 (2011).

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Abstract

实验小鼠的选择对于大多数生物医学研究的动物物种在学术领域和制药业,。小鼠是一个可管理的大小和相对容易的房子。这些因素,加上丰富的自发性和实验诱导的突变体的可用性,非常适合于各种各样的研究领域的实验室小鼠。

在心血管,药理学和毒理学研究,实验动物的循环系统有关参数的精确测量往往是必需的。心率,心率变异性,PQ和QT间隔时间的确定是根据心电图(ECG)录音。然而,获得可靠的心电图曲线,以及如在小鼠的核心体温等生理数据,可以使用传统的测量技术,这就要求传感器和导线连接内敛,系绳,甚至一个难以aesthetized动物。以这种方式获得的数据必须谨慎解释,因为它是众所周知的,限制和麻醉可以有一个重要生理参数1,2 artifactual影响。

无线电遥测使意识和不受限制的动物从收集到的数据。在自由移动的动物,甚至可以进行测量,并没有要求研究者在邻近的动物。因此,避免了文物的来源,并保证准确和可靠的测量。这种方法还降低了interanimal变异,从而减少使用动物的数量,使这一技术的生理参数监测实验动物3,4,最人道的方法。在数据采集技术和植入微型化常数进步意味着它现在可以连续和实时记录不再p生理参数和自主活动eriods如小时,天或什至3,5。

在这里,我们描述了一个市售的核心体温,自主活动和生物电位(即onelead心电图),心率,心率变异性,和PQ和QT间期可以连续测量使用的遥测发射器的植入手术技术成立于freeroaming,不受限制的小鼠。我们也出席了会议前手术和手术后的重症监护和疼痛治疗,改善恢复协议,福祉和在植入小鼠的5,6存活率。

Protocol

动物实验已批准由州兽医办公室(瑞士苏黎世)。房屋和实验过程与瑞士动物保护法的规定,并符合欧洲指令保护动物用于科学目的(欧洲议会和理事会2010年9月22日的指令2010/63/EU)。

1。手术前的考虑

1.1小鼠:住房需求,一般情况和健康监测

这是建议,从供应商提供,或从外部灭鼠殖民地转移的小鼠在手术前至少两周抵达住房设施。这期间应该允许的动物,以适应新的环境和设施的具体的住房条件。小鼠,作为社会生活的动物,应该被安置在兼容的群体,在这个适应期。用于监控一个人的水平食物和水的消耗,每个鼠标采用术前3天的单发射器植入手术后,直到10天。建立遥测发射机荷瘤小鼠的时间线如图1所示。这是非常重要的动物,在良好的健康和手术条件。因此,在手术前,动物应进行监测,每天一次为2-3天,一般情况(外观,姿态,自发的行为),以及体重,食物和水的消耗。这些数据记录在病历(一般状况和健康监测数据表,表1)建立个别的基准水平的一般状况和整体健康和福祉。从实验中,应排除任何动物疾病或手术前的一般状况受损的症状。

1.2在手术前一天的头发剪裁

植入前一天,以剃除ANIMALS手术,小鼠麻醉简要地在一个小的(8x8x8cm)有机玻璃室使用纯氧(600毫升/分钟)“七氟醚(8%)或异氟醚(5%)。翻正反射的损失后,鼠标室,前颈部和腹毛动物趴在背靠着裁剪;麻醉维持与鼻面罩与七氟醚3-4%或约5分钟异氟醚1.5-3%纯氧在600毫升/分钟的流速。修剪头发后,动物允许唤醒,然后带回自己的家笼。

2。植入

2.1工作环境,遥测发射机的准备

植入当天,所有关于发射准备和手术的程序进行工作台上配备手术显微镜层流罩。无菌条件下保证蒸压仪器和设备的使用经济需求和消毒的材料,并进行消毒工作台上7。植入前,遥测发射机(ETA - 10,数据科学国际,圣保罗,明尼苏达,美国)先准备。从他们的无菌包装中取出后,发射器的引线缩短到适当的长度被植入老鼠的大小。在多数成人远交或近交系小鼠中,红色的电极可缩短到约42毫米,长度约55毫米的白色/无色电极。绝缘管从远端(感觉)部分的信息删除:约20毫米的管道是从红色的电极删除,约10毫米的管道是从白色/无色电极。每个电极的远端部分(这是现在没有管),形成一个循环固定绢缝合(PERMA Handseide,爱惜康,日本东京,德国6-0,)结束。准备电极后,发射器是放置在西澳RM无菌生理盐水准备时被植入动物麻醉和手术的准备。

