Name: technique of minimally invasive transverse aortic constriction in mice for induction of left ventricular hypertrophy
Uploaded: 2017-09-25T12:30:00
Description: Watch this Scientific Journal Video about Technique of Minimally Invasive Transverse Aortic Constriction in Mice for Induction of Left Ventricular Hypertrophy at JoVE.com

这项工作得到了瑞士心血管基金会的赠款 (编号 32016) 的支持。

该协议中使用了 雄性 C57BL/6J 鼠 (10 周, 25-30g, n=60)。根据法国农业部和高等教育和研究部制定的准则, 动物接受人道护理, 所有程序都按照欧洲共同体理事会1986年11月24日的指示执行 (86/609/欧洲经济共同体) 和法国法律。该议定书得到了 #34 的批准; 动物实验 CREMEAS 和 #34 区域伦理委员会; (#2016092816207606). 
 1. 手术准备 <ol> <li> 在到达动物设施后一周内, 在标准的笼子里, 用12小时/1...

<ol>
	<li>Molinari, F., Malara, N., Mollace, V., Rosano, G., Ferraro, E. <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+cardiac+cachexia.">Animal models of cardiac cachexia.</a> Int. J. Cardiol. 219 (15), 105-110 (2016).</li><li>Tarnavski, O. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mouse+surgical+models+in+cardiovascular+research.">Mouse surgical models in cardiovascular research.</a> Methods. Mol. Biol. 573, 115-137 (2009).</li><li>Verma, S. K., Krishnamurthy, P., Kishore, R., Ardehali, 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=Transverse+aortic+constriction:+a+model+to+study+heart+failure+in+small+animals.">Transverse aortic constriction: a model to study heart failure in small animals.</a> Manual of Research Techniques in Cardiovascular Medicine. , 164-169 (2014).</li><li>Patten, R. D., Hall-Porter, M. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Small+animal+models+of+heart+failure.+Development+of+novel+therapies,+past+and+present.">Small animal models of heart failure. Development of novel therapies, past and present.</a> Circ. Heart Fail. 2 (2), 138-144 (2009).</li><li>Rockman, H. 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=Segregation+of+atrial-specific+and+inducible+expression+of+an+atrial+natriuretic+factor+transgene+in+an+in+vivo+murine+model+of+cardiac+hypertrophy.">Segregation of atrial-specific and inducible expression of an atrial natriuretic factor transgene in an in vivo murine model of cardiac hypertrophy.</a> Proc. Natl. Acad. Sci. USA. 88 (18), 8277-8281 (1991).</li><li>Barrick, C. J., Rojas, M., Schoonhoven, R., Smyth, S. S., Threadgill, D. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cardiac+response+to+pressure+overload+in+129S1/SvImJ+and+C57BL/6J+mice:+temporal-+and+background-dependent+development+of+concentric+left+ventricular+hypertrophy.">Cardiac response to pressure overload in 129S1/SvImJ and C57BL/6J mice: temporal- and background-dependent development of concentric left ventricular hypertrophy.</a> Am. J. Physiol. Heart Circ. Physiol. 292 (5), 2119-2130 (2007).</li><li>Deschepper, C. F., Olson, J. L., Otis, M., Gallo-Payet, N. <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+blood+pressure+and+morphological+traits+in+cardiovascular-related+organs+in+13+different+inbred+mouse+strains.">Characterization of blood pressure and morphological traits in cardiovascular-related organs in 13 different inbred mouse strains.</a> J. Appl. Physiol. 97 (1), 369-376 (2004).</li><li>Almeida, A. C., van Oort, R. J., 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=Transverse+aortic+constriction+in+mice.">Transverse aortic constriction in mice.</a> J. Vis. Exp. (38), (2010).</li><li>Hu, P., Zhang, D., Swenson, L., Chakrabarti, G., Abel, E. D., Litwin, S. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Minimally+invasive+aortic+banding+in+mice:+effects+of+altered+cardiomyocyte+insulin+signaling+during+pressure+overload.">Minimally invasive aortic banding in mice: effects of altered cardiomyocyte insulin signaling during pressure overload.</a> Am. J. Physiol. Heart Circ. Physiol. 285 (3), 1261-1269 (2003).</li><li>Faerber, G., 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+heart+failure+by+minimally+invasive+aortic+constriction+in+mice:+Reduced+peroxisome+proliferator-activated+receptor+&#978;+coactivator+levels+and+mitochondrial+dysfunction.">Induction of heart failure by minimally invasive aortic constriction in mice: Reduced peroxisome proliferator-activated receptor &#978; coactivator levels and mitochondrial dysfunction.</a> J. Thorac. Cardiovasc. Surg. 141 (2), 492-500 (2011).</li><li>Andersen, N. M., Tang, R., Li, L., Javan, H., Zhang, X. Q., Selzman, C. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=IKK-&#946;+inhibition+prevents+adaptive+left+ventricular+hypertrophy.">IKK-&#946; inhibition prevents adaptive left ventricular hypertrophy.</a> J. Surg. Res. 178 (1), 105-109 (2012).</li><li>Nemska, S., Monassier, L., Gassmann, M., Frossard, N., Tavakoli, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Kinetic+mRNA+profiling+in+a+rat+model+of+left+ventricular+hypertrophy+reveals+early+expression+of+chemokines+and+their+receptors.">Kinetic mRNA profiling in a rat model of left ventricular hypertrophy reveals early expression of chemokines and their receptors.</a> PLoS ONE. 11 (8), 0161273 (2016).</li><li>Liao, Y., 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=Echocardiographic+assessment+of+LV+hypertrophy+and+function+in+aortic-banded+mice:+necropsy+validation.">Echocardiographic assessment of LV hypertrophy and function in aortic-banded mice: necropsy validation.</a> Am. J. Physiol. Heart Circ. Physiol. 282 (5), 1703-1708 (2002).</li><li>Stansfield, W. E., 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+a+model+to+independently+study+regression+of+ventricular+hypertrophy.">Characterization of a model to independently study regression of ventricular hypertrophy.</a> J. Surg. Res. 142 (2), 387-393 (2007).</li><li>Beetz, 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=Ablation+of+biglycan+attenuates+cardiac+hypertrophy+and+fibrosis+after+left+ventricular+pressure+overload.">Ablation of biglycan attenuates cardiac hypertrophy and fibrosis after left ventricular pressure overload.</a> J. Mol. Cell. Cardiol. 101, 145-155 (2016).</li></ol>

