This work was supported by the Huffington Center on Aging at Baylor College of Medicine, Geriatrics, CVS DeBakey Heart Center at Houston Methodist Hospital and BCM, NIH RO-1 HL 13870 to ML Entman, as well as a Career Development Award # IK2BX002410 from the United States Department of Veterans Affairs (LMP), Biomedical Laboratory Research and Development Program.
We are immensely grateful to Mark Entman M.D. for sharing his wisdom and supporting this work.

所有的动物都按照健康指南全国学院的照顾和实验动物使用的贝勒医学院的指导方针照顾。 1.建立<ol><li>打开超声心动图系统。从下拉菜单中选择显示"心脏测量"启动软件并点击"初始化"。 </li><li>首先在屏幕的上半部分的"新建"按钮，一项新的研究点击进入学习鼠标的人口统计信息。例如鼠标ID，性别，出生日期，以及操作人员做研究的名字。 </li><li>通过点击"TEMP ON / OFF"按钮打开温控EKG板。将其设置为使用向上和向下箭头的39℃的温度。 </li><li>要开始设立麻醉室，首先确保氧气和异氟烷水平在各输送系统的合适。 </li><l我&gt;将鼠标在麻醉室，并与异氟醚和氧气，直到卧位诱导，然后将其传送到口罩。确认鼠标，已经正确捏它的爪子和检查缺乏运动麻醉。保持在可避免的正爪缩回响应可能的最低水平的异氟烷。 </li><li>将鼠标在温度控制EKG板仰卧位和磁带的每个爪子板上的相应传感器元件。或者，使用传感器霜来改善信号。 </li><li>鼠标的眼睛应用人工泪液眼软膏的润滑剂，以防止干燥。 </li><li>一旦鼠标贴在EKG板，观察心电图在显示屏的下部的跟踪。监测在整个研究的心脏速率，以确保最佳结果。 400次的心脏速率/分钟是理想的。低于此范围的心脏速率可能暗示异氟醚的效果。 </li><li>确保鼠标不断地接受和氧气通过面罩异氟醚。 </li><li>使用电动剃须刀，从鼠标的前胸部及上腹部区剃除身上的皮毛一体。使用脱毛霜如果必要，以进一步确保皮毛去除。 </li><li>把传感器凝胶大方量在鼠标的胸部。 </li></ol> 2.图像采集<ol><li> 左心房测量 <ol><li>找到EKG板下方的手柄。使用手柄调整可动EKG板，其中将小鼠放置的角度。沿X轴和沿Y轴为0°角45°角 - 由心电图板调整到30开始。作者将其称为"仰卧"位置。请对所有左心房测量获取这一立场。 </li><li>放置在超声心动图可动臂夹紧的25MHz的扫描头在长轴图。确保选择的超声心动图系统的心脏测量滤波器和二维B模式。 </li><li>注意，回声心动描记集具有可以顺时针或逆时针移动的X和Y轴调节扭矩。旋转它们在主动脉流出道的水平，以获得心脏的长轴，并获得5秒动画拍摄。通过按下键盘的超声心动图系统的"电影商店"按钮获得的扫描。使用此计算LA的优越劣测量。 </li><li>电影收购完成后，按键盘的超声心动图系统的"M-MODE"键切换超声心动图，以M-模式。 </li><li>观察矢状图和光标。将光标，以便它包含整个洛杉矶在主动脉流出道的电平。获得这一新观点的3张照片。通过按下键盘的超声心动图系统的"帧存储"按钮获得的扫描。使用此计算LA的前后测量。 </li><li>通过按下"B型"返回B型，但<html>吨键盘超声系统上。 </li><li>旋转集顺时针90°的可动臂夹紧。这将在短轴视图设置扫描头。 </li><li>使用Y轴扭矩发现关节内隔。确保想象中庭的墙壁上。获得5秒动画拍摄。通过按下键盘的超声心动图系统的"电影商店"按钮获得的扫描。使用此计算LA的内 - 外的测量。 </li></ol></li></ol><img alt="图1" src="/files/ftp_upload/55214/55214fig1.jpg" /> 图1:"仰卧"位置。在"仰卧"位置鼠标的表示。约30 - 沿X轴45°角度并沿Y轴为0°角。这个位置是用来收购所有左心房测量。T ="_空白"&gt;点击此处查看该图的放大版本。 <ol start="2"><li> 大血管测量 <ol><li>通过为30 MHz的扫描头交换25 MHz的扫描头继续。除去从可动臂夹紧的25MHz的扫描头，并将30MHz的扫描头代替。确保选择的超声心动图系统的腹部测量滤波器和二维B模式。 </li><li>找到EKG板下方的手柄。使用手柄来调整移动EKG板，其中鼠标所在的角度。由心电图板调整到沿X轴和沿Y轴60°角度的5°角开始。使用此podition收购的主动脉和肺动脉的测量。称此为"起立"的位置。 </li><li>在主动脉流出道的水平，超声心动图组使用X和Y轴调节扭矩，让心脏的长轴，并获得5秒动画拍摄。收购日e。通过按键盘的超声心动图系统的"电影商店"按钮进行扫描。用这个来计算主动脉内径。 </li><li>电影收购完成后，集旋转顺时针90°的动臂夹紧。这将在短轴视图设置扫描头。 </li><li>使用Y轴扭矩定位在其分叉水平的肺动脉，并获得5秒动画拍摄。通过按下键盘的超声心动图系统的"电影商店"按钮获得的扫描。用这个来计算肺动脉内径。 注:研究人员可以利用多普勒成像作为一种工具，以确保肺动脉正确识别。如果需要的话，一旦完成步骤2.2.5按键盘的超声心动图系统获得多普勒成像对"私服"按钮。如果肺特征描记波被视为，研究者可以放心正确的位置被确定。 </li></ol></li></ol<html> &gt; <img alt="图2" src="/files/ftp_upload/55214/55214fig2.jpg" /> 图2:"起立"的位置。在"起立"位置鼠标的表示。大约5°角沿X轴和沿Y轴60°角。这个位置是用来采集主动脉和肺动脉直径。 <a href="https://ecsource-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/55214/55214fig2large.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/55214/55214fig3.jpg" /> 图3:肺动脉多普勒。特点肺动脉多普勒成像。研究人员可以利用多普勒成像作为一种工具，以确保肺动脉正确识别。空白"&gt;点击此处查看该图的放大版本。 3.程序后恢复动物<ol><li>为了安全地删除该EKG板鼠标，请从鼠标的胸部凝胶从四肢取出磁带。 </li><li>关掉麻醉和氧气输送系统。返回鼠标笼子里，直到它已经恢复了足够的意识，以保持胸骨斜卧不要离开鼠标无人值守。 </li></ol> 4.图像分析<ol><li> 图像传输 <ol><li>使用USB闪存驱动器中的超声心动图系统收集到的图像传输到计算机。将USB闪存驱动器插入的超声心动图系统的USB端口。 </li><li>选择学习或研究，通过点击研究姓名（或名称）来传输。 </li><li>点击"复制到"位于屏幕的上部按钮。显示文件和目录将应用程式的新窗口耳。选择与刚插入的USB闪存驱动器的目录，然后单击"确定"。 </li><li>拆下超声心动图系统中的USB闪存驱动器，然后将其插入计算机的USB端口用于分析。 </li><li>启动超声心动图软件。点击"复制自"位于屏幕的上部按钮。观察显示文件和目录的新窗口。选择研究，从对应于刚插入的USB闪存驱动器的目录进行复制，然后单击"确定"。 </li><li>复制文件后，观察他们在研究名单。要开始分析其名称的研究，双击并选择操作分析的名称。 </li></ol></li><li> 左心房分析 注:以下所有心房测量使用EKG为指导，以选择P波为那里的研究人员希望测量LA点。这对应于前心房收缩时第ËLA是其最大容量。 <ol><li>选择对应于左心房（步骤2.1.3）的高级劣尺寸的影片剪辑。确定使用心电图作为引导心房预收缩时间。这样的空腔在它的最大容量进行测定。 </li><li>从屏幕顶部的工具栏上，单击"工具"，然后选择"测量"。将出现一个新的窗口。点击"线性距离测量工具"图标（它对应的第一个图标在窗口的左上角）。测量其最大直径腔。 </li><li>选择对应于左心房（步骤2.1.5）的前后尺寸图像。确定使用心电图作为引导心房预收缩时间。这样的空腔在它的最大容量进行测定。为了衡量重复步骤4.2.2。 </li><li>选择对应于左心房（步骤2.1.8）的内 - 外尺寸图像。确定预心房承包商使用心电图作为指导和灰的时间。这样的空腔在它的最大容量进行测定。测量从房间隔的相对壁的空腔中的对角的方式。为了衡量重复步骤4.2.2。 注:重复步骤4.2.1直通4.2.4至少3次，并获得平均值，以减少错误。 </li></ol></li><li> 大血管分析 注:以下所有大血管测量使用EKG为指导后，QRS波群为那里的研究人员希望测量点选择时间。这对应于大血管是在其最大容量的点。 <ol><li>选择对应于主动脉（步骤2.2.3）的短片。确定使用心电图QRS波之后的一个点。使用血管壁作为基准测量在90°角的直径。为了衡量重复步骤4.2.2 </li><li>选择对应于肺动脉（步骤2.2.5）的短片。决定在Q后点RS使用EKG。使用血管壁作为基准测量在90°角的直径。选择肺动脉分叉以上的平面。为了衡量重复步骤4.2.2。 </li></ol></li></ol>

