Name: optimization of transesophageal atrial pacing to assess atrial fibrillation susceptibility in mice
Uploaded: 2022-06-29T15:00:00
Description: Watch this Scientific Journal Video about Optimization of Transesophageal Atrial Pacing to Assess Atrial Fibrillation Susceptibility in Mice at JoVE.com

图 2 是使用 BioRender.com 创建的。这项工作得到了美国国立卫生研究院国家心肺血液研究所（HL096844和HL133127）的资助;美国心脏协会（2160035，18SFRN34230125和903918[MBM]）;以及美国国立卫生研究院国家转化科学促进中心（UL1 TR000445）。

该程序已获得范德比尔特机构动物护理和使用委员会的批准，并且与实验动物护理和使用指南一致。该方案是使用遗传9 和获得的10 （例如高血压）小鼠AF易感性模型开发的。操作员对所研究小鼠的表型视而不见。
1. 动物选择
<ol><li>对于遗传模型，如下所述每两周（即每隔一周）对小鼠进行心房起搏（参见步骤6），以确定AF易...

<ol>
	<li>Sumitomo, 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=Association+of+atrial+arrhythmia+and+sinus+node+dysfunction+in+patients+with+catecholaminergic+polymorphic+ventricular+tachycardia.">Association of atrial arrhythmia and sinus node dysfunction in patients with catecholaminergic polymorphic ventricular tachycardia.</a> Circulation Journal. 71 (10), 1606-1609 (2007).</li><li>Fukui, 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=Role+of+leptin+signaling+in+the+pathogenesis+of+angiotensin+II-mediated+atrial+fibrosis+and+fibrillation.">Role of leptin signaling in the pathogenesis of angiotensin II-mediated atrial fibrosis and fibrillation.</a> Circulation: Arrhythmia and Electrophysiology. 6 (2), 402-409 (2013).</li><li>Schutter, D., 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=Animal+models+of+atrial+fibrillation.">Animal models of atrial fibrillation.</a> Circulation Research. 127 (1), 91-110 (2020).</li><li>Li, 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=Ryanodine+receptor-mediated+calcium+leak+drives+progressive+development+of+an+atrial+fibrillation+substrate+in+a+transgenic+mouse+model.">Ryanodine receptor-mediated calcium leak drives progressive development of an atrial fibrillation substrate in a transgenic mouse model.</a> Circulation. 129 (12), 1276-1285 (2014).</li><li>Wakimoto, H., 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=Induction+of+atrial+tachycardia+and+fibrillation+in+the+mouse+heart.">Induction of atrial tachycardia and fibrillation in the mouse heart.</a> Cardiovascular Research. 50 (3), 463-473 (2001).</li><li>Schrickel, J. W., 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=Induction+of+atrial+fibrillation+in+mice+by+rapid+transesophageal+atrial+pacing.">Induction of atrial fibrillation in mice by rapid transesophageal atrial pacing.</a> Basic Research in Cardiology. 97 (6), 452-460 (2002).</li><li>Verheule, S., 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=Increased+vulnerability+to+atrial+fibrillation+in+transgenic+mice+with+selective+atrial+fibrosis+caused+by+overexpression+of+TGF-beta1.">Increased vulnerability to atrial fibrillation in transgenic mice with selective atrial fibrosis caused by overexpression of TGF-beta1.</a> Circulation Research. 94 (11), 1458-1465 (2004).</li><li>Faggioni, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Suppression+of+spontaneous+ca+elevations+prevents+atrial+fibrillation+in+calsequestrin+2-null+hearts.">Suppression of spontaneous ca elevations prevents atrial fibrillation in calsequestrin 2-null hearts.</a> Circulation: Arrhythmia and Electrophysiology. 7 (2), 313-320 (2014).</li><li>Murphy, M. 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=Optimizing+transesophageal+atrial+pacing+in+mice+to+detect+atrial+fibrillation.">Optimizing transesophageal atrial pacing in mice to detect atrial fibrillation.</a> American Journal of Physiology - Heart and Circulatory Physiology. 332 (1), 36-43 (2022).</li><li>Prinsen, J. K., 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=Highly+reactive+isolevuglandins+promote+atrial+fibrillation+caused+by+hypertension.">Highly reactive isolevuglandins promote atrial fibrillation caused by hypertension.</a> JACC: Basic to Translational Science. 5 (6), 602-615 (2020).