The authors would like to acknowledge the Cincinnati Children&#8217;s Hospital Division of Veterinary Services for their assistance. This manuscript was supported by the National Institutes of Health K12 HD028827. Anna Perl PhD taught the authors this surgical procedure.

注:动物使用声明:在本研究中的所有程序均经批准，并按照机构动物使用及护理委员会（IACUC）在辛辛那提儿童医院的指导方针进行。八周龄C57BL / 6J雄性小鼠从Jackson实验室（巴港，ME）中获得，并使其适应环境，使用前一个星期。直到手术，将动物饲养在无病原体的阻挡设施，并提供高压灭菌食物和过滤水的广告libdium。每个鼠笼用专用的空气和水的供给，并提供维持在12小时昼夜周期。以下从手术中恢复，小鼠保持在笼子里与过滤上衣，提供蒸压州城广告libidum，并提供了从一瓶水过滤水。 1.准备仪器<ol><li>使用曲别针和大头针6皮肤拉钩。扭曲拉直回形针在纸张的针柄，升屋檐5cm的直钢丝的一端，使单个0.5厘米"U"形钩在导线的末端。 </li><li>使用保鲜膜约15×15厘米见方的手术单。准备每只小鼠1敷料。把一纸塔在每个包装之间。 </li><li>消毒所有的手术工具以及一叠12×12英寸的软木砖，纱布，棉花拭子。 </li></ol> 2.鼠标准备<ol><li>诱导麻醉，用2％异氟醚。称量动物。 </li><li>在一个专门的手术准备区刮胡子左胸部和颈部电气剃须刀。 </li><li>申请一滴人工泪液软膏鼠标的眼睛。 </li><li>净化颈部和左胸部与洗必泰和异丙醇。重复两次以上。 </li></ol> 3.鼠标奥罗气管插管和机械通气<ol><li>有一个非无菌手术技术人员代替鼠标仰卧在预热手术区。</li><li>通过记录缺乏爪子掐的响应确认麻醉深度。 </li><li>洗手，并穿上手术装束，口罩，帽子后，穿上无菌手术手套。 </li><li>立体裁剪，并使用无菌技术后，使超过前中期的脖子，露出喉部有1厘米的垂直切口。轻轻缩回带状肌与弯曲的，锯齿10厘米钳，并通过与一个直剪刀的前端扩展的带状肌暴露喉和气管。 </li><li>口服插入一个22克钝尖血管导管插入中期气管（ 图1A）和目视确认放置（ 图1B）。维持麻醉和通风用1-3％异氟醚通过啮齿动物呼吸机（225微升每冲程;每分钟200斯托克斯）。采用15厘米H 2 O的压力极限</li></ol> 4.鼠标肺切除<ol><li>莱鼠标在右侧卧位，用鼠标的背面朝上操作员（左侧上）。用自密封的保鲜膜作为无菌的悬垂性。切割通过悬垂性好，用钝尖剪子弯，使长切割平行于肋骨2厘米，在第 4和第 5肋间。将钝头剪刀弯曲和底层的肋骨和肋间肌肉解剖皮肤了。 </li><li>缩回四个拉钩皮肤暴露正方形1.5×1.5cm的外科窗口（ 图2A）。固定拉钩，以软木板上。 </li><li>解剖下来使用弯钳肋，并使用弯钳之一尖端进入胸腔。 </li><li>使用钝头微型剪刀，使用下刀片进入胸腔。使肋和重复在相反的方向之间为0.5厘米的切口。 </li><li>使用剩下的两个拉钩，在前后轴和安全的拉钩到软木板上（ 图2B）打开胸腔。 </li> &lt;李&gt;使用在左手弯曲钝尖镊子，把握左肺和置换左肺的上部横向和下方通过开胸直到左肺动脉与支气管露出（ 图3A，B）。 </li><li>持在右手装入钛血管microclip敷贴与涂抹在手掌和弯曲尖端从手掌（ 图3C）背向的主体中，沿左侧的后方面的曲率滑动施用尖端进入胸腔肺和剪辑左支气管和肺动脉（ 图3D）。 </li><li>取下敷贴但保留左肺缩回。抓住钝尖微型剪刀用右手切支气管和肺动脉远端剪辑，删除左肺（ 图3E）。 </li><li>拆下肋骨拉钩。 </li><li>使用弯曲钝钳捏起来1厘米皮肤逊色于incisi上，但上述隔膜的水平，并插入穿过皮肤并进入左胸腔（ 图4A，B）的一个24克血管导管。 </li><li>使用5-0 prolene线缝合周围放置第 4 次 2间断缝合和第 5肋关闭胸腔。 </li><li>去除皮肤拉钩。使用两套镊子来近似沿切口的长度皮肤和粘在皮肤闭合。 </li><li>一个3毫升路厄锁注射器连接到血管导管和通过施加轻柔抽吸抽出血管导管除去残留的空气。 </li><li>胶颈部切口闭合使用镊子两套像以前一样。 </li></ol> 5.鼠标深水肺切除<ol><li>暴露左肺如"老鼠肺切除"协议指出。提起肋笼具有弯曲钝钳，以允许空气进入左胸腔（ 图5A，B）。 </li><li>放置一个24克血管导管插入左寿拉契奇腔上面，小心不要弄伤左肺。 </li><li>使用5-0普理灵缝合小心不要刺破肺（ 图5C），将两个长度的缝合材料到3 次 / 4和第 5/6 次肋骨间隙（ 图5D）。绑减轻左肺症的风险之前，将缝合材料的两个长度。领带的缝合材料，以使2中断线圈（ 图5E）。 </li><li>胶上的皮肤胸切开，除去残留的空气与血管导管，和胶水颈部切口如上。 </li></ol> 6.复苏，镇痛和恢复<ol><li>关掉异氟烷，并管理0.1毫克/公斤丁丙诺啡和0.5生理盐水皮下注射毫升。 </li><li>当自主呼吸恢复，取出气管插管。 </li><li>观察鼠标，直到它再次走动。通常走resum上课除去气管导管后几分钟。 </li><li>放置在27℃培养箱（加湿，25％氧气）鼠标恢复O / N。 注意:我们放置食物与水润湿的上箱地板为手术后的第一个24小时的几个粒料。 </li><li>手术后每日三天两次施用0.1毫克/千克丁丙诺啡腹腔注射。小心不要处理的动物时，打开手术部位。 </li></ol> 7.鼠标监测<ol><li>手术后称量小鼠在1，3,5和7天。 </li></ol>