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The authors have nothing to disclose.

有成功测量LAV，主动脉和PA直径的三个关键步骤。在设定了它完全删除在胸前的毛皮是非常重要的;如果不这样做会导致图像质量的干扰。在图像采集步骤，重要的是获得具有其相应的探针和过滤器各图像每秒帧从探头变化来探测:即25兆赫和心脏筛选为LAV的评估和30兆赫和评估腹部过滤的主动脉和PA的直径。研究者有时由图像获取差如果鼠标具有的脂肪量很大，或者如果一个肋使得难以得到良好的窗口来限制。在图像分析步骤中的代表​​结果部分所述，以测量它们的最大容量的腔是很重要的。使用EKG为指导。 超声心动图已知是依赖于操作者。这种技术需要熟悉与心脏解剖和超声心动图的窗户最佳图像采集，以及减少学习时间。熟悉变得相关，以防止长时间使用异氟醚的衍生如心脏速率和血流动力学妥协减慢并发症。为了防止这种情况，该动物的密切监测必须通过保持整个研究的心脏速率高于400次/分得到保证。 该技术提供了相对于其他可用的技术来评估在小鼠中的心脏功能的许多优点。相比侵入性导尿它是允许连续测量一个非侵入性，非终端技术;相比MRI，它是便宜和更快8。通过二维超声心动图测量LAV和PA直径这一新技术打开大门，监控措施和心肺科学的进一步研究。

