This work was supported by funding from the March of Dimes FY12-133 (S.K.E.) and the National Institutes of Health R01HD037831 (S.K.E.). We would like to thank Dr. Deborah Frank for critical review of the manuscript.

注:符合动物提出了美国国立卫生研究院的维护和使用指南所有动物的程序。所有协议批准了动物研究委员会在圣路易斯华盛顿大学。 注:特殊材料和设备来获得本协议所列材料和设备表。 雌性大鼠1.定时育种<ol><li>品种两岁六个月的成年女性与男性有2小时的时间窗口。特定育种小时是不需要的，只要它们保持在实验的全部相一致。 </li><li>通过交配插头的存在证实怀孕。指定的日期和人力资源上的插头第一看作是一天后性交0（dpc0）。启动压力录制时，记得具体的时间。 </li><li>在dpc8-13进行手术。确切的DPC将取决于小鼠品系。 </li><li>如果要判断一只老鼠怀孕手术前，PE与连接到成像系统管理麻醉前一个40 MHz的线阵探头腹部rform超声成像。这将允许妊娠囊的可视化。如果一个超声系统不可用，使用鼠标的重量。 注:dpc0和dpc8和腹胀之间至少有2克体重增加是怀孕的良好指标。 </li></ol> 2.遥测手术准备<ol><li>使用高压锅术前24小时内消毒器械，纱布，棉签，手术单，和枪头。 </li><li>清洁手术表面用70％的乙醇。 </li><li>打开加热垫（专门为啮齿动物）和珠灭菌。 </li></ol> 3.遥测手术<ol><li>麻醉，用4-5％异氟烷以1-2升/分钟的氧流量。 </li><li>等到鼠标显示麻醉的迹象，然后注入皮下丁丙诺啡（镇痛:每20克0.1毫升）。 </li><li>剃鼠标从肋的底部的腹部向下至膀胱。 </li><li>把鼠标上的预温加热垫的手术台，并覆盖鼠标的眼睛眼药膏，以防止干燥。 </li><li>开始无菌技术。戴上无菌手套，并保持包装的无菌表面上要放置碘和酒精擦洗。 </li><li>从包中取出或盐水的遥测并用无菌镊子将其放置在无菌纱布。 <ol><li>使用注射器（由遥测供应商提供的）来填充导管尖端与凝胶。轻轻挤压导管创造足够的负压吸引凝胶进入导管。 </li></ol></li><li>广场上的吸管无菌尖端。吸管2微升的手术胶水，并设置吸管上的无菌表面。 </li><li>清洁外科手术部位，用碘，然后用70％乙醇擦拭的面积。 </li><li>切手术悬垂足够大的一个洞对于所需的切口和地点在鼠标区域。 </li><li>执行脚捏（踏板响应），以确保该鼠标正在一个切口之前完全麻醉。 </li><li>做一个小的垂直正中切口。接下来，通过底层体壁的肌肉类似的切口。 </li><li>轻轻一拉出来子宫体腔，找到包含最幼崽是可行子宫角。 <ol><li>可选 - 对于另外的miRNA，插入遥测前注入病毒载体进入子宫肌层。 </li></ol></li><li>做一个小切口，在子宫角尖为3 mm的尖端弹簧剪刀。 </li><li>螺纹导管，而不挤压，子宫壁和胎儿囊之间。插入导管过去几囊，以减少其意外脱落。确保胎囊不会被插入的干扰。 </li><li>吸取所述手术胶水以遥测插入部位附着的导管与uterine喇叭和防止导管滑出子宫角。等待几秒钟，胶水变得僵硬，然后小心地将子宫后面的体腔内。 </li><li>放置在体腔内的遥测在导管插入部位的相反侧。 </li><li>缝合体腔与​​6.0可吸收缝合线封闭。首先，做一个荷包缝合，然后三到四个单独的针。 </li><li>缝合皮肤通过使用相同的技术，但是用6-0非可吸收的缝线。 </li><li>棉签丁卡因（外用止痛药）软膏在切口部位。 </li><li>注射0.3-0.4毫升无菌生理盐水皮下，以帮助补充水分的动物。 </li><li>监测手术后的动物，直到它完全清醒。不要将鼠标背部在笼子里，直到它能够自由地保持其头部上面的表格为某些类型的床上用品可以窒息重度麻醉鼠标。对于鼠标已经麻醉30-45分钟，不要指望日Ë鼠标是由1小时和15分钟完全清醒。 </li><li>如果进行额外的手术，请用70％的乙醇和珠再次使用前消毒器械。 </li></ol> 4，手术后恢复<ol><li>监控日志的日常手术后的进展。目录体重增加和损失，并注意有无嗜睡，出血，或手术切口闭合不全。 <ol><li>喂老鼠软化小鼠食物（〜苹果酱的一致性）或专门回收的减肥食品。 </li><li>检查缝线，并确保他们不拉出，但尽量不要处理鼠标过分。 </li></ol></li></ol> 5.遥测记录<ol><li>感兴趣的日子（两到三天后，至少在鼠标已经从手术恢复），放置一个磁铁靠近鼠标打开遥测。把鼠标笼上的接收器。接收器上的光应打开，如果遥测被激活。 </li><li>校准全新teleme<html> TER值与制造商特定的校准提供。重复使用遥测校准规范将被保存在软件。 </li><li>在软件，开始记录右击"动物"，然后点击"开始采样，持续，"这使得采样在高达500 Hz的速率。确保软件设置保存和跟踪。 注:当采样鼠标图标变成绿色，压力被记录。在分娩过程中干扰鼠标可以中断或改变劳动，所以观察谨慎。 </li><li>经过利益输送或时间段，停止录制右击"动物"，然后选择"停止录音，所有"。用磁铁关闭遥测，以节省电池寿命，因为遥测电池通常只持续1.5个月的连续记录。 </li><li>安乐死鼠标的CO 2过量和恢复的遥测。幼崽被斩首处死。 </li></ol>TLE"&gt; 6。遥测灭菌<ol><li>将遥测中的1％的酶洗涤剂在去离子水中过夜，在室温下的溶液中。 </li><li>用去离子水冲洗的遥测，并删除所有组织和胶水，小心不要挤导管。 </li><li>孵育在2％戊二醛的遥测为24小时，在室温下进行消毒。 </li><li>冲洗用无菌生理盐水的遥测，并存储在无菌盐水中，直到下一次注入或将被发送回给制造商进行翻新。如果组织位于导管内灭菌后遥测应该翻新。通常情况下遥测可以无需翻新之前使用4-5次。 </li></ol>