本议定书的目的是提出一个 step-by 的外科技术, 微侵袭横动脉狭窄小鼠。对小鼠横主动脉狭窄的详细技术描述已由其他作者2,8报告。然而, 这些研究者在动物插管和通气后进行手术。使用额外的一步插管通气增加了整个过程的复杂性和持续时间和全球压力的动物接触。因此, 微创横脉狭窄的概念得到了一定的重视。小鼠的微创横动脉狭窄用于诱发左心室?...

在过去的几年中, 心力衰竭的研究已经在可行的动物模型中进行了1。与大型动物模型的心力衰竭相比, 小动物模型有许多潜在的优势。除了低成本的住房和维修, 小动物模型可以访问更多的研究人员, 由于不太复杂的设施需要2。
小鼠心力衰竭模型提供了许多相同的优势, 与老鼠模型。除了减少的住房成本3, 鼠标模型受益于相关的转基因和淘汰赛 (KO) 菌株的可用性。细胞类型特异、诱导性 KO 或转基因策略的可能性使小鼠成为研究心力衰竭发病机制的宝贵工具, 并尝试识别新的治疗方案3。
在目前使用的心力衰竭的小鼠模型中4中, 首次用人5描述的横主动脉收缩 (TAC) 是产生压力超负荷引起的左心室肥厚 (肥厚)1的首选模型。,3. 该模型的最大优点是能够允许左心室肥厚的分层, 尽管在不同的小鼠品系中, 对 TAC 的反应是可变的.特别是, C57BL/6 小鼠发展快速 LV 扩张后, TAC 可能不会发生其他菌株4,6,7。
在2周内, 以 TAC 引起的高血压的突然发作使 LV 肿块增加约 50%, 从而可以快速检查旨在调节左心室肥厚4发育的药理或分子干预活动。急性高血压的 TAC 不完全再现左心室肥厚和重构观察在主动脉狭窄或动脉高血压的临床设置。然而, 该模型被许多研究者用来识别和修改心力衰竭的新治疗指标4。
执行 TAC 在小鼠需要更大的手术经验比其他技术所需的诱导肥厚和随后心力衰竭2。大多数作者执行此过程的插管和通风的动物2,8, 这使得这个过程更苛刻和耗时, 并增加了对动物的外科负担。只有很少的调查人员在他们的研究中使用了微创的 TAC, 简要地参考了手术过程9,10,11。
本议定书的目的是描述 step-by 一步简化和 user-friendly 的小鼠横主动脉狭窄的技术, 突出了程序的关键阶段。通过遵循这些关键步骤, 可以轻松地执行此项技术。