目前的方法教调查员如何衡量和使用LAV，主动脉直径和PA直径二维超声心动图小鼠下异氟醚。心脏衰竭与老年人射血分数（HFPEF）影响的那些80年高达10％及以上1和结果显著发病率和死亡率。显著的死亡率也由肺动脉高压（PH），一个阴险的疾病过程有类似症状HFPEF，其中肺动脉压力升高导致劳力性呼吸困难，进行正确的心脏衰竭，并经常提出的死亡发生。 2既HFPEF和PH值的日益流行表示用于开发了一种方法，允许在非侵入性，非终端的方法准确的评价，并在小鼠模型干预监测。 通过心室艺术改变老化导致舒张功能恶化erial硬化，血管功能障碍，炎症3，受损的钙调节4，降低β肾上腺素能响应性和身体功能失调产生积极的减缓放松和增加被动刚度。随着时间的推移这将导致增加的LV充盈压和洛杉矶的代偿性肿大。 五 虽然其他病因，如瓣膜功能不全（二尖瓣关闭不全或狭窄）和浸润工艺导致在LA 1压力和容积升高，欧洲心脏病学会支持添加LA大小为LV功能的无创性反映。 6 洛杉矶量和舒张功能的微创措施之间的相关性进行了研究，小鼠。在功能差异相关洛杉矶量确定侵入年龄组为14位和31个月之久的米冰。在14个月大的老鼠，LA体积舒张功能的三个标准侵入措施相关-dP / DTMIN（R 2 = 0.5，P &lt;0.05），头（松弛时间常数（R 2 =。6， p &lt;0.05），左室舒张末压（R 2 = 0.25，p &lt;0.05）; 31个月大的老鼠，洛杉矶量和-dP / DTMIN（R 2 = 0.92，p之间的关系&lt; 0.05），左室舒张末压（R 2 = 0.61，p &lt;0.05），但明显与头的关系是不太清楚。因此，洛杉矶量不仅跨组，但年龄组中舒张功能障碍增加。7 在使用导管技术来评估心脏功能，虽然严格而可靠的，被限制的鼠模型的心脏功能的研究由于其与反复评估不相容性。 8替代侵入的措施，如MRI和三维超声心动图也可以是比2D超声心动图技术更准确，但它们比较昂贵; 2D超声心动图被认为足以洛杉矶量评估。 9，10 对LA体积和PA直径与超声心动图评估允许产生在肺动脉阻力主要增加导致与在洛杉矶的压力或LAV从那些没有变化的增加的PA直径，其中肺动脉和左心房都放大作为模型之间的区别对心脏的左侧充盈压升高的结果。这种方法是采取斯卡利亚等。谁发现，在人，超声心动图肺动脉左心房比（ePLAR）是给患者预先肺毛细血管高血压和capillaryhypertension柱之间精确区分的参数。 11

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			<td height="42" style="height:42px;width:203px;">Vevo 770 high-resolution in vivo micro-imaging system</td>
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			<td style="width:267px;">Echocardiographic Equipment</td>
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			<td height="84" style="height:84px;width:203px;">707B RMV (Real time MicroVisualization) 30 MHz Scanhead with encapsulated transducer&#160;</td>
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			<td>Real Time Microvisualization Scanhead</td>
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			<td height="84" style="height:84px;width:203px;">710B RMV (Real time MicroVisualization) 25 MHz Scanhead with encapsulated transducer&#160;</td>
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			<td style="width:119px;">710B-159</td>
			<td>Real Time Microvisualization Scanhead</td>
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			<td height="63" style="height:63px;width:203px;">Vevo integrated rail system including physiological monitoring unit&#160;</td>
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			<td height="21" style="height:21px;width:203px;">Standard Gauze pads&#160;&#160;</td>
			<td style="width:85px;">McKeeson</td>
			<td style="width:119px;">General supply</td>
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			<td height="21" style="height:21px;width:203px;">Tape</td>
			<td style="width:85px;">Durapore</td>
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			<td height="21" style="height:21px;width:203px;">Nose cone&#160;</td>
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			<td>For anesthesia delivery</td>
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murine echocardiography of left atrium, aorta, and pulmonary artery

本方法教导研究者如何测量并使用LAV通过超声心动图中左心室舒张压慢性升高的替代，以及获得小鼠主动脉和PA直径的测量。 比的年龄10天老年小鼠中可以使用本技术进行分析。该技术是由3个主要步骤:设置，图像采集和图像分析。的设立步骤包括得到含有1％异氟烷麻醉小鼠，剃须它，并在仰卧位置到加热的EKG板，其中所述图像采集将于胶带它的。图像获取步骤包括学习以识别心脏结构和获得所有与它的相应探针和轴所需的图像，以便能够计算体积和直径。图像分析步骤包括在计算机软件的帮助下测定以前获得的图像。