<ol>
	<li>Slattery, M. M., Morrison, J. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Preterm+delivery.">Preterm delivery.</a> Lancet. 360, 1489-1497 (2002).</li><li>Goldenberg, R. L., Culhane, J. F., Iams, J. D., Romero, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Epidemiology+and+causes+of+preterm+birth.">Epidemiology and causes of preterm birth.</a> Lancet. 371, 75-84 (2008).</li><li>Keelan, J. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pharmacological+inhibition+of+inflammatory+pathways+for+the+prevention+of+preterm+birth.">Pharmacological inhibition of inflammatory pathways for the prevention of preterm birth.</a> Journal Of Reproductive Immunology. 88, 176-184 (2011).</li><li>Voltolini, C., 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=Understanding+spontaneous+preterm+birth:+from+underlying+mechanisms+to+predictive+and+preventive+interventions.">Understanding spontaneous preterm birth: from underlying mechanisms to predictive and preventive interventions.</a> Reproductive Sciences. 20, 1274-1292 (2013).</li><li>Ratajczak, C. K., Muglia, L. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Insights+into+parturition+biology+from+genetically+altered+mice.">Insights into parturition biology from genetically altered mice.</a> Pediatric research. 64, 581-589 (2008).</li><li>Rajakumar, A., 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=Placental+HIF-1+alpha,+HIF-2+alpha,+membrane+and+soluble+VEGF+receptor-1+proteins+are+not+increased+in+normotensive+pregnancies+complicated+by+late-onset+intrauterine+growth+restriction.">Placental HIF-1 alpha, HIF-2 alpha, membrane and soluble VEGF receptor-1 proteins are not increased in normotensive pregnancies complicated by late-onset intrauterine growth restriction.</a> American Journal Of Physiology. Regulatory, Integrative And Comparative Physiology. 293, (2007).</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.+American+journal+of+physiology.">Comparison of simultaneous measurement of mouse systolic arterial blood pressure by radiotelemetry and tail-cuff methods. American journal of physiology.</a> Heart And Circulatory Physiology. 286, H2408-H2415 (2004).</li><li>Mackay, L. B., Shi, L. B., Maul, H. .., Maner, W. L., Garfield, R. 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+effect+of+bilateral+pelvic+neurectomy+on+cervical+ripening+in+pregnant+rats.">The effect of bilateral pelvic neurectomy on cervical ripening in pregnant rats.</a> Journal Of Perinatal Medicine. 37, 263-269 (2009).</li><li>Pierce, S. L., Kutschke, W., Cabeza, R., England, S. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=In+vivo+measurement+of+intrauterine+pressure+by+telemetry:+a+new+approach+for+studying+parturition+in+mouse+models.">In vivo measurement of intrauterine pressure by telemetry: a new approach for studying parturition in mouse models.</a> Physiol Genomics. 42, 310-316 (2010).</li><li>Imamura, T., Luedke, C. E., Vogt, S. K., Muglia, L. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Oxytocin+modulates+the+onset+of+murine+parturition+by+competing+ovarian+and+uterine+effects.">Oxytocin modulates the onset of murine parturition by competing ovarian and uterine effects.</a> American Journal Of Physiology. Regulatory, Integrative And Comparative Physiology. 279, R1061-R1067 (2000).</li><li>McCloskey, C., 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=The+inwardly+rectifying+K++channel+KIR7.1+controls+uterine+excitability+throughout+pregnancy.">The inwardly rectifying K+ channel KIR7.1 controls uterine excitability throughout pregnancy.</a> EMBO Mol Med. 6 (9), 1161-1174 (2014).</li></ol>