2.2麻醉

在5-10分钟吸入麻醉诱导前,咪达唑仑(4毫克/千克)和芬太尼(0.04毫克/千克)的混合物皮下注射作为术前用药,从而提供了镇静和先发制人的镇痛。一般吸入麻醉诱导放置在感应室内的动物和引进的挥发性麻醉剂(七氟醚8%纯氧600毫升/分钟的异氟醚5%)。当动物显示它转移到工作台层流罩下,并在背靠着放置在一个特别设计的金属用鼻子面具和油管从麻醉仪器安装板的翻正反射的损失。麻醉维持自主呼吸(七氟醚在600毫升/分钟的流量的3-4%或异氟醚1.5-3%纯氧)。在麻醉过程中,动物的眼睛S是保护软膏(维生素A,Baush博士伦,Steinhausen,瑞士)。虽然躺在金属板动物是由水浴加热表面温热(39℃+ / -1)的工作台上。

2.3手术

用70%乙醇消毒皮肤前颈部和腹部地区。一个在皮肤1 - 1.5厘米长的切口是沿腹部中线较低的胸部。负(白色/无色)导致皮下隧道,从胸部到颈部,一个小切口(≤0.5厘米),是在纵向方向。皮肤和相关组织准备,以使固定电极丝循环的空间。位于气管右侧的肌肉之间的电线回路是固定的,使用两个绢缝合(PERMA Handseide,6-0,爱惜康,日本东京,德国)。伤口在脖子上,然后关闭与可吸收缝合线(6-0薇乔,爱惜康,日本东京,德国)层。然后打开腹壁白线和身体遥测发射机放置到鼠标的腹腔。缝合剑突之间的肝脏和膈肌左上腹地区(图2)在于它用这样的方式丝绸缝合线的正极(红色)电极循环。然后,腹部肌肉层封闭,可吸收缝合线(6-0薇乔,爱惜康,日本东京,德国)。最后才关闭腹壁,Sulfadoxin和甲氧苄啶的混合物[(30毫克/千克和6毫克/公斤,分别溶于1毫升的生理盐水(0.9%)和体温约(38-39 ° C)]最后,注入腹腔抗感染的预防的目的,并支持流体的动态平衡。腹部皮肤恢复主食(精确,3米保健,圣保罗,明尼苏达,美国)。

3。手术后的护理

手术和麻醉完成后,0.1毫克/公斤丁丙诺啡(Temgesic,埃塞克斯化学公司,瑞士卢塞恩)和5毫克/公斤美洛昔康(Metacam,勃林格殷格翰,巴塞尔,瑞士)是用于治疗疼痛的管理皮下,和动物留下的温水(39℃+ / -1)工作台上收回约2H表面。连同缓解疼痛(每天两次:丁丙诺啡,0.1毫克/千克,美洛昔康5毫克/公斤),300μL葡萄糖(5%)和300μL生理盐水(0.9%),加热到体温的支持疗法组成,适用于皮下注射两次4天。进一步复苏的支持,这是值得的动物提供了一个额外的饮水瓶含15%葡萄糖溶液。在4-10天的恢复期,这是建议,动物保持温暖。因此,在我们的例子中,小鼠被安置在变暖柜(30℃+ / - 1)。监测的一般状况和体重,以及食物和水的消耗,每天进行一次,根据一般条件和手术后10天的健康监测数据表(表1)。人性化的端点,即一种动物的牺牲,以避免不必要的痛苦和疼痛,如果恢复进展情况并不理想,在下列情况下实现:

  1. 如果全身情况较差,即动物大幅萎靡(运动后没有被感动/推),其体表发冷,尽管气候变暖,动物应立即euthanatized。
  2. 如果发射器植入后第4天,动物表演的冷漠的明显迹象,是非常积极的,或不显示任何食物的摄入量,应该立即euthanatized。
  3. 发射器植入后第8天,动物比较前手术后的日子,以显示在体重明显增加此外,它消耗在l东手术前每日食物摄入量的80%。如果不符合这些条件之一是,动物应立即euthanatized。