<table width="556">
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	<tbody>
		<tr height="20">
			<td height="20" style="width:216px;height:20px;">Surgical microscope</td>
			<td style="width:108px;">Olympus</td>
			<td style="width:171px;">SZX2-TR30</td>
			<td style="width:63px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Razor</td>
			<td>Rowenta</td>
			<td>Nomad TN3650FO</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="width:495px;height:20px;">Sutures:</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Polypropylene 7/0</td>
			<td>Ethicon</td>
			<td>BV-1X</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Polypropylene 6/0</td>
			<td>BBraun</td>
			<td>C0862061</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Silk 6/0 ligature&#160;</td>
			<td>FST</td>
			<td>18020-60</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Polypropylene 4/0</td>
			<td>Ethicon</td>
			<td>8683</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Polypropylene 5/0</td>
			<td>Ethicon</td>
			<td>Z303</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="width:495px;height:20px;">Drugs:</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Ketamin</td>
			<td>Merial</td>
			<td>Imalg&#232;ne 1000, LBM154AD</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Xylazine</td>
			<td>Bayer</td>
			<td>Rompun 2%, KP09PPC</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Buprenorphine</td>
			<td>Ceva</td>
			<td>Vetergesic, 072013</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="width:495px;height:20px;">Instruments:&#160;</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Bone nippers</td>
			<td>Fine Surgical Tools</td>
			<td>16101-10</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Ligation aid</td>
			<td>Fine Surgical Tools</td>
			<td>18062-12</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Tying forceps</td>
			<td>Fine Surgical Tools</td>
			<td>18026-10</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Needle holder Crile-Wood</td>
			<td>Fine Surgical Tools</td>
			<td>12003-15</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Microsurgery forceps&#160;</td>
			<td>Fine Surgical Tools</td>
			<td>11003-12</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Microsurgery forceps&#160;</td>
			<td>Fine Surgical Tools</td>
			<td>11002-12</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Tissue forceps</td>
			<td>Fine Surgical Tools</td>
			<td>11021-12</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Microsurgery needle holder</td>
			<td>Fine Surgical Tools</td>
			<td>12076-12</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Microsurgery scissors</td>
			<td>Fine Surgical Tools</td>
			<td>91501-09</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Mayo scissors</td>
			<td>Fine Surgical Tools</td>
			<td>14511-15</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">11-blade knife</td>
			<td>Fine Surgical Tools</td>
			<td>10011-00</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="width:495px;height:20px;">RNA extraction and qPCR:</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">TriReagent</td>
			<td>Euromedex</td>
			<td>TR-118-200</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Rneasy Mini kit</td>
			<td>Qiagen</td>
			<td>74704</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Qubit Fluorimetric RNA assay</td>
			<td>Fisher Scientific</td>
			<td>10034622</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">RNA 6000 Nano kit</td>
			<td>Agilent</td>
			<td>5067-1511</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">High Capacity cDNA kit</td>
			<td>Fisher Scientific</td>
			<td>10400745</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Taqman Master Mix</td>
			<td>Fisher Scientific</td>
			<td>10157154</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Taqman BNP primers</td>
			<td>Fisher Scientific</td>
			<td>Mm01255770_g1</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="width:216px;height:20px;">Taqman ANP primers</td>
			<td>Fisher Scientific</td>
			<td>Mm01255747_g1&#160;</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Taqman ACE primers</td>
			<td>Fisher Scientific</td>
			<td>Mm00802048_m1</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Taqman Col1a1 primers&#160;</td>
			<td>Fisher Scientific</td>
			<td>Mm00801666_g1</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Taqman TGFb primers</td>
			<td>Fisher Scientific</td>
			<td>Mm01178820_m1</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Taqman Gapdh primers</td>
			<td>Fisher Scientific</td>
			<td>Mm99999915_g1</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">ABIPrism&#160; Thermocycler</td>
			<td>Applied Biosystems</td>
			<td>7000</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="width:495px;height:20px;">Software:</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">GraphPad Prism</td>
			<td>GraphPad</td>
			<td>Prism 7</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="width:495px;height:20px;">Animal food</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr height="27">
			<td height="27" style="width:216px;height:27px;">Complete diet for adult rats/mice</td>
			<td style="width:108px;">Safe</td>
			<td style="width:171px;">UB220610R</td>
			<td></td>
		</tr>
	</tbody>
</table>