<ol>
	<li>Strueby, L., Thebaud, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Advances+in+bronchopulmonary+dysplasia.">Advances in bronchopulmonary dysplasia.</a> Expert review of respiratory medicine. , (2014).</li><li>Donn, S. M., Sinha, 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=Recent+advances+in+the+understanding+and+management+of+bronchopulmonary+dysplasia.">Recent advances in the understanding and management of bronchopulmonary dysplasia.</a> Seminars in fetal &amp; neonatal medicine. 14, 332 (2009).</li><li>Molen, T., Miravitlles, M., Kocks, J. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=COPD+management:+role+of+symptom+assessment+in+routine+clinical+practice.">COPD management: role of symptom assessment in routine clinical practice.</a> International journal of chronic obstructive pulmonary disease. 8, 461-471 (2013).</li><li>Seifert, A. 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=Skin+shedding+and+tissue+regeneration+in+African+spiny+mice+(Acomys).">Skin shedding and tissue regeneration in African spiny mice (Acomys).</a> Nature. 489, 561-565 (2012).</li><li>Vidal, P., Dickson, M. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Regeneration+of+the+distal+phalanx.+A+case+report.">Regeneration of the distal phalanx. A case report.</a> Journal of hand surgery. 18, 230-233 (1993).</li><li>Potter, P. C., Levine, M. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Bone+Regeneration+Following+Chronic+Suppurative+Osteitis+of+the+Distal+Phalanx.">Bone Regeneration Following Chronic Suppurative Osteitis of the Distal Phalanx.</a> Annals of surgery. 80, 728-729 (1924).</li><li>McKim, L. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Regeneration+of+the+Distal+Phalanx.">Regeneration of the Distal Phalanx.</a> Canadian Medical Association journal. 26, 549-550 (1932).</li><li>Brown, L. M., Rannels, S. R., Rannels, D. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Implications+of+post-pneumonectomy+compensatory+lung+growth+in+pulmonary+physiology+and+disease.">Implications of post-pneumonectomy compensatory lung growth in pulmonary physiology and disease.</a> Respir Res. 2, 340-347 (2001).</li><li>Holder, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Regeneration+and+compensatory+growth.">Regeneration and compensatory growth.</a> British medical bulletin. 37, 227-232 (1981).</li><li>Hsia, C. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Lessons+from+a+canine+model+of+compensatory+lung+growth.">Lessons from a canine model of compensatory lung growth.</a> Curr Top Dev Biol. 64, 17-32 (2004).</li><li>Butler, J. P., 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=Evidence+for+adult+lung+growth+in+humans.">Evidence for adult lung growth in humans.</a> N Engl J Med. 367, 244-247 (2012).</li><li>Konerding, M. 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=Spatial+dependence+of+alveolar+angiogenesis+in+post-pneumonectomy+lung+growth.">Spatial dependence of alveolar angiogenesis in post-pneumonectomy lung growth.</a> Angiogenesis. 15, 23-32 (2012).