</li><li>Aschar-Sobbi, R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Increased+atrial+arrhythmia+susceptibility+induced+by+intense+endurance+exercise+in+mice+requires+TNF&#945;.">Increased atrial arrhythmia susceptibility induced by intense endurance exercise in mice requires TNF&#945;.</a> Nature Communications. 6, 6018 (2015).</li><li>Bruegmann, T., 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=Optogenetic+termination+of+atrial+fibrillation+in+mice.">Optogenetic termination of atrial fibrillation in mice.</a> Cardiovascular Research. 114 (5), 713-723 (2017).</li><li>Matsushita, 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=IL-1&#946;+plays+an+important+role+in+pressure+overload-induced+atrial+fibrillation+in+mice.">IL-1&#946; plays an important role in pressure overload-induced atrial fibrillation in mice.</a> Biological and Pharmaceutical Bulletin. 42 (4), 543-546 (2019).</li><li>Sato, S., 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=Cardiac+overexpression+of+perilipin+2+induces+atrial+steatosis,+connexin+43+remodeling,+and+atrial+fibrillation+in+aged+mice.">Cardiac overexpression of perilipin 2 induces atrial steatosis, connexin 43 remodeling, and atrial fibrillation in aged mice.</a> American Journal of Physiology - Endocrinology and Metabolism. 317 (6), 1193-1204 (2019).</li><li>Li, N., Wehrens, X. H. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Programmed+electrical+stimulation+in+mice.">Programmed electrical stimulation in mice.</a> Journal of Visualized Experiments. (39), e1730 (2010).</li><li>Yao, 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=Enhanced+cardiomyocyte+NLRP3+inflammasome+signaling+promotes+atrial+fibrillation.">Enhanced cardiomyocyte NLRP3 inflammasome signaling promotes atrial fibrillation.</a> Circulation. 138 (20), 2227-2242 (2018).</li><li>Purohit, 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=Oxidized+Ca2+/calmodulin-dependent+protein+kinase+II+triggers+atrial+fibrillation.">Oxidized Ca2+/calmodulin-dependent protein kinase II triggers atrial fibrillation.</a> Circulation. 128 (16), 1748-1757 (2013).</li><li>Jansen, H. J., 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=Atrial+fibrillation+in+aging+and+frail+mice.">Atrial fibrillation in aging and frail mice.</a> Circulation: Arrhythmia and Electrophysiology. 14 (9), 01077 (2021).</li><li>Luo, T., 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=Characterization+of+atrial+histopathological+and+electrophysiological+changes+in+a+mouse+model+of+aging.">Characterization of atrial histopathological and electrophysiological changes in a mouse model of aging.</a> International Journal of Molecular Medicine. 31 (1), 138-146 (2013).</li><li>McCauley, M. D., 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=Ion+channel+and+structural+remodeling+in+obesity-mediated+atrial+fibrillation.">Ion channel and structural remodeling in obesity-mediated atrial fibrillation.</a> Circulation: Arrhythmia and Electrophysiology. 13 (8), 00896 (2020).</li><li>Kato, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Spectral+analysis+of+heart+rate+variability+during+isoflurane+anesthesia.">Spectral analysis of heart rate variability during isoflurane anesthesia.</a> Anesthesiology. 77 (4), 669-674 (1992).</li><li>Schmeckpeper, J., 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=Abstract+11402:+Targeting+RyR2+to+suppress+ventricular+arrhythmias+and+improve+left+ventricular+function+in+chronic+ischemic+heart+disease.">Abstract 11402: Targeting RyR2 to suppress ventricular arrhythmias and improve left ventricular function in chronic ischemic heart disease.</a> Circulation. 144, 11402 (2021).</li><li>Kim, K., 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=Abstract+B-PO01-017:+RyR2+hyperactivity+promotes+susceptibility+to+ventricular+tachycardia+in+structural+heart+disease.">Abstract B-PO01-017: RyR2 hyperactivity promotes susceptibility to ventricular tachycardia in structural heart disease.</a> Heart Rhythm. 18, 57 (2021).</li></ol>