在植入后10天,动物调回标准的住房条件下的动物房。老鼠应该被安置在兼容的群体,让社会的互动,以避免长期的个人住房,它可以有重大影响,对后续实验8,9的读出的不利影响。小鼠发射器植入后的第一个实验是进行数据采集开始前至少4个星期的疗养期间。

4。数据采集

数据收集是通过触摸动物用磁铁发起,届时发送器。 Dataquest的最先进的软件(国际数据科学,圣保罗,明尼苏达,美国)坐标检测,收集,分析和GRaphical演示(以波的形式的形式),从一个或更多的动物的信号。收购方案收集数据信号发送到电脑,通过数据交换矩阵(国际数据科学)的转换器和接收器。这个程序可以连续收集特定长度的时间定期或样品的数据,并保存在计算机的硬盘驱动器的数据。由于发射信号的范围和质量在很大程度上取决于笼的材料组成及周边设备(如金属和塑料),建议接收板置于尽可能接近动物,如根据动物的笼或以上的实验区,如实验台或跑步机上。这是建议的记录和数据传输系统的正确配置一个在连续采样模式下的实时测量的短检查检查。已收集和存储数据后,他们可以积特德,上市,并分析了各种不同的参数,使用分析程序。录音系统(例如定义取样手法),和分析软件(如心脏心率变异性参数,PQ间期和QT间建立生物电势/心电图曲线)的配置的详细信息,可以找到制造商的手册。生物识别的规划和统计方法的遥测数据采集和解释有用的有价值的提示是在别处发表了3。

5。代表性的成果:

一个整体计划所描述的过程如图1所示。植入的发射器的位置,包括电极的位置获得的心脏(一导联心电图)biopotentials,如图2所示。原始数据的例子,从短期的生物电势曲线(单导联心电图),和长期心率,体温和个人自主活动的录音小鼠,分别在图3和图4。图5给出了从长期测量后,实验组小鼠的出版,的数据的一个例子。从biopotentials曲线可以建立其他几个参数。介绍心率变异性参数5,QT间期和PQ 间隔 10 的例子 ,11别处发表。

表1。一般情况和健康监测数据表 ,点击这里下载此模板有利于个别鼠标的一般状况和健康监测工作表。。植入手术前必须建立基线检查动物的外表,姿势,和自发的行为,以及体重的决心,食物和水的消耗,每天一次,连续3天。获得的基线测定比较每天手术后10天的服务,以评估手术后恢复的进展。此外,手术后的护理和治疗疼痛的医疗记录的形式是有据可查的。人性化的端点上的说明,以便决定鼠标是否应该牺牲,以防止不必要的痛苦和折磨,如果动物不符合的标准,植入后的快速恢复。

图1
图1。建立遥测发射机荷瘤小鼠的附表。按时间顺序显示鼠标可以用于实验和数据采集的时间点的发射器植入有关的程序。

图2
图2。X光片/草图显示的位置植入telemetRY发射机。变送器的主体定位在腹腔。积极的主导作用,形成成丝环和固定缝合剑突过程。负极从颈部胸部皮下隧道和固定线回路之间的直接气管旁边的肌肉。 X光片是取自作者在以前出版实验动物9。

图3
图3生物电势曲线。一导联心电图曲线从有意识的鼠标和七氟醚吸入麻醉下同一动物的原始打印输出。心率遥测系统自动计算。在麻醉状态下记录3秒的序列表示的心率为440 BPM。在有意识的鼠标记录的曲线显示了心率为660 BPM,属于心脏率在预期范围内,在模式率疏导或饮食等体育活动。从生物电势/一导联心电图曲线,心率变异性参数,interbeat间隔,和PQ和QT间可以建立与制造商的软件使用。

图4
图4:在健康和患病小鼠的长期测量的原始数据。心率(BPM),核心体温(℃)和自发活动(计数)测量小鼠单独住在他们家笼没有任何人或实验程序的干扰。心率记录为30秒,每5分钟(采样频率1000赫兹)。核心体温为10秒,每5分钟采样。自主活动的连续记录和储存在5分钟的时间间隔。五分钟的数据点跟踪为6.5天。从三只老鼠具有不同的遥测测量记录身体状况。健康的老鼠显示了明显的昼夜节律与正常生理值和自发活动的行为,在黑暗(夜)相增加。相比之下,大手术后,心率增加,特别是在日光下阶段,自主活动是郁闷。第三个鼠标患有肿瘤的慢性疾病,其心率和体温的昼夜节律出现夷为平地,自主活动减少。取自作者Altex 12以前出版的心率测量有代表性的数据(正常价值和大手术后)。