对LA体积和PA直径与超声心动图评估允许其中增加PA的直径没有变化的LAV从那些肺动脉和左心房扩大都在充盈压升高的两个结果增加肺动脉阻力结果模型之间的区别左侧的心脏。显示在两个病理生理学差异图像示于图4。洛杉矶和PA的图像从年轻，缺氧引起肺动脉高压和年龄有关的左心室舒张功能障碍。肺动脉高压模型的发展离不开左心房容积超过4周周期的变化增加了PA的直径。相比较而言，老的动物发展在两个其肺动脉直径和左心房容积暗示的压力向后传播驱动此组中的PA直径的增加的增加。 图4:LA上-下和PA直径在普通鼠标，肺动脉高压模型小鼠和老龄小鼠与年龄相关的左室舒张功能不全。图像（A）和（B）分别有15周龄正常小鼠显示LA优越劣和PA的直径。图像（C）和（D）分别有15周龄肺动脉高压模型小鼠表现出LA优越劣和PA的直径。肺动脉高血压诱导通过暴露于10％的低氧四周并通过右心脏导管研究PA的收缩压增加75％。请注意在PA直径的差异。图像（E）和（F）分别为一名21个月大的老鼠与高低压端舒张压显示LA优越劣和PA的直径。填充压力瓦特作为老老鼠增加了近300％。请注意，无论是洛杉矶和PA直径较大相比15周龄的小鼠。规模毫米单位。标记"Δ"表示在所得到所示的图像的心电图的点。 LA SI:左心房上 - 下，PA:肺动脉。 <a href="https://ecsource-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/55214/55214fig4large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> 为了确保精确度的结果是必要的测量是在心动周期在它们的适当的定时作出。当测量LAV，研究人员希望在其最大的三个措施的能力来衡量的心房，这意味着心房收缩之前。如果左心耳是在上 - 下视图的任何小鼠中观察，作者建议做测量时，忽略它。当测量大血管研究员婉TS来衡量他们在其最大容量，QRS后或之后心室泵有血液进入血管的意义。 图5至图 9示出对应于相同的动物，在不同时间点上的心电图测定图像示出了正确的和不正确的测量之间的差异。 研究者现在可以使用扁长椭圆9的公式计算LAV，10如下面的例子: （4 *π*上 - 下维*前 - 后尺寸*内 - 外尺寸）/（3 * 2 * 2 * 2） 在图5中 ，一个"正确"的测量是使用心电图为导向，以确保研究者在空腔的最大容量测量所获得的。在这种情况下吨他心房是其最大容量时，只需在P波测量。 （A，C）。一个"不正确"的测量是在比P波以外的任何点获得。它往往给更小的尺寸为左心房合同或填充。 （B，D）。在图6中 ，一个"正确"的测量是使用心电图为导向，以确保研究者在空腔的最大容量测量所获得的。在这种情况下，当只是在P波测量的心房是在其最大容量（LA（1））。一个"不正确"的测量是在比P波以外的任何点获得。它往往给更小的尺寸为左心房合同或填充。 （LA（2））。在图7中 ，一个"正确"的测量是使用心电图为导向，以确保研究者在空腔的最大容量测量所获得的。当只有在P波测量的心房是在其最大容量。 （A，C）。一个"不正确"的测量是获得编在比P波以外的任何点。 （B，D）。 *笔者认为，电影是确定LA的限制非常有帮助。研究人员应该能够看到整个心动周期洛杉矶之举。在图8中 ，一个"正确"的测量是使用心电图为导向，以确保研究人员在容器的最大容量测量所获得的。在这种情况下，主动脉是在刚刚QRS复合后的最大容量。 （A，C）。一个"不正确"的测量是，在不仅仅是QRS波群之后，其他任何时候获得的。它往往给更小的尺寸为主动脉合同或填充。 （B，D）。在图9中 ，一个"正确"的测量是使用心电图为导向，以确保研究人员在容器的最大容量测量所获得的。在这种情况下，肺动脉是在刚刚QRS复合后的最大容量。 （A，C）。 ，在的所有P获得了"不正确"的测量oint不仅仅是QRS波群后，其他。它往往给更小的尺寸为肺动脉合同或填充。 （B，D）。 <img alt="图5" src="/files/ftp_upload/55214/55214fig5.jpg" /> 图5:LA上-下测量。图像（A）和（B）对应于相同的鼠标标本。规模毫米单位。图像（C）对应于图像心电图（A）的右下角片段。不按比例。图像（D）的对应于图像（B）的EKG的右下片段。不按比例。注意图像中的标记"Δ"（C）和（D）。标记"Δ"表示在所得到所示的图像的心电图的点。 <a href="https://ecsource-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/55214/55214fig5large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> <img alt="图6" src="/files/ftp_upload/55214/55214fig6.jpg" /> 图6:LA前后测量。左心房的M型视图。该图像对应于左心房的M型视图，以获得前后测量。 <a href="https://ecsource-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/55214/55214fig6large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> <img alt="图7" src="/files/ftp_upload/55214/55214fig7.jpg" /> 图7:LA内-外测量。图片 （A）和（B）中对应于相同的鼠标标本。规模毫米单位。 （C）对应的心电图（A）的片段。不按比例。 （D）的对应的心电图（B）的一个片段。不规模。注意图像中的标记"Δ"（C）和（D）。标记"Δ"表示在所得到所示的图像的心电图的点。 LV:左室，RA:右心房，LA:左心房。 <a href="https://ecsource-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/55214/55214fig7large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> <img alt="图8" src="/files/ftp_upload/55214/55214fig8.jpg" /> 图8:主动脉测量。图片 （A）和（B）中对应于相同的鼠标标本。规模毫米单位。 （C）对应的心电图（A）的片段。不按比例。 （D）的对应的心电图（B）的一个片段。不按比例。注意图像中的标记"Δ"（C）和（D）。标记"Δ"表示在心电图在该点得到所示的图像。 <a href="https://ecsource-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/55214/55214fig8large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> <img alt="图9" src="/files/ftp_upload/55214/55214fig9.jpg" /> 图9:肺动脉测量。图像（A）和（B）对应于相同的鼠标标本。规模毫米单位。 （C）对应的心电图（A）的片段。不按比例。 （D）的对应的心电图（B）的一个片段。不按比例。注意图像中的标记"Δ"（C）和（D）。标记"Δ"表示在所得到所示的图像的心电图的点。 <a href="https://ecsource-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/55214/55214fig9large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> 为了演示的重现和该技术的一致性，作者提供从他们自己的工作衍生的两个例子。第一个提出了一组19个月大的老鼠，一个星期后进行了基线超声心动图和后续的心动。作者收集左心房和大血管的测量如先前描述的和所获得的结果如下。左侧图显示采用扁长的椭圆公式9，10计算每个鼠标的左心房卷。右边图表显示每个老鼠的肺动脉内径。数据显示的只有百分之几变动。 <img alt="图10" src="/files/ftp_upload/55214/55214fig10.jpg" /> 图10:左房容积和肺动脉内径上的两个连续的屁股<html> essments。 （A）显示在六个19个月的小鼠立方毫米左心房卷2后续评估。 （B）显示了相同的19个月的小鼠组毫米肺动脉内径。 <a href="https://ecsource-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/55214/55214fig10large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> 作为第二个例子，本作者由两个不同的研究人员在同一天获得相同21个月老小鼠的两个图像。前两个面板对应于由研究员＃1和第二两个面板获取的图像对应于由研究员＃2获取的图像。请注意，在"时间增益补偿"的差别不影响测量。这个例子表明选准点中的QRS，同时测量空腔大小，如先前所描述的极大重要性。 jove_content"FO: - together.within页保留="1"&gt; <img alt="图11" src="/files/ftp_upload/55214/55214fig11.jpg" /> 图11:LA上-下和PA直径由两个不同的研究人员获得了相同的19个月的鼠标。 （A）显示左心房上-下直径。 （B）表示同一小鼠的PA的直径。这两个图像是由在同一天同一研究者获得的。 （C）表示左心房上-下直径。 （D）表示相同的鼠标的PA的直径。这两个图像是由在同一天的第二研究者获得的。规模毫米单位。标记"Δ"表示在所得到所示的图像的心电图的点。 LA SI:左心房上 - 下。 <a href="https://ecsource-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/55214/55214fig11large.jpg" target="_blank">请点击此处查看该图的放大版本。</a>