经食管心房起搏不仅允许在同一动物中进行连续研究，而且其持续时间通常比心内检查短（~20 min），从而最大限度地减少麻醉剂的使用及其对电生理参数的影响。 
针对每个单独的小鼠模型最初优化方法至关重要。衰老增加了正常小鼠的AF诱导性18，19，并且个体遗传模型可能在有限的时间内显示出AF诱导性。每隔一周进行一次试点?...

心房颤动（AF）是多种获得性和遗传性危险因素的最终常见途径。对于研究AF底物病理生理机制的研究，小鼠模型是有利的，因为易于遗传操作，并且通常，它们再现了在人类中观察到的不同临床表型1，2，3的AF敏感性。然而，小鼠很少发生自发性AF4，需要使用激发性心房起搏研究。
可以使用心内5 或经食管6 起搏进行程序性电刺激 （PES） 以评估鼠心房电生理学和 AF 易感性。虽然经食管方法作为生存程序特别有利，但由于许多已发表的实验方案7，8和可能阻碍可重复性的变异源9，其使用变得复杂。此外，有限的报告方案比较使得选择合适的起搏方案具有挑战性。
目前的协议旨在利用系统策略开发模型特异性经食管PES方法来评估鼠AF敏感性，以提高可重复性。重要的是，进行初步试点研究以通过考虑年龄、性别和起搏模式变异性来优化起搏方案，起搏旨在最大限度地减少可能混淆结果的无意副交感神经刺激9.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>27 G ECG electrodes</td>
			<td>ADInstruments</td>
			<td>MLA1204</td>
			<td></td>
		</tr>
		<tr>
			<td>2-F octapolar electrode catheter</td>
			<td>NuMED</td>
			<td>CIBercath</td>
			<td></td>
		</tr>
		<tr>
			<td>Activated carbon canister</td>
			<td>VetEquip</td>
			<td>931401</td>
			<td></td>
		</tr>
		<tr>
			<td>Analysis software</td>
			<td>ADInstruments</td>
			<td>LabChart v8.1.13</td>
			<td></td>
		</tr>
		<tr>
			<td>Biological amplifier</td>
			<td>ADInstruments</td>
			<td>FE231</td>
			<td></td>
		</tr>
		<tr>
			<td>Data acquisition hardware</td>
			<td>ADInstruments</td>
			<td>PowerLab 26T</td>
			<td></td>
		</tr>
		<tr>
			<td>Eye ointment</td>
			<td>MWI Veterinary</td>
			<td>NC1886507</td>
			<td></td>
		</tr>
		<tr>
			<td>Heating pad</td>
			<td>Braintree Scientific</td>
			<td>DPIP</td>
			<td></td>
		</tr>
		<tr>
			<td>Isoflurane</td>
			<td>Piramal</td>
			<td>66794-017-25</td>
			<td></td>
		</tr>
		<tr>
			<td>Stimulator</td>
			<td>Bloom Associates</td>
			<td>DTU-210</td>
			<td></td>
		</tr>
		<tr>
			<td>Stimulus Isolator</td>
			<td>World Precision Instruments</td>
			<td>Model A365</td>
			<td></td>
		</tr>
	</tbody>
</table>

optimization of transesophageal atrial pacing to assess atrial fibrillation susceptibility in mice