图5
图5。长期遥测测量结果后,实验演示示例图是取自作者在以前出版实验动物1。作为示范实验,50分钟isoflurane或七氟醚麻醉。麻醉药长期心率,体温和后动物自发活动的影响比较清醒。使用16个发射器植入小鼠,遥测数据在8%的麻醉小鼠记录,而动物是单设,并允许自由地徜徉在他们的家笼。对于长期postanesthetic影响的分析,我们考虑到了,价值观以来主要在夜间活动的老鼠在一个24小时的周期大大不同。因此,每个动物的遥测值的方法分别计算夜间(12小时黑暗)和天(12 h光照)相。设立了一个单独的正常价值计算从3天前麻醉手段。麻醉后的每一天,意味着黑暗与光明的阶段相比,个人的正常价值,导致增量值。因此,增量值代表正常数值的偏差(前成立麻醉)在相应的12小时昼夜时间。列代表平均8只;横道标准偏差。星号表示的意义在P≤0.05(单向方差分析比较组是指在正常价值的麻醉后每4天)。

Discussion

无线电遥测是一个功能强大的替代传统的方法在生物医学研究中的生理参数的测量。植入式发射器,接收器和数据采集和分析的硬件和软件组成的高品质的遥测系统,现在市面上,即使是像老鼠的小动物。遥测是唯一的技术,目前的数据收集从奔放,自由活动的小鼠。通过使用这种方法,它现在可以不断地和/或更长的时间,从动物居住在自己熟悉的环境,从而最大限度地减少应激对动物和随之而来的实验文物收集数据。形式和位置的信息进行了优化,以获得信号即使在快速运动(如挣扎,跑步,战斗),或在一个直立的姿势 9 。因此,精确的测量,可以在实验中获得,如在麻醉过程中,强调在duction,而在跑步机上运行,​​在感染实验中,和许多其他的实验情况,在行为实验。

然而,为了获得可靠,重复性好,无瑕疵的数据,它是至关重要的,以排除环境影响,和我们特别提请注意标准条件的重要性。这是建议,是从电子和噪音,包括超声波的声音,特别敏感,小鼠隔离室。此外,没有任何干扰,如游客或无关的实验程序,应允许时进行测量。为了避免干扰的影响(特别是在家庭笼测量的情况下),一切必要的饲养程序,应在房间开始每次测量之前完成。此外,小鼠的住房,尤其是当男性团体或个别可以对测量的影响,必须加以考虑,当解放军nning实验9。此外,老鼠必须健康,无鼠病原体,因为潜在的或明显的感染,以及疾病或任何其他健康损害的,可以有相当大的影响生理参数和活动行为。因此,小鼠完全康复后植入,并给予足够的时间来适应轴承开始之前的任何实验发射机。

小鼠无线电遥测数据收集需要的遥测发射机的初步手术植入。这应该是只能由训练有素的人员与手术技巧,以尽量减少组织损伤和随后的疼痛和痛苦。实验者的最基本甚至最先进的(微)手术技巧,建议在新鲜尸体鼠标使用培训种植体(即,假人,由制造商提供)建立的程序和熟悉与细节执行的第一次试验这一类手术。经过这样的训练,大多数实验者将能够植入这种成功的发射器的类型和几个植入后会达到一个有用的能力。

在手术过程中应保持无菌条件下,保持低感染的微生物的负担和风险。然而,不能提供完整的无菌因为一些具体的,不育冲突的条件,在小鼠(例如,广泛的发型剪裁和消毒的冷却效果,不切实际的绷带保护伤口)。因此,抗感染的预防是在植入管理。适合的镇痛治疗和明确界定的监测计划以及足够的术后护理,发挥至关重要的作用在实验中取得令人满意的结果。

总体而言,在小鼠的遥测发射机手术植入动物的压力。特别是,如果在规范中的基因改造IFIC鼠标线影响的表型和损害动物的身体状况,在围手术期的时限并发症和死亡率增加,植入后可能的风险。为了避免不必要的痛苦,个人表现不理想恢复或长期疗养应该被释放从实验和牺牲才到达一个垂死的阶段。为此,一个数据表(表1:一般条件和健康监测数据表)促进症状的关键系统的监测和人文端点提供意见已经确立。因此,恢复被记录在病历或检验杂志,这使得这种方法(即植入手术和手术后的恢复)有关当局和动物福利机构,负责动物实验(如透明导电风格, IACUC)。