The authors have nothing to disclose.

子宫肌层收缩的研究一直依靠肌肉紧张体外测量。这种方法可用于新开发的子宫收缩药物不能被施用给活的动物的早期检测是有用的。然而， 在体内的方法是必要的，以纵向理解劳动的进 ​​展。 体内测量一举数得。首先，它们使过怀孕期间变化子宫收缩图案的完整画面的捕获。其次，他们消除体外录音要求子宫组织的偏压被垂直或水平地安装，限制了我们的子宫的?...

早产是围产儿发病率和死亡率在发达国家的主要原因;它负责50％围产期发病率及围产儿死亡率1,2 75％。早产是多方面的，可能是特发性。虽然许多研究已经出现在分子和电通路将来自一个静止组织肌层成收缩1，早产的确切病理生理学仍然难以捉摸。内分泌，炎性和基因调控都被链接到早产3,4。然而，伦理问题限制于早产在人体中的机制进行研究的能力。 鉴于这些限制，许多研究人员已经转向小鼠作为模型系统来研究分娩的生理学。小鼠具有短怀孕持续约3个星期，并且可以很容易地遗传操作。此外，多个基因小鼠模型被开发，以确定信号通路，这对于劳动5是必不可少的。尽管小鼠和人类分娩之间的关键差异，小鼠和人类有许多是必不可少的劳动相同的机制，包括宫内炎症和感染6。因此，老鼠作为一个宝贵的工具用于检查子宫活动。迄今为止，金标准测定小鼠子宫收缩已经离体等长张力的录音;然而，这被限制在每个实验1妊娠时间点，并且需要切除子宫从它的生理环境。另一个重要因素是需要大量的对这些研究的动物。最后，这种方法不允许进行纵向研究审查引产和早产病理生理学。 在用于研究动脉压辐射遥测设备取得的最新进展在老鼠和INT改变7大鼠8 rauterine压力乞求问题相同的技术是否可以用于怀孕期间，研究改变小鼠子宫内的压力。最初的故障排除后，已成功开发出一种方法来衡量引产和进展的鼠标。这种体内方法可以测量从静态子宫指示有力宫缩的高压状态的低压力状态的过渡。这种方法也已经能够检测到与受损分娩和野生型小鼠的转基因小鼠之间显著压力差，表明基因表达的9的生理影响。这个协议提供了一个实时的， 在体内方法来研究怀孕期间小鼠子宫的压力。