Watch this Scientific Journal Video about Murine Echocardiography of Left Atrium, Aorta, and Pulmonary Artery at JoVE.com

Murine Echocardiography of Left Atrium, Aorta, and Pulmonary Artery

以下方案描述了用于获得左心房容积（LAV），主动脉（AO）的直径，以及肺动脉（PA）的直径在小鼠心脏超声图像的获取和分析的方法。这种技术是一种非侵入性的，非终端程序，允许心肺功能的评估。

左心房，主动脉和肺动脉的超声心动图小鼠

The present methodology teaches the investigator how to measure and use the LAV as a surrogate of chronic elevations in Left Ventricular diastolic ...

murine-echocardiography-of-left-atrium-aorta-and-pulmonary-artery

Nanyang Technological University

Research

JoVE Journal

Medicine

14.2K 次观看.  Baylor College of Medicine. 以下方案描述了用于获得左心房容积（LAV），主动脉（AO）的直径，以及肺动脉（PA）的直径在小鼠心脏超声图像的获取和分析的方法。这种技术是一种非侵入性的，非终端程序，允许心肺功能的评估。 

视频: Murine Echocardiography of Left Atrium, Aorta, and Pulmonary Artery

Murine Echocardiography and Ultrasound Imaging

Rodent models of cardiac pathophysiology represent a valuable research tool to investigate mechanism of disease as well as test new therapeutics.1 Echocardiography provides a powerful, non-invasive tool to serially assess cardiac morphometry and function in a living animal.2 However, using this technique on mice poses unique challenges owing to the small size and rapid heart rate of these animals.3 Until recently, few ultrasound systems were capable of performing quality echocardiography on mice, and those generally lacked the image resolution and frame rate necessary to obtain truly quantitative measurements. Newly released systems such as the VisualSonics Vevo2100 provide new tools for researchers to carefully and non-invasively investigate cardiac function in mice. This system generates high resolution images and provides analysis capabilities similar to those used with human patients. Although color Doppler has been available for over 30 years in humans, this valuable technology has only recently been possible in rodent ultrasound.4,5 Color Doppler has broad applications for echocardiography, including the ability to quickly assess flow directionality in vessels and through valves, and to rapidly identify valve regurgitation. Strain analysis is a critical advance that is utilized to quantitatively measure regional myocardial function.6 This technique has the potential to detect changes in pathology, or resolution of pathology, earlier than conventional techniques. Coupled with the addition of three-dimensional image reconstruction, volumetric assessment of whole-organs is possible, including visualization and assessment of cardiac and vascular structures. Murine-compatible contrast imaging can also allow for volumetric measurements and tissue perfusion assessment.

This video demonstrates use of a rail-mounted high-frequency ultrasound probe to perform echocardiography on an anesthetized mouse. The methods describe both conventional two-dimensional and M-mode measurements of cardiac function in addition to newer, more powerful tools such as color Doppler, strain analysis, as well as general and targeted contrast imaging.