心房颤动（AF）的遗传和获得性危险因素的小鼠模型已被证明在研究心房颤动的分子决定因素方面很有价值。程序性电刺激可以使用经食管心房起搏作为生存程序进行，从而能够在同一动物中进行连续测试。然而，存在许多起搏方案，这使得重现性复杂化。本协议旨在提供一种标准化策略来开发特定于模型的实验参数，以提高研究之间的可重复性。进行初步研究以优化所研究特定模型的实验方法，包括研究时的年龄、性别和起搏方案的参数（例如，起搏模式和 AF 易感性的定义）。重要的是，应注意避免高刺激能量，因为这会导致神经节丛刺激，伴有无意的副交感神经激活，表现为起搏期间夸张的房室传导阻滞，并且通常与人为心房颤动感应有关。必须将表现出这种并发症的动物排除在分析之外。

经食管心房起搏研究通过确定SNRT和AVERP以及AF易感性6 来评估SA和AV淋巴结的电生理学特性（图1）。心电图记录可以测量 P 波持续时间、PR 间期、QRS 持续时间和 QT/QTc 间期。在快速心房起搏期间连续记录心电图可以提供以下 AF 脆弱性测量值：研究期间诱导的发作次数、发作的累积和平均持续时间以及持续 AF 发作的次数。起搏期间过度房室传导阻滞的发作可?...

Watch this Scientific Journal Video about Optimization of Transesophageal Atrial Pacing to Assess Atrial Fibrillation Susceptibility in Mice at JoVE.com

Optimization of Transesophageal Atrial Pacing to Assess Atrial Fibrillation Susceptibility in Mice

本协议描述了使用经食管心房起搏评估小鼠心房颤动易感性时实验参数的优化。

优化经食管心房起搏以评估小鼠心房颤动易感性

Mouse models of genetic and acquired risk factors for atrial fibrillation (AF) have proven valuable in investigating the molecular determinants of AF. ...