technique of minimally invasive transverse aortic constriction in mice for induction of left ventricular hypertrophy

小鼠横主动脉狭窄 (TAC) 是压力超负荷所致左心室肥厚的实验研究和心力衰竭进展的最常用的外科技术之一。在大多数报告的调查中, 这一程序是用插管和通气的动物进行的, 这使得它要求和耗时, 并增加了对动物的外科负担。本协议的目的是描述一个简单的技术, 无插管和通气小鼠微创 TAC。强调了该技术的关键步骤, 以达到低死亡率和高效率的诱导肥厚。
雄性 C57BL/6 小鼠 (10 周大, 25-30 克, n=60) 被麻醉, 单腹腔注射氯胺酮和嗪的混合物。在一个自发呼吸的动物在3-4 毫米上部部份胸骨之后, 6/0 丝缝线穿线通过结扎援助的眼睛通过在主动脉弓之下和栓在一条钝的27测量针。假手术的动物进行了相同的外科准备, 但没有主动脉收缩。该程序在诱导肥厚中的疗效证实了心脏/体重比的显著增加。这个比率在 3, 7, 14 和28在手术以后获得 (n = 6-10 在每个小组和每个时间点)。运用我们的技术, 在 TAC 中观察到7天28的假动物的肥厚。手术和晚 (超过28天) 死亡率都非常低, 在1.7%。
总之, 我们的小鼠微创 cost-effective 技术具有非常低的手术和术后死亡率, 是高效的诱导肥厚。它简化了手术过程, 减少了对动物的压力。通过遵循本协议中描述的关键步骤, 可以很容易地执行该操作。

手术和晚期生存 手术的生存是非常高的, 98.3% (59 出 60) 为整个系列 (TAC 和假操作的动物)。唯一的手术死亡是由于一个小鼠的出血并发症, 计划为假手术。术后存活28天的观察期也很好, 98.3% (58 出 59)。唯一的晚术后死亡发生在一天 (D) 16, 可能是心脏起源的 TAC 鼠标。
技术验证 所提出的技术是非常可?...

Watch this Scientific Journal Video about Technique of Minimally Invasive Transverse Aortic Constriction in Mice for Induction of Left Ventricular Hypertrophy at JoVE.com

Technique of Minimally Invasive Transverse Aortic Constriction in Mice for Induction of Left Ventricular Hypertrophy

本议定书的目的是描述 step-by 微创的横向主动脉狭窄 (TAC) 技术的小鼠。通过消除气管插管和通气是常用的标准程序的强制性, 微创 TAC 简化了手术程序, 减少了对动物的压力。

小鼠横主动脉狭窄诱导左心室肥厚的技术研究

Transverse aortic constriction (TAC) in mice is one of the most commonly used surgical techniques for experimental investigation of pressure ...