小鼠超声心动图和超声成像

Rodent models of cardiac pathophysiology represent a valuable research tool to investigate mechanism of disease as well as test new therapeutics.1  ...

科研

医学

Murine Fetal Echocardiography

Transgenic mice displaying abnormalities in cardiac development and function represent a powerful tool for the understanding the molecular mechanisms underlying both normal cardiovascular function and the pathophysiological basis of human cardiovascular disease. Fetal and perinatal death is a common feature when studying genetic alterations affecting cardiac development 1-3. In order to study the role of genetic or pharmacologic alterations in the early development of cardiac function, ultrasound imaging of the live fetus has become an important tool for early recognition of abnormalities and longitudinal follow-up. Noninvasive ultrasound imaging is an ideal method for detecting and studying congenital malformations and the impact on cardiac function prior to death 4. It allows early recognition of abnormalities in the living fetus and the progression of disease can be followed in utero with longitudinal studies 5,6. Until recently, imaging of fetal mouse hearts frequently involved invasive methods. The fetus had to be sacrificed to perform magnetic resonance microscopy and electron microscopy or surgically delivered for transillumination microscopy. An application of high-frequency probes with conventional 2-D and pulsed-wave Doppler imaging has been shown to provide measurements of cardiac contraction and heart rates during embryonic development with databases of normal developmental changes now available 6-10. M-mode imaging further provides important functional data, although, the proper imaging planes are often difficult to obtain. High-frequency ultrasound imaging of the fetus has improved 2-D resolution and can provide excellent information on the early development of cardiac structures 11.

Fetal and perinatal death is a common feature when studying genetic alterations affecting cardiac development. High-frequency ultrasound imaging has improved 2-D resolution and can provide excellent information on early cardiac development and is an ideal method to detect the impact on cardiac structure and function prior to death.

小鼠胎儿超声心动图

Transgenic mice displaying abnormalities in  cardiac development and function represent a powerful tool for the understanding  the molecular mechanisms ...

Assessment of Right Ventricular Structure and Function in Mouse Model of Pulmonary Artery Constriction by Transthoracic Echocardiography

Emerging clinical data support the notion that RV dysfunction is critical to the pathogenesis of cardiovascular disease and heart failure1-3. Moreover, the RV is significantly affected in pulmonary diseases such as pulmonary artery hypertension (PAH). In addition, the RV is remarkably sensitive to cardiac pathologies, including left ventricular (LV) dysfunction, valvular disease or RV infarction4. To understand the role of RV in the pathogenesis of cardiac diseases, a reliable and noninvasive method to access the RV structurally and functionally is essential.
A noninvasive trans-thoracic echocardiography (TTE) based methodology was established and validated for monitoring dynamic changes in RV structure and function in adult mice. To impose RV stress, we employed a surgical model of pulmonary artery constriction (PAC) and measured the RV response over a 7-day period using a high-frequency ultrasound microimaging system. Sham operated mice were used as controls. Images were acquired in lightly anesthetized mice at baseline (before surgery), day 0 (immediately post-surgery), day 3, and day 7 (post-surgery). Data was analyzed offline using software.
Several acoustic windows (B, M, and Color Doppler modes), which can be consistently obtained in mice, allowed for reliable and reproducible measurement of RV structure (including RV wall thickness, end-diastolic&#160;and end-systolic dimensions), and function (fractional area change, fractional shortening, PA peak velocity, and peak pressure gradient) in normal mice and following PAC.
Using this method, the pressure-gradient resulting from PAC was accurately measured in real-time using Color Doppler mode and was comparable to direct pressure measurements performed with a Millar high-fidelity microtip catheter. Taken together, these data demonstrate that RV measurements obtained from various complimentary views using echocardiography are reliable, reproducible and can provide insights regarding RV structure and function. This method will enable a better understanding of the role of RV cardiac dysfunction.

Right ventricle (RV) dysfunction is critical to the pathogenesis of cardiovascular disease, yet limited methodologies are available for its evaluation. Recent advances in ultrasound imaging provide a noninvasive and accurate option for longitudinal RV study. Herein, we detail a step-by-step echocardiographic method using a murine model of RV pressure overload.

右心室结构和功能的肺动脉缩窄经胸超声小鼠模型的评估

Emerging clinical data support the notion that RV dysfunction is critical to the pathogenesis of cardiovascular disease and heart failure1-3. Moreover, ...

A Murine Model of Myocardial Ischemia-reperfusion Injury through Ligation of the Left Anterior Descending Artery

Acute or chronic myocardial infarction (MI) are cardiovascular events resulting in high morbidity and mortality. Establishing the pathological mechanisms at work during MI and developing effective therapeutic approaches requires methodology to reproducibly simulate the clinical incidence and reflect the pathophysiological changes associated with MI. Here, we describe a surgical method to induce MI in mouse models that can be used for short-term ischemia-reperfusion (I/R) injury as well as permanent ligation. The major advantage of this method is to facilitate location of the left anterior descending artery (LAD) to allow for accurate ligation of this artery to induce ischemia in the left ventricle of the mouse heart. Accurate positioning of the ligature on the LAD increases reproducibility of infarct size and thus produces more reliable results. Greater precision in placement of the ligature will improve the standard surgical approaches to simulate MI in mice, thus reducing the number of experimental animals necessary for statistically relevant studies and improving our understanding of the mechanisms producing cardiac dysfunction following MI. This mouse model of MI is also useful for the preclinical testing of treatments targeting myocardial damage following MI.