Disclosures

没有利益冲突的声明。

Acknowledgements

作者想感谢德国查尔斯河提供CD - 1小鼠。我们也感谢罗宾施耐德和中央支持住房小鼠的生物实验室的工作人员。我们诚挚感谢优秀的技术援助和教授库尔特Burki动植物李启慷慨提供研究设施和资源。

References

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  3. Kramer, K., Kinter, L. B. Evaluation and applications of radiotelemetry in small laboratory animals. Physiol. Genomics. 13, 197-205 (2003).
  4. Kramer, K. Use of telemetry to record electrocardiogram and heart rate in freely moving mice. J. Pharmacol. Toxicol. Methods. 30, 209-215 (1993).
  5. Arras, M., Rettich, A., Cinelli, P., Kasermann, H. P., Burki, K. Assessment of post-laparotomy pain in laboratory mice by telemetric recording of heart rate and heart rate variability. BMC. Vet. Res. 3, 16-16 (2007).
  6. Schuler, B., Rettich, A., Vogel, J., Gassmann,, Arras, M. Optimized surgical techniques and postoperative care improve survival rates and permit accurate telemetric recording in exercising mice. BMC. Vet. Res. 5, 28-28 (2009).
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  8. Rettich, A., Kasermann, H. P., Pelczar, P., Burki, K., Arras, M. The physiological and behavioral impact of sensory contact among unfamiliar adult mice in the laboratory. J. Appl. Anim. Welf. Sci. 9, 277-288 (2006).
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Erratum

Formal Correction: Erratum: Implantation of Radiotelemetry Transmitters Yielding Data on ECG, Heart Rate, Core Body Temperature and Activity in Free-moving Laboratory Mice
Posted by JoVE Editors on 10/09/2016. Citeable Link.

Corrections in the Protocol and Discussion sections have been made to: Implantation of Radiotelemetry Transmitters Yielding Data on ECG, Heart Rate, Core Body Temperature and Activity in Free-moving Laboratory Mice

Step 1.2 in the Protocol has been updated from:

1.2 Hair clipping at one day prior to surgery

The day prior to implantation, in order to shave the animals for surgery, mice are anesthetized briefly in a small (8x8x8cm) Perspex chamber using sevoflurane (8%) or isoflurane (5%) in pure oxygen (600 mL/min). After loss of the righting reflex, the mouse is taken out of the chamber and the anterior neck and abdominal hair is clipped with the animal lying in dorsal recumbence; anesthesia is maintained for approximately 5 minutes with a nose mask with sevoflurane 3-4% or isoflurane 1.5-3% in pure oxygen at a flow rate of 600 mL/min. After clipping the hair, the animals are allowed to awaken and are then brought back to their home cage.

to:

After the last health check or directly prior surgery, in order to shave the animals for surgery, mice are anesthetized briefly in a small (8x8x8cm) Perspex chamber using sevoflurane (8%) or isoflurane (5%) in pure oxygen (600 mL/min). Shaving the animals one day before surgery prevents hair stubbles in the operating field. After loss of the righting reflex, the mouse is taken out of the chamber and the anterior neck and abdominal hair is clipped with the animal lying in dorsal recumbence; anesthesia is maintained for approximately 5 minutes with a nose mask with sevoflurane 3-4% or isoflurane 1.5-3% in pure oxygen at a flow rate of 600 mL/min. After clipping the hair, the animals are allowed to awaken and are then brought back to their home cage.

Step 2.3 in the Protocol has been updated from:

2.3 Surgery

The skin of the anterior neck and abdominal region is disinfected with 70% ethanol. A 1- to 1.5-cm-long incision in the skin is made from the lower thorax along the midline to the abdomen. The negative (white/colourless) lead is tunnelled subcutaneously from the thorax to the neck, where a small incision (≤0.5 cm) is made in the longitudinal direction. The skin and underlying tissues are prepared to make space for the fixation of the wire loop of the electrode. The wire loop is fixed between the muscles located to the right of the trachea, using two thin silk sutures (PERMA-Handseide, 6-0, Ethicon, Norderstedt, Germany). The wound in the neck is then closed with absorbable sutures (VICRYL 6-0, Ethicon, Norderstedt, Germany) in layers. The abdominal wall is then opened at the linea alba and the body of the telemetric transmitter is placed into the abdominal cavity of the mouse. The wire loop of the positive (red) electrode is sutured to the xiphoid process with silk sutures in such a way that it lies between the liver and the diaphragm in the left upper abdominal region (Figure 2). Then, the muscle layers of the abdominal region are closed with absorbable sutures (VICRYL 6-0, Ethicon, Norderstedt, Germany). Before finally closing the abdominal wall, a mixture of Sulfadoxin and Trimethoprim [(30 mg/kg and 6 mg/kg, respectively; dissolved in 1 mL of saline (0.9%) and at approximately body temperature (38-39°C)] is injected into the abdominal cavity for the purposes of anti-infective prophylaxis and to support fluid homeostasis. Finally, the skin of the abdominal region is restored with staples (Precise, 3 M Health Care, St. Paul, MN, USA).

to:

2.3 Surgery

The skin of the anterior neck and abdominal region is disinfected for 5 minutes with 70% ethanol, chlorhexidine or iodine using a soaked cotton swap. A 1- to 1.5-cm-long incision in the skin is made from the lower thorax along the midline to the abdomen. The negative (white/colourless) lead is tunnelled subcutaneously from the thorax to the neck, where a small incision (≤0.5 cm) is made in the longitudinal direction. The skin and underlying tissues are prepared to make space for the fixation of the wire loop of the electrode. The wire loop is fixed between the muscles located to the right of the trachea, using two thin silk sutures (PERMA-Handseide, 6-0, Ethicon, Norderstedt, Germany). The wound in the neck is then closed with absorbable sutures (VICRYL 6-0, Ethicon, Norderstedt, Germany) in layers. The abdominal wall is then opened at the linea alba and the body of the telemetric transmitter is placed into the abdominal cavity of the mouse. The wire loop of the positive (red) electrode is sutured to the xiphoid process with silk sutures in such a way that it lies between the liver and the diaphragm in the left upper abdominal region (Figure 2). Then, the muscle layers of the abdominal
region are closed with absorbable sutures (VICRYL 6-0, Ethicon, Norderstedt, Germany). Before finally closing the abdominal wall, a mixture of Sulfadoxin and Trimethoprim [(30 mg/kg and 6 mg/kg, respectively; dissolved in 1 mL of saline (0.9%) and at approximately body temperature (38-39°C)] is injected into the abdominal cavity for the purposes of anti-infective prophylaxis and to support fluid homeostasis. Finally, the skin of the abdominal region is restored with staples (Precise, 3 M Health Care, St. Paul, MN, USA) or intracutaneous, running, absorbable sutures (VICRYL 6-0, Ethicon, Norderstedt, Germany).

Step 3 in the Protocol has been updated from:

3. Post-operative care

After completion of surgery and anesthesia, 0.1 mg/kg of buprenorphine (Temgesic, Essex Chemie AG, Lucerne, Switzerland) and 5 mg/kg of meloxicam (Metacam, Boehringer Ingelheim, Basel, Switzerland) is administered subcutaneously for pain treatment, and the animals are left on the warm (39°C +/-1) surface of the work bench to recover for approximately 2h. Together with pain relief (twice daily: buprenorphine, 0.1 mg/kg and meloxicam 5 mg/kg), supportive therapy consisting of 300 μL glucose (5%) and 300 μL saline (0.9%) warmed to body temperature, is applied subcutaneously twice daily for 4 days. For further recovery support, it is worthwhile providing the animals with an additional drinking bottle containing 15% glucose solution During the recovery period of 4-10 days, it is recommended that the animals are kept warm. Therefore, in our case, the mice are housed in a warming cabinet (30°C +/- 1). Monitoring of general condition and body weight, as well as food and water consumption, is performed once daily according to the general condition and health monitoring data sheet (Table 1) for 10 days post-operatively. Humane endpoints, i.e. the sacrifice of an animal to avoid unnecessary suffering and pain if progression of recovery is unsatisfactory, are realised under the following conditions:

i. If in poor general condition, i.e. the animal is substantially apathetic (no movement after being touched/pushed) and its body surface feels cold despite warming, the animal should be euthanatized immediately

ii. If, on day 4 after transmitter implantation, the animal shows clear signs of apathy, is extremely aggressive or does not show any food intake, it should be euthanatized immediately.

iii. On day 8 after transmitter implantation, the animal has to display a clear increase in body weight in comparison to the preceding post-operative days. Moreover, it has to consume at least 80% of the pre-operative daily food intake. If one of these conditions is not met, the animal should be euthanatized immediately.