<table><tbody><tr><td>Surgical Glue</td><td>3M Vetbond</td><td>1469SB</td><td>Tissue Adhesive--stored in Drierite</td></tr><tr><td>Absorbable 6-0 Sutures</td><td>Ethicon</td><td>J212H</td><td>Vicryl</td></tr><tr><td>Non-absorbable 6-0 Sutures</td><td>Ethicon</td><td>8706H</td><td>Prolene</td></tr><tr><td>Water Jacket Blanket + Heating Pad</td><td>Gaymer</td><td>T/Pump PN 11184-000 Blanket-66N111CC</td><td>Specialized for rodents</td></tr><tr><td>Bead sterilizer</td><td>Keller</td><td>Z378577</td><td>Steri 250 Sterilizer</td></tr><tr><td>Disecting Microscope</td><td>Nikon</td><td>SMZ754</td><td>Fibre optic gooseneck external light source</td></tr><tr><td>Sterile Surgical Gloves--Latex&amp;nbsp;</td><td>Cardinal Health Triflex</td><td>2D7253</td><td></td></tr><tr><td>PhysioTel PA-C10 Pressure Transmitter</td><td>&amp;nbsp;Data Sciences International</td><td>270-0135-001</td><td>TA11PA-C10</td></tr><tr><td>Telemeter Reciever</td><td>&amp;nbsp;Data Sciences International</td><td>272-6001-001</td><td>RPC-1</td></tr><tr><td>Dataquest ART 4.3.2 Analysis Platinum</td><td>&amp;nbsp;Data Sciences International</td><td>271-0147-141</td><td>Analysis software</td></tr><tr><td>Diet Recovery Gel</td><td>Clear H2O</td><td>72-01-5022</td><td>Purified Soft Diet for Rodents</td></tr><tr><td>Tergazyme</td><td>Sigma-Aldrich</td><td>Z273287</td><td>Enzyme detergent</td></tr></tbody></table>

intrauterine telemetry to measure mouse contractile pressure in vivo

分子和电信号的复杂的集成是需要变换静止子宫成收缩的器官在怀孕的末端。尽管子宫收缩的关键调节剂的发现，这个过程还没有完全理解。转基因小鼠提供在学习分娩的理想模型。以前，以研究子宫收缩在小鼠的唯一方法是离体等长张力的录音，这是不理想的有几个原因。子宫必须从它的生理环境被除去，调查在有限的时间当然是可能的，并且小鼠必须牺牲。辐射遥测的最新发展已经允许纵向，实时体内小鼠宫腔压力的测量。在这里，宫内遥测植入到分娩的描述来测量从妊娠中期的小鼠子宫压力变化。在记者的差异比较野生型和转基因小鼠之间URES，感兴趣的基因的生理影响可以阐明。该技术应加快用于妊娠期间治疗子宫肌层病症的治疗剂，包括早产的发展。

实时体内宫内压力可通过使用遥测采集系统进行记录。搭配鼠标同步录像压取样来捕捉每个小狗的传递的准确时间和相关交货时间，母亲的子宫内的压力。所有的数据点就可以在整个记录作图以产生压力-时间曲线图（ 图1A）。在妊娠期间这个图显示的各点时，宫内压力变化以及暗光周期之间的压力差。此外，宫内压力的移动平均可以绘制（ 图1B）。这（分娩过程?...

Watch this Scientific Journal Video about Intrauterine Telemetry to Measure Mouse Contractile Pressure In Vivo at JoVE.com

Intrauterine Telemetry to Measure Mouse Contractile Pressure In Vivo

This manuscript provides a protocol for implanting telemeters in the mouse for the purpose of measuring intrauterine pressures during pregnancy.

宫内遥测来衡量鼠标收缩压力<em&gt;在体内</em

A complex integration of molecular and electrical signals is needed to transform a quiescent uterus into a contractile organ at the end of pregnancy. ...

intrauterine-telemetry-to-measure-mouse-contractile-pressure-in-vivo

Research

JoVE Journal

Medicine

Intrauterine Telemetry to Measure Mouse Contractile Pressure In Vivo

9.6K 次观看.  Washington University in St. Louis. This manuscript provides a protocol for implanting telemeters in the mouse for the purpose of measuring intrauterine pressures during pregnancy. 