The overall goal of this protocol is to establish minimally invasive transverse aortic constriction in mice. This method can help answer key questions about implementing transverse aortic constriction in mice, such as how to create a passage under the aortic arch. The main advantages of this technique are that it is simple, user-friendly, and that it eliminates the need for intubating and ventilating the mice.I will be demonstrating the procedure with our postdoc Simona Nemska who will be performing the presurgical animal preparation and postoperative surveillance steps. On the day of the procedure, shave the neck and chest of an anesthetized mouse and disinfect the exposed skin with 70%ethanol. After confirming the appropriate level of sedation by toe pinch, secure the animal in the supine position onto a clean cork working pad.Next, use a number 11 knife blade to make a 10 millimeter longitudinal midline cervical incision from the suprasternal notch to the mid chest to expose the sternum. Using a Crile-Wood needle holder, pass a 4-0 monofilament polypropylene stay suture to retract the thyroid. And tape the suture to the working pad.Then move the working pad under a surgical microscope and use microsurgical forceps to bluntly separate the pretracheal muscles to uncover the trachea. Gently slide the closed tips of a pair of curved microsurgical forceps over the trachea and behind the sternum and carefully open and close the curved forceps to bluntly dissect the pleura. Use the straight microsurgical forceps to grasp the right supraclavicular muscles and gently pull up on the chest.Sliding the inferior tip of the bone nipper under the sternum, perform a three to four millimeter upper partial sternotomy, directing the lower part of the mini sternotomy slightly toward the left. Using a microsurgical needle holder, pass a 7-0 monofilament polypropylene stay suture from the inside to the outside of the second intercostal space on each side of the mini sternotomy. Use 7-0 monofilament polypropylene stay sutures to spread the sternal edges on each side.And use the curved forceps to gently move the pretracheal muscles, mediastinal fat, and thymus aside to visualize the aortic arch under low power magnification. To expose the soft tissue under the aortic arch, insert tying forceps under the arch with the tips closed and gently open the forceps. Use a second tying forceps to make a tunnel in the soft tissue under the aortic arch by gently opening and closing the forceps in the soft tissue.Then pass a piece of 6-0 silk ligature threaded through the eye of a ligation aid under the aortic arch, using a tying forceps to retrieve the suture from between the origin of the right innominate and left common carotid arteries. The singular most difficult step of the procedure is inserting the suture under the aortic arch. Taking adequate care while creating the tunnel under the arch will make the subsequent suture placement easier.Now use the straight forceps to position a blunted 27 gauge needle next to the aortic arch and use the two tying forceps to tie the suture snugly around the needle and the aorta between the right innominate and left common carotid arteries. When the suture has been secured, quickly but gently remove the needle to achieve a 0.4 millimeter diameter narrowing and a reproducible transverse 65 to 70%aortic constriction. After confirming hemostasis, remove the 7-0 monofilament polypropylene stay sutures used for spreading the sternal edges.Use a microsurgical needle holder to pass a simple 6-0 monofilament polypropylene suture from the outside to the inside of the left second intercostal space and from the inside to the outside of the right second intercostal space. Use the Crile-Wood needle holder to bring the sternal edges together with a 6-0 monofilament polypropylene suture. And close the skin with a 5-0 monofilament polypropylene running suture in one layer with a Crile-Wood needle holder.Then place the animal back in its cage in the prone position under warming with monitoring until full recovery. The efficacy of the left ventricular hypertrophy induction can be validated by comparing the heart weight to body weight ratios of the sham animals to that of the experimental animals at days three, seven, 14, and 28 postsurgery. The ratio significantly increases in the banded animals compared to the sham groups from postoperative day seven on, remaining significantly higher in the constricted animals up through day 28 postsurgery.The observed increase in the heart to body weight ratios is due solely to a rise in the left ventricle body weight ratio as the right ventricle body weight ratio remains comparable between the transverse aortic constriction and the sham operated animals over the entire observation period. In addition, the mRNA expression of the brain natriuretic peptide, a positive control for cardiac hypertrophy in the left ventricles, is significantly higher in the aortic banded animals compared to the control sham operated animals at day 14, further validating the efficiency of the technique. Once mastered, this technique can be completed in 20 minutes if it is performed properly.While attempting this procedure, it is important to use a ligation aid and tying forceps similar to those used in our protocol. After its development, this technique provided a reliable and simplified model for exploring the mechanisms involved in the genesis of pressure overload induced left ventricular hypertrophy. After watching this video, you should have a good understanding of how to perform minimally invasive transverse aortic constriction in mice with a special emphasis on the passage under the aortic arch.Don't forget that creating the passage under the aortic arch is the most critical step of the procedure, and using a ligation aid and tying forceps considerably reduces the risk of injury to the arch.