使用经食管心房起搏的程序性电刺激可以确定小鼠的电生理学特性和对心房颤动的易感性。在这里，我们描述了一种标准化的实验方案开发程序，可以优化重现性。经食管心房起搏方法通常是一种生存过程，因此可以在同一动物中进行连续测试。方案开发的重要组成部分包括初始托盘研究，以优化所研究模型的实验参数，以及识别由起搏诱导的副交感神经刺激引起的心律失常。对于遗传模型，从八周龄开始对小鼠进行每两周一次的心房起搏，以确定AF易感性的最佳时期，研究两性，因为一个人可能不会发展出AF表型。对于获得的模型，在小鼠达到身体成熟后进行起搏。虽然通常在 12 周时达到成熟，但根据需要调整实验模型的年龄。执行突发和减搏以确定最佳起搏模式。将每个程序分开至少 24 小时。使用 AF 敏感性的多种定义分析数据。使用优化的模型特定参数和AF敏感性的定义进行其他小鼠的后续研究。踏板反射丧失后，将麻醉小鼠仰卧放在加热垫上，加热垫旨在将体温保持在约 37 摄氏度，后肢贴在垫表面上。通过将连接到生物放大器和数据采集硬件的 27 个规格 ECG 针电极皮下放置到前肢中，并在后肢中放置接地电极，获得表面心电导联。将连接到刺激器和刺激隔离器的双法式八极电极导管插入食道。插入到近似从口腔到剑突软骨上方的距离的深度，颈部伸展。使用分析软件连续记录心电图导联开始数据采集。将导管正确放置在食道内以启用捕获。为此，在略短于正弦周期长度的周期长度下施加脉冲宽度为 2 毫秒的 1.5 毫安刺激。小心放置导管，直到获得一致的心房捕获。要确定心房舒张期捕获阈值，请在用于心房捕获的周期长度处以 1.5 毫安的脉冲宽度 2 毫秒启动起搏。将刺激振幅减小 0.05 毫安增量，直到心房捕获丢失，随后增加直至捕获。获得尽可能低的起搏阈值，以尽量减少无意起搏引起的副交感神经刺激。将刺激幅度调整到阈值的两倍。测量心房快速起搏前的电生理学参数，包括窦房结恢复时间、Wenckebach 周期长度和房室有效不应期以进行心房诱导。使用不同周期长度的突发起搏或初始研究确定的降低起搏，以两毫秒的脉冲宽度以阈值的两倍执行起搏。对于突发配速，以 50 毫秒的初始周期长度配速 15 秒，然后以 40、30、25、20 和 15 毫秒的周期长度配速。对于降低配速，执行三到五列火车，并以 40 毫秒的周期长度配速，每两秒减少 2 毫秒，直到在 20 毫秒处终止。在最后一次起搏训练后 30 秒的窦性心律或 10 分钟的 AF 发作后终止手术，以先到者为准。轻轻取出导管和心电图电极。在串行测试的情况下，请至少等待 24 小时，然后再重复起搏过程。心电图记录可以测量 P 波持续时间、PR 间期、QRS 持续时间以及 QT 和 QTC 间期。在快速心房起搏期间连续记录心电图提供了研究期间诱导的每次 AF 发作的记录。根据这些数据，记录了发作的累积和平均持续时间以及持续AF发作的次数。起搏期间房室传导阻滞过多的发作表明起搏诱导的副交感神经刺激的时期，表明任何相关的心房颤动都是这种现象的伪影，而不是模型本身的病理生理学。这是由于位于左心房后部的神经节丛的无意刺激引起的，导致副交感神经激活。这种类型的心律失常诱导增加了对照小鼠中AF的发生率，并在实验组中导致更大的心律失常变异性。鉴于这些污染特征，在这些条件下经历AF的动物应从分析中排除。当使用该实验策略时，初步试点研究和识别导致无意中AF诱导的副交感神经刺激对于方案优化和研究可重复性尤为重要。

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Methods to Assess Subcellular Compartments of Muscle in C. elegans

方法来评估肌肉的亚细胞中<em&gt;温度。线虫</em

Muscle is a dynamic tissue that responds to changes in nutrition, exercise, and disease state. The loss of muscle mass and function with disease and age ...

Voltage and Calcium Dual Channel Optical Mapping of Cultured HL-1 Atrial Myocyte Monolayer

培养HL-1心房肌细胞单层的电压和钙双通道光映射

Optical mapping has proven to be a valuable technique to detect cardiac electrical activity on both intact ex vivo hearts and in cultured myocyte ...

Gap Junctional Intercellular Communication: A Functional Biomarker to Assess Adverse Effects of Toxicants and Toxins, and Health Benefits of Natural Products

间隙连接通讯：天然产物的生物标志物的功能，以评估毒物和毒素的不利影响，以及健康的益处

This protocol describes a scalpel loading-fluorescent dye transfer (SL-DT) technique that measures intercellular communication through gap junction ...

High-resolution Optical Mapping of the Mouse Sino-atrial Node

鼠标窦房结的高分辨率光学测绘

Sino-atrial node (SAN) dysfunctions and associated complications constitute important causes of morbidity in patients with cardiac diseases. The ...

A Method to Assess Bacteriocin Effects on the Gut Microbiota of Mice

评估细菌素对小鼠肠道微生物的影响的方法

Very intriguing questions arise with our advancing knowledge on gut microbiota composition and the relationship with health, particularly relating to the ...

Simultaneous Isolation and Culture of Atrial Myocytes, Ventricular Myocytes, and Non-Myocytes from an Adult Mouse Heart

来自成人鼠心脏的心外肌细胞、心室肌细胞和非肌细胞的同步隔离和文化

The isolation and culturing of cardiac myocytes from mice has been essential for furthering the understanding of cardiac physiology and pathophysiology. ...