视频: Intrauterine Telemetry to Measure Mouse Contractile Pressure In Vivo

Measuring Left Ventricular Pressure in Late Embryonic and Neonatal Mice

Blood pressure increases significantly during embryonic and postnatal development in vertebrate animals. In the mouse, blood flow is first detectable around embryonic day (E) 8.51. Systolic left ventricular (LV) pressure is 2 mmHg at E9.5 and 11 mmHg at E14.52. At these mid-embryonic stages, the LV is clearly visible through the chest wall for invasive pressure measurements because the ribs and skin are not fully developed. Between E14.5 and birth (approximately E21) imaging methods must be used to view the LV. After birth, mean arterial pressure increases from 30 - 70 mmHg from postnatal day (P) 2 - 353. Beyond P20, arterial pressure can be measured with solid-state catheters (i.e. Millar or Scisense). Before P20, these catheters are too big for developing mouse arteries and arterial pressure must be measured with custom pulled plastic catheters attached to fluid-filled pressure transducers3 or glass micropipettes attached to servo null pressure transducers4.
Our recent work has shown that the greatest increase in blood pressure occurs during the late embryonic to early postnatal period in mice5-7. This large increase in blood pressure may influence smooth muscle cell (SMC) phenotype in developing arteries and trigger important mechanotransduction events. In human disease, where the mechanical properties of developing arteries are compromised by defects in extracellular matrix proteins (i.e. Marfan's Syndrome8 and Supravalvular Aortic Stenosis9) the rapid changes in blood pressure during this period may contribute to disease phenotype and severity through alterations in mechanotransduction signals. Therefore, it is important to be able to measure blood pressure changes during late embryonic and neonatal periods in mouse models of human disease.
We describe a method for measuring LV pressure in late embryonic (E18) and early postnatal (P1 - 20) mice. A needle attached to a fluid-filled pressure transducer is inserted into the LV under ultrasound guidance. Care is taken to maintain normal cardiac function during the experimental protocol, especially for the embryonic mice. Representative data are presented and limitations of the protocol are discussed.

Measuring left ventricular pressure (LV) in embryonic and neonatal mice is described. Pressure is measured by inserting a needle connected to a fluid-filled transducer into the LV under ultrasound guidance. Care must be taken to maintain normal cardiac function during the experimental protocol.

测量中晚期胚胎和新生小鼠左心室压力

Blood pressure increases significantly during embryonic and postnatal development in vertebrate animals. In the mouse, blood flow is first detectable ...

科研

生物工程

Implantation of Radiotelemetry Transmitters Yielding Data on ECG, Heart Rate, Core Body Temperature and Activity in Free-moving Laboratory Mice

The laboratory mouse is the animal species of choice for most biomedical research, in both the academic sphere and the pharmaceutical industry. Mice are a manageable size and relatively easy to house. These factors, together with the availability of a wealth of spontaneous and experimentally induced mutants, make laboratory mice ideally suited to a wide variety of research areas.
In cardiovascular, pharmacological and toxicological research, accurate measurement of parameters relating to the circulatory system of laboratory animals is often required. Determination of heart rate, heart rate variability, and duration of PQ and QT intervals are based on electrocardiogram (ECG) recordings. However, obtaining reliable ECG curves as well as physiological data such as core body temperature in mice can be difficult using conventional measurement techniques, which require connecting sensors and lead wires to a restrained, tethered, or even anaesthetized animal. Data obtained in this fashion must be interpreted with caution, as it is well known that restraining and anesthesia can have a major artifactual influence on physiological parameters1, 2.
Radiotelemetry enables data to be collected from conscious and untethered animals. Measurements can be conducted even in freely moving animals, and without requiring the investigator to be in the proximity of the animal. Thus, known sources of artifacts are avoided, and accurate and reliable measurements are assured. This methodology also reduces interanimal variability, thus reducing the number of animals used, rendering this technology the most humane method of monitoring physiological parameters in laboratory animals3, 4. Constant advancements in data acquisition technology and implant miniaturization mean that it is now possible to record physiological parameters and locomotor activity continuously and in realtime over longer periods such as hours, days or even weeks3, 5.
Here, we describe a surgical technique for implantation of a commercially available telemetry transmitter used for continuous measurements of core body temperature, locomotor activity and biopotential (i.e. onelead ECG), from which heart rate, heart rate variability, and PQ and QT intervals can be established in freeroaming, untethered mice. We also present pre-operative procedures and protocols for post-operative intensive care and pain treatment that improve recovery, well-being and survival rates in implanted mice5, 6.