We introduce a surgical method to induce experimental ischemia/reperfusion (I/R) injury to simulate myocardial infarction (MI) in mouse models that allows for more clarity in positioning of the ligation on the left anterior descending artery (LAD) to increase the reproducibility of MI experiments in mice.

心肌缺血再灌注损伤的通过左前降支结扎小鼠模型

Acute or chronic myocardial infarction (MI) are cardiovascular events resulting in high morbidity and mortality. Establishing the pathological mechanisms ...

Increasing Pulmonary Artery Pulsatile Flow Improves Hypoxic Pulmonary Hypertension in Piglets

Pulmonary arterial hypertension (PAH) is a disease affecting distal pulmonary arteries (PA). These arteries are deformed, leading to right ventricular failure. Current treatments are limited. Physiologically, pulsatile blood flow is detrimental to the vasculature. In response to sustained pulsatile stress, vessels release nitric oxide (NO) to induce vasodilation for self-protection. Based on this observation, this study developed a protocol to assess whether an artificial pulmonary pulsatile blood flow could induce an NO-dependent decrease in pulmonary artery pressure. One group of piglets was exposed to chronic hypoxia for 3 weeks and compared to a control group of piglets. Once a week, the piglets underwent echocardiography to assess PAH severity. At the end of hypoxia exposure, the piglets were subjected to a pulsatile protocol using a pulsatile catheter. After being anesthetized and prepared for surgery, the jugular vein of the piglet was isolated and the catheter was introduced through the right atrium, the right ventricle and the pulmonary artery, under radioscopic control. Pulmonary artery pressure (PAP) was measured before (T0), immediately after (T1) and 30 min after (T2) the pulsatile protocol. It was demonstrated that this pulsatile protocol is a safe and efficient method of inducing a significant reduction in mean PAP via an NO-dependent mechanism. These data open up new avenues for the clinical management of PAH.

Pulmonary hypertension is associated with a significant reduction in pulmonary artery pulsatility, contractility and elasticity, contributing to an increase in pulmonary artery pressure and pulmonary resistance. Using a hypoxic piglet model, this study demonstrated that improving pulmonary artery plasticity using a newly developed pulsatile catheter improves hypoxic pulmonary hypertension.

增加肺动脉脉动流改善缺氧性肺动脉高压的仔猪

Pulmonary arterial hypertension (PAH) is a disease affecting distal pulmonary arteries (PA). These arteries are deformed, leading to right ventricular ...

Transthoracic Speckle Tracking Echocardiography for the Quantitative Assessment of Left Ventricular Myocardial Deformation

The value of conventional echocardiography is limited by differences in inter-individual image interpretation and therefore largely dependent on the examiners&#39; expertise. Speckle tracking Echocardiography (STE) is a promising but technically challenging method that can be used to quantitatively assess regional and global systolic and diastolic myocardial performance. Myocardial strain and strain rate can be measured in all three dimensions&#160;&#8212; radial, circumferential, longitudinal &#8212; of myocardial deformation. Standard cross-sectional two-dimensional B-mode images are recorded and subsequently postprocessed by automated continuous frame-by-frame tracking and motion analysis of speckles within the myocardium. Images are recorded as digital loops and synchronized to a 3-lead EKG for timing purposes. Longitudinal deformation is assessed in the apical 4-, 3-, and 2-chamber views. Circumferential and radial deformation are measured in the parasternal short axis plane.
Optimal image quality and accurate tissue tracking are paramount for the correct determination of myocardial performance parameters. Utilizing transthoracic STE in a healthy volunteer, the present article is a detailed outline of the essential steps and potential pitfalls of quantitative echocardiographic myocardial deformation analysis.

Speckle tracking echocardiography is an emerging diagnostic imaging technique for the quantitative assessment of global and regional myocardial performance. Standard view echocardiographic motion images are recorded and deformation parameters are subsequently measured by automated continuous frame-by-frame tracking and motion analysis of speckles within the B-mode images of the myocardium.

经胸斑点跟踪超声心动图左室心肌变形的定量评价

The value of conventional echocardiography is limited by differences in inter-individual image interpretation and therefore largely dependent on the ...

Murine Left Anterior Descending (LAD) Coronary Artery Ligation: An Improved and Simplified Model for Myocardial Infarction

Ischemic heart disease (IHD), or acute coronary syndrome (ACS), is one of the leading causes of death in the United States. IHD is characterized by reduced blood supply to the heart, resulting in the loss of oxygen to and the ensuing necrosis of the heart muscle. The MI model has gained popularity for its use as a short-term ischemia-reperfusion model and a long-term permanent ligation model. Below, we describe a reliable method for the permanent ligation of the LAD. With mouse genetic engineering technology becoming more advanced, and with an increasing availability of quality murine surgical instruments, the mouse has become a popular model for MI surgeries. Our surgical model incorporates the use of an easily reversible anesthetic for the rapid recovery of the mouse; a minimally invasive endotracheal intubation without involving a tracheotomy; and a thoracentesis through the original thoracotomy site without creating an additional incision in the chest, as is done in some other methods, to effectively remove excess blood and air from the chest cavity. This method is comparatively less invasive than other methods, which dramatically reduces surgical and post-surgical complications and mortality and improves reproducibility.