Research

JoVE Journal

Medicine

Ascending Aortic Constriction in Rats for Creation of Pressure Overload Cardiac Hypertrophy Model

大鼠建立压力超负荷心肌肥厚模型在升主动脉缩窄

Ascending aortic constriction is the most common and successful surgical model for creating pressure overload induced cardiac hypertrophy and heart ...

科研

医学

Assessment of Cardiac Morphological and Functional Changes in Mouse Model of Transverse Aortic Constriction by Echocardiographic Imaging

在横向主动脉缩窄的小鼠模型和心脏形态学评估功能改变通过超声心动图成像

Transverse aortic constriction (TAC) in mice has been used as a valuable model to study mechanisms of cardiac hypertrophy and heart failure1. A reliable ...

Minimally Invasive Transverse Aortic Constriction in Mice

微创横向主动脉缩窄小鼠

Minimally invasive transverse aortic constriction (MTAC) is a more desirable method for the constriction of the transverse aorta in mice than standard ...

A Minimally Invasive Model to Analyze Endochondral Fracture Healing in Mice Under Standardized Biomechanical Conditions

标准化生物力学条件下小鼠软骨骨折愈合的微创模型分析

Bone healing models are necessary to analyze the complex mechanisms of fracture healing to improve clinical fracture treatment. During the last decade, an ...

A Closed-chest Model to Induce Transverse Aortic Constriction in Mice

诱导小鼠横向主动脉狭窄的闭合胸模型

Research on cardiac hypertrophy and heart failure is frequently based on pressure overload mouse models induced by TAC. The standard procedure is to ...

Studying Left Ventricular Reverse Remodeling by Aortic Debanding in Rodents

研究左心室反向改造由主动脉脱体在啮齿动物

To better understand the left ventricular (LV) reverse remodeling (RR), we describe a rodent model wherein, after aortic banding-induced LV remodeling, ...

Invasive Hemodynamic Assessment for the Right Ventricular System and Hypoxia-Induced Pulmonary Arterial Hypertension in Mice

小鼠右心室系统及缺氧诱发肺动脉高血压的侵入性血液动力学评估

Pulmonary arterial hypertension (PAH) is a chronic and severe cardiopulmonary disorder. Mice are a popular animal model used to mimic this disease. ...

A Pre-clinical Rat Model for the Study of Ischemia-reperfusion Injury in Reconstructive Microsurgery

重建显微外科缺血再灌注损伤研究的临床前大鼠模型

Ischemia-reperfusion injury is the main cause of flap failure in reconstructive microsurgery. The rat is the preferred preclinical animal model in many ...

Operating Transverse Aortic Constriction with Absorbable Suture to Obtain Transient Myocardial Hypertrophy

操作横向主动脉收缩与可吸收的缝合获得瞬时心肌萎缩

Based on twice transverse aortic constrictions (TACs) in mice, it is proved that myocardial hypertrophic preconditioning (MHP) could attenuate ...

Echocardiographic Characterization of Left Ventricular Structure, Function, and Coronary Flow in Neonate Mice

新生小鼠左心室结构、功能和冠状动脉血流的超声心动图表征

Echocardiography is a non-invasive procedure that enables the evaluation of structural and functional parameters in animal models of cardiovascular ...