A surgical technique for implantation of commercially available telemetry transmitters used for continuous measurement of biopotential (one-lead ECG), heart rate, core body temperature and locomotor activity in freely moving mice is shown. Recommendations and protocols for post-operative care and pain relief, improving recovery, well being and survival rate are also presented.

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

The laboratory mouse is the animal species of choice for most biomedical research, in both the academic sphere and the pharmaceutical industry. Mice are a ...

医学

In utero Measurement of Heart Rate in Mouse by Noninvasive M-mode Echocardiography

Congenital heart disease (CHD) is the most frequent noninfectious cause of death at birth. The incidence of CHD ranges from 4 to 50/1,000 births (Disease and injury regional estimates, World Health Organization, 2004). Surgeries that often compromise the quality of life are required to correct heart defects, reminding us of the importance of finding the causes of CHD. Mutant mouse models and live imaging technology have become essential tools to study the etiology of this disease. Although advanced methods allow live imaging of abnormal hearts in embryos, the physiological and hemodynamic states of the latter are often compromised due to surgical and/or lengthy procedures. Noninvasive ultrasound imaging, however, can be used without surgically exposing the embryos, thereby maintaining their physiology. Herein, we use simple M-mode ultrasound to assess heart rates of embryos at E18.5 in utero. The detection of abnormal heart rates is indeed a good indicator of dysfunction of the heart and thus constitutes a first step in the identification of developmental defects that may lead to heart failure.

In utero Measurement of Heart Rate in Mouse by Noninvasive M-mode Echocardiography

Ultra-high frequency ultrasound is a powerful live imaging tool to examine cardiac abnormalities in small animals. Its noninvasiveness allows maintaining the physiologic state of embryos. Herein, we show the use of M-mode ultrasound to measure the heart rates of embryos at E18.5 in utero.

心率在小鼠中通过无创M型超声心动图测量子宫内

Congenital heart disease (CHD) is the most frequent noninfectious cause of death at birth. The incidence of CHD ranges from 4 to 50/1,000 births (Disease ...

Mouse Maternal Autonomic Changes from Pregnancy to Pup-Rearing

Analysis of Electrocardiograms and Behavior in Mice from Pregnancy to Lactation Period

Changes in the mother-offspring relationship are presumably accompanied by dynamic changes in the autonomic nervous system. Although temporal measurements of autonomic activity have been performed in human mothers and infants, the analysis of long-term changes remains unexplored. Mouse mothers can form social bonds with their pups and have a short period of pregnancy and lactation, which makes them useful for the examination of physiological changes from pregnancy to pup-rearing. Therefore, a telemetry system was used for several weeks to measure the changes in the autonomic nervous system and the behavior of mouse mothers. The current results showed that an electrocardiogram (ECG) could be stably recorded regardless of the movements of mothers and parturition. ECG analysis showed that the heart rate gradually decreased from pregnancy to lactation, and sympathetic activity sharply increased as the pups developed. Furthermore, the simultaneous recording of behavior and ECG in the home cage enabled us to understand the behavior-dependent influences on the ECG, thereby revealing the characteristics of autonomic nervous activity during each behavior. Thus, the present experimental method helps to understand how the physiological characteristics of mothers change from pregnancy through pup rearing, supporting the healthy development of pups.

A simultaneous recording of autonomic activity and detailed maternal behavior of mother mice from pregnancy to lactation was achieved using a telemetry system. This method helps to understand the dynamics of the physiological and behavioral characteristics in mothers from pregnancy to weaning.

Changes in the mother-offspring relationship are presumably accompanied by dynamic changes in the autonomic nervous system. Although temporal measurements ...