At 10 days after implantation, the animal is transferred back to the animal room under standard housing conditions. Mice should be housed in compatible groups to allow social interaction and to prevent the adverse effects of long-term individual housing, which can have substantial impacts on the read-out of subsequent experiments8, 9. Mice should have a period of at least 4 weeks convalescence after transmitter implantation before the first experiment is conducted and data acquisition begins.

to:

After completion of surgery and anesthesia, 0.1 mg/kg of buprenorphine (Temgesic, Essex Chemie AG, Lucerne, Switzerland) and 5 mg/kg of meloxicam (Metacam, Boehringer Ingelheim, Basel, Switzerland) is administered subcutaneously for pain treatment, and the animals are left on the warm (39°C +/-1) surface of the work bench to recover for approximately 2h. Together with pain relief (twice daily: buprenorphine, 0.1 mg/kg and meloxicam 5 mg/kg), supportive therapy consisting of 300 μL glucose (5%) and 300 μL saline (0.9%) warmed to body temperature, is injected subcutaneously twice daily for 4 days. For further recovery support, it is worthwhile providing the animals with an additional drinking bottle containing 15% glucose solution During the recovery period of 4-10 days, it is recommended that the animals are kept warm. Therefore, in our case, the mice are housed in a warming cabinet (30°C +/- 1). Monitoring of general condition and body weight, as well as food and water consumption, is performed once daily according to the general condition and health monitoring data sheet (Table 1) for 10 days post-operatively. Humane endpoints, i.e. the sacrifice of an animal to avoid unnecessary suffering and pain if progression of recovery is unsatisfactory, are realised under the following conditions:

i. If in poor general condition, i.e. the animal is substantially apathetic (no movement after being touched/pushed) and its body surface feels cold despite warming, the animal should be euthanatized immediately

ii. If, on day 4 after transmitter implantation, the animal shows clear signs of apathy, is extremely aggressive or does not show any food intake, it should be euthanatized immediately.

iii. On day 8 after transmitter implantation, the animal has to display a clear increase in body weight in comparison to the preceding post-operative days. Moreover, it has to consume at least 80% of the pre-operative daily food intake. If one of these conditions is not met, the animal should be euthanatized immediately.

At 10 days after implantation, the animal is transferred back to the animal room under standard housing conditions. In case staples have been used, these should be removed 7-10 days after surgery; absorbable sutures have not to be removed. Mice should be housed in compatible groups to allow social interaction and to prevent the adverse effects of long-term individual housing, which can have substantial impacts on the read-out of subsequent experiments8, 9. Mice should have a period of at least 4 weeks convalescence after transmitter implantation before the first experiment is conducted and data acquisition begins.

The 4th paragraph in the Discussion has been updated from:

Aseptic conditions should be maintained during surgery to keep the microbiological burden and the risk of infections low. However, complete sterility cannot be provided because of some specific, sterility conflicting conditions in mice (e.g., cooling effect of extensive hair clipping and disinfection, impracticality of bandages to protect the wounds). Thus, anti-infective prophylaxis is administered during the implantation. Well tailored analgesic treatment and a clearly defined monitoring plan as well as adequate post-operative care play a crucial role in the satisfactory outcome of the experiment.

to:

Aseptic conditions should be maintained during surgery to keep the microbiological burden and the risk of infections low. However, if there are doubts that asepsis was breached because of some specific, sterility conflicting conditions in mice (e.g., cooling effect of extensive hair clipping and disinfection, impracticality of bandages to protect the wounds). Anti-infective prophylaxis should be administered during the implantation. Well-tailored analgesic treatment and a clearly defined monitoring plan as well as adequate post-operative care play a crucial role in the satisfactory outcome of the experiment.

Comments

1 Comment

  1. With the help of radio telemetry systems we are in the state of gathering the data about various animals. By the application of radio telemetry we can measure ECG, heart rate etc.

    http://www.stiengineering.com.au/industries/industrial-automation-and-scada-systems.aspx

    Reply
    Posted by: Alann P.
    August 5, 2013 - 2:57 AM

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