Murine Left Anterior Descending LAD Coronary Artery Ligation: An Improved and Simplified Model for Myocardial Infarction

We provide a reliable method for left anterior descending artery (LAD) ligation in a mouse model. This method is comparatively less invasive than other methods, involving endotracheal intubation, a left-sided thoracotomy approach, and thoracentesis. This method can be used as a model for both acute and chronic myocardial infarction (MI).

鼠左前降支（LAD）冠状动脉结扎:改进和简化模型的心肌梗死

Ischemic heart disease (IHD), or acute coronary syndrome (ACS), is one of the leading causes of death in the United States. IHD is characterized by ...

Direct Re-implantation of Left Coronary Artery into the Aorta in Adults with Anomalous Origin of Left Coronary Artery from the Pulmonary Artery (ALCAPA)

Anomalous origin of the left coronary artery from the pulmonary artery (ALCAPA) is a rare congenital anomaly which is one of leading causes of myocardial ischemia and infarction in children. If left untreated, it results in a 90% mortality rate in the first year of life. In patients who survive to the adulthood, the coronary steal phenomenon and retrograde left-sided coronary flow provide a substrate for chronic subendocardial ischemia, which may lead to left ventricular dysfunction, ischemic mitral regurgitation, malignant ventricular arrhythmias, and sudden cardiac death. The average age of life-threatening presentation is 33 years and of sudden cardiac death 31 years. Therefore, surgical correction is highly recommended as soon as the diagnosis is made, regardless of age. In adult-type ALCAPA originating from the right-facing sinus of the pulmonary artery, direct re-implantation of the ALCAPA into the aorta is the more physiologically sound repair technique to re-establish the dual-coronary perfusion system and is recommended. This protocol describes the technique of direct re-implantation of adult-type ALCAPA into the aorta.

Direct Re-implantation of Left Coronary Artery into the Aorta in Adults with Anomalous Origin of Left Coronary Artery from the Pulmonary Artery ALCAPA

Surgical correction of ALCAPA is highly recommended, regardless of age or the degree of intercoronary collateralization. This protocol presents a technique for the direct re-implantation of adult-type ALCAPA into the aorta to re-establish the dual-coronary perfusion. Whenever feasible, direct re-implantation is preferred to other surgical correction techniques.

左冠状动脉直接再注入与从肺动脉左冠状动脉起源异常成人主动脉（ALCAPA）

Anomalous origin of the left coronary artery from the pulmonary artery (ALCAPA) is a rare congenital anomaly which is one of leading causes of myocardial ...

Left Atrial Stenosis Induced Pulmonary Venous Arterialization and Group 2 Pulmonary Hypertension in Rat

The mechanism of mitral stenosis-induced pulmonary venous arterialization and group 2 pulmonary hypertension (PH) is unclear. There is no rodent model of group 2 PH, due to mitral stenosis (MS), to facilitate the investigation of disease mechanisms and potential therapeutic strategies. We present a novel rat model of pulmonary venous congestion-induced pulmonary venous arterialization and group 2 PH caused by left atrial stenosis (LAS). LAS is achieved by constricting the left atrium using a half-closed titanium clip. After the LAS surgery, a rat model with a transmitral inflow velocity greater than or equal to 2.0 m/s on echocardiography gradually develops pulmonary venous arterialization and group 2 PH over an 8- to 10-week period. In this protocol, we provide the step-by-step procedure of how to perform the LAS surgery. The presented LAS rat model mimics MS in humans and is useful for studying the underlying molecular mechanism of pulmonary venous arterialization and for the preclinical evaluation of therapies for group 2 PH.

Left atrial stenosis (LAS) is a novel surgical technique used for studying group 2 pulmonary hypertension (PH) and mechanisms underlying pulmonary venous arterialization. Here, we present a protocol to constrict the left atrium using a titanium clip to cause pulmonary venous arterialization and moderate PH in a rat.

左心房狭窄致大鼠肺静脉动脉化及第2组肺动脉高压

The mechanism of mitral stenosis-induced pulmonary venous arterialization and group 2 pulmonary hypertension (PH) is unclear. There is no rodent model of ...

Murine Myocardial Infarction Model using Permanent Ligation of Left Anterior Descending Coronary Artery

Myocardial infarction (MI) and acute coronary diseases are among the most prominent causes of death in population with western lifestyle. The murine models of MI with permanent ligation of left-anterior descending (LAD) coronary artery closely mimics MI in humans. Murine models benefit from the extensive genetic engineering available nowadays. Here we propose a reproducible murine surgical model of myocardial infarction by permanent LAD coronary ligation. Our technique comprises anesthesia with ketamine/xylazine that can be rapidly reversed by administration of an antagonist, intubation without tracheotomy for mechanical-assisted ventilation, ventilation with application of extrinsic positive end-expiratory pressure (PEEP) to avoid alveolar collapse, a thoracotomy method limiting to the minimum surgical lesions made to skeletal muscles, and lung inflation without thoracentesis. This method is sparsely invasive, reproducible and reduces post-surgery mortality and complications.

Herein we describe a surgical procedure showing how to achieve permanent ligation of the left-anterior descending coronary artery in mice. This model is of high relevance to investigate the pathophysiology of myocardial infarction and the concomitant biological processes.

使用左前部降冠动脉的永久结扎的月心肌梗死模型

Myocardial infarction (MI) and acute coronary diseases are among the most prominent causes of death in population with western lifestyle. The murine ...