行为学

Long-term Blood Pressure Measurement in Freely Moving Mice Using Telemetry

During the development of new vasoactive agents, arterial blood pressure monitoring is crucial for evaluating the efficacy of the new proposed drugs. Indeed, research focusing on the discovery of new potential therapeutic targets using genetically altered mice requires a reliable, long-term assessment of the systemic arterial pressure variation. Currently, the gold standard for obtaining long-term measurements of blood pressure in ambulatory mice uses implantable radio-transmitters, which require&#160;artery cannulation. This technique eliminates the need for tethering, restraining, or anesthetizing the animals which introduce stress and artifacts during data sampling. However, arterial blood pressure monitoring in mice via catheterization can be rather challenging due to the small size of the arteries. Here we present a step-by-step guide to illustrate the crucial key passages for a successful subcutaneous implantation of radio-transmitters and carotid artery cannulation in mice. We also include examples of long-term blood pressure activity taken from freely moving mice after a period of post-surgery recovery. Following this procedure will allow reliable direct blood pressure recordings from multiple animals simultaneously.

The goal of this protocol is to assess systemic blood pressure in conscious freely moving mice using implantable radio-telemetry devices.

自由活动小鼠使用遥测长期血压测量

During the development of new vasoactive agents, arterial blood pressure monitoring is crucial for evaluating the efficacy of the new proposed drugs. ...

Application of Consistent Massage-Like Perturbations on Mouse Calves and Monitoring the Resulting Intramuscular Pressure Changes

Massage is generally recognized to be beneficial for relieving pain and inflammation. Although previous studies have reported anti-inflammatory effects of massage on skeletal muscles, the molecular mechanisms behind are poorly understood. We have recently developed a simple device to apply local cyclical compression (LCC), which can generate intramuscular pressure waves with varying amplitudes. Using this device, we have demonstrated that LCC modulates inflammatory responses of macrophages in situ and alleviates immobilization-induced muscle atrophy. Here, we describe protocols for the optimization and application of LCC as a massage-like intervention against immobilization-induced inflammation and atrophy of skeletal muscles of mouse hindlimbs. The protocol that we have developed can be useful for investigating the mechanism underlying beneficial effects of physical exercise and massage. Our experimental system provides a prototype of the analytical approach to elucidate the mechanical regulation of muscle homeostasis, although further development needs to be made for more comprehensive studies.

Here we describe the protocols for applying defined mechanical loads to mouse calves and for monitoring the concomitant intramuscular pressure changes. The experimental systems that we have developed can be useful for investigating the mechanism behind the beneficial effects of physical exercise and massage.

在小鼠小腿上应用一致的按摩样扰动，并监测由此产生的肌肉压力变化

Massage is generally recognized to be beneficial for relieving pain and inflammation. Although previous studies have reported anti-inflammatory effects of ...

免疫与感染

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. ...

Real-Time Electrocardiogram Monitoring During Treadmill Training in Mice

Regular physical exercise is a major contributor to cardiovascular health, influencing various metabolic as well as electrophysiological processes. However, in certain cardiac diseases such as inherited arrhythmia syndromes, e.g., arrhythmogenic cardiomyopathy (ACM) or myocarditis, physical exercise may have negative effects on the heart leading to a proarrhythmogenic substrate production. Currently, the underlying molecular mechanisms of exercise-related proarrhythmogenic remodeling are largely unknown, thus it remains unclear which frequency, duration, and intensity of exercise can be considered safe in the context of disease(s).
The proposed method allows to study proarrhythmic/antiarrhythmic effects of physical exercise by combining treadmill training with real-time monitoring of the ECG. Implantable telemetry devices are used to continuously record the ECG of freely moving mice over a period of up to 3 months both at rest and during treadmill training. Data acquisition software with its analysis modules is used to analyze basic ECG parameters such as heart rate, P wave duration, PR interval, QRS interval, or QT duration at rest, during and after training. Furthermore, heart rate variability (HRV) parameters and occurrence of arrhythmias are evaluated. In brief, this manuscript describes a step-by-step approach to experimentally explore exercise induced effects on cardiac electrophysiology, including potential proarrhythmogenic remodeling in mouse models.

Electrocardiogram (ECG) is the key variable to understanding cardiac electrophysiology. Physical exercise has beneficial effects but may also be harmful in the context of cardiovascular diseases. This manuscript provides a method of recording real-time ECG during exercise, which can serve to investigate its effects on cardiac electrophysiology in mice.

小鼠跑步机训练期间的实时心电图监测

Regular physical exercise is a major contributor to cardiovascular health, influencing various metabolic as well as electrophysiological processes. ...

Ex Vivo Method for Assessing the Mouse Reproductive Tract Spontaneous Motility and a MATLAB-based Uterus Motion Tracking Algorithm for Data Analysis

Dysmenorrhea, or painful cramping, is the most common symptom associated with menses in females and its severity can hinder women&#39;s everyday lives. Here, we present an easy and inexpensive method that would be instrumental for testing new drugs decreasing uterine contractility. This method utilizes the unique ability of the entire mouse reproductive tract to exhibit spontaneous motility when maintained ex vivo in a Petri dish containing oxygenated Krebs buffer. This spontaneous motility resembles the wave-like myometrial activity of the human uterus, referred to as endometrial waves. To demonstrate the effectiveness of the method, we employed a well-known uterine relaxant drug, epinephrine. We demonstrate that the spontaneous motility of the entire mouse reproductive tract can be quickly and reversibly inhibited by 1 &#181;M epinephrine in this Petri dish model. Documenting the changes of uterine motility can be easily done using an ordinary smart phone or a sophisticated digital camera. We developed a MATLAB-based algorithm allowing motion tracking to quantify spontaneous uterine motility changes by measuring the rate of uterine horn movements. A major advantage of this ex vivo approach is that the reproductive tract remains intact throughout the entire experiment, preserving all intrinsic intrauterine cellular interactions. The major limitation of this approach is that up to 10-20% of uteri may exhibit no spontaneous motility. Thus far, this is the first quantitative ex vivo method for assessing spontaneous uterine motility in a Petri dish model.

Uterine contractions are important for the well-being of females. However, pathologically increased contractility may result in dysmenorrhea, especially in younger females. Here, we describe a simple ex vivo preparation allowing quick assessment of the efficacy of smooth muscle relaxants that may be used for treating dysmenorrhea.

用于评估小鼠生殖道自发性的方法和用于数据分析的基于 MATLAB 的 Uterus 运动跟踪算法

Dysmenorrhea, or painful cramping, is the most common symptom associated with menses in females and its severity can hinder women&#39;s everyday lives. ...

生物学

Continuous Telemetric In Utero Tracheal Pressure Measurements in Fetal Lambs

Normal in utero lung development and growth rely upon the expansion of airspaces and the controlled efflux of lung liquid into the amniotic space. Infants with congenital diaphragmatic hernia (CDH) also have lung hypoplasia due to occupation of the chest cavity by the stomach and bowel and, in the most severe cases, the liver. Balloon tracheal occlusion reduces the severity of lung hypoplasia in fetuses with CDH but increases the risk of premature birth. Understanding the optimal occlusion pressure and duration required to improve lung hypoplasia with tracheal occlusion is essential to improving in utero corrective treatments for CDH. The study reports a new method for continuous measurement of the intratracheal and amniotic pressures in an unoccluded and occluded fetal lamb surgical model of CDH. Time-pregnant Merino ewes underwent two recovery hysterotomies: the first at ~80 days of gestation to create the CDH, and the second at ~101 days of gestation to occlude the fetal trachea and implant an intratracheal and amniotic pressure measurement device. Lambs were delivered at ~142 days, and the pressure measurement device was removed and cleaned. The data were downloaded and filtered using a 6 h window. Transrespiratory pressure was calculated.

<meta charset="utf8"></head><body>胎羔羊子宫内气管压力连续遥测测量

Optimization of occlusion pressure and duration required to improve lung hypoplasia with tracheal occlusion is essential to improve in utero corrective treatments for congenital diaphragmatic hernia (CDH). This study reports a new method for continuous measurement of the tracheal pressure in an occluded and unoccluded fetal lamb surgical model of CDH.

胎羔羊子宫内气管压力连续遥测测量

Normal in utero lung development and growth rely upon the expansion of airspaces and the controlled efflux of lung liquid into the amniotic space. Infants ...

宫内遥测来衡量鼠标收缩压力

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