</li><li>Kho, A. T., Liu, K., Visner, G., Martin, T., Boudreault, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Identification+of+dedifferentiation+and+redevelopment+phases+during+postpneumonectomy+lung+growth.">Identification of dedifferentiation and redevelopment phases during postpneumonectomy lung growth.</a> Am J Physiol Lung Cell Mol Physiol. 305, 542-554 (2013).</li><li>Wang, W., Nguyen, N. M., Guo, J., Longitudinal Woods, J. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Noninvasive+Monitoring+of+Compensatory+Lung+Growth+in+Mice+after+Pneumonectomy+via+(3)He+and+(1)H+Magnetic+Resonance+Imaging.">Noninvasive Monitoring of Compensatory Lung Growth in Mice after Pneumonectomy via (3)He and (1)H Magnetic Resonance Imaging.</a> Am J Respir Cell Mol Biol. 49, 697-703 (2013).</li><li>Gibney, B. 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=Detection+of+murine+post-pneumonectomy+lung+regeneration+by+18FDG+PET+imaging.">Detection of murine post-pneumonectomy lung regeneration by 18FDG PET imaging.</a> EJNMMI research. 2, 48 (2012).</li><li>Rawlins, E. L., Perl, A. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+a'MAZE'ing+world+of+lung-specific+transgenic+mice.">The a'MAZE'ing world of lung-specific transgenic mice.</a> Am J Respir Cell Mol Biol. 46, 269-282 (2012).</li><li>Ochs, M., Muhlfeld, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Quantitative+microscopy+of+the+lung:+a+problem-based+approach.+Part+1:+basic+principles+of+lung+stereology.">Quantitative microscopy of the lung: a problem-based approach. Part 1: basic principles of lung stereology.</a> Am J Physiol Lung Cell Mol Physiol. 305, L15-22 (2013).</li><li>Ysasi, A. 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=Effect+of+unilateral+diaphragmatic+paralysis+on+postpneumonectomy+lung+growth.">Effect of unilateral diaphragmatic paralysis on postpneumonectomy lung growth.</a> Am J Physiol Lung Cell Mol Physiol. 305, L439-445 (2013).</li><li>Dane, D. M., Yilmaz, C., Estrera, A. S., Hsia, C. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Separating+in+vivo+mechanical+stimuli+for+postpneumonectomy+compensation:+physiological+assessment.">Separating in vivo mechanical stimuli for postpneumonectomy compensation: physiological assessment.</a> Journal of applied physiology. 114, 99-106 (2013).</li><li>Mortola, J. P., Magnante, D., Saetta, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Expiratory+pattern+of+newborn+mammals.">Expiratory pattern of newborn mammals.</a> Journal of applied physiology. 58, 528-533 (1985).</li><li>MacDonald, K. D., Chang, H. Y., Mitzner, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=An+improved+simple+method+of+mouse+lung+intubation.">An improved simple method of mouse lung intubation.</a> Journal of applied physiology. 106, 984-987 (2009).</li></ol>

The authors have nothing to disclose.

我们已经提供了迄今报告的鼠标PNX和鼠标假PNX程序的最详细的描述。我们已经取得的读者了解一些常见的陷阱，调查人员学习的过程通常会遇到，我们已经介绍了我们的实验室开发，以减轻对这些缺陷的几个技巧。利用这种模式其他实验室可能已经开发了其他的技术修改或使用不同的仪器。在评估的技术差异，个别研究者将需要决定是否这些差异显著影响实验结果。 补偿性?...

肺的主要功能是提供一个有机体和大气之间的氧气和二氧化碳交换。在人类中，一台主机的先天和后天条件导致降低的肺表面面积导致肺功能受损。虽然宿主疗法，如吸入皮质类固醇，支气管扩张剂，补充氧气和慢性机械通气用于减轻肺功能受损1-3的后果，提供了理想的治疗，这些条件将促进功能的肺组织的再生- 即，肺再生。 哺乳动物组织再生已经有据可查。非洲刺鼠可再生大面积皮肤无疤痕形成4。在人类的末节指骨可再生损伤后或截肢5-7。继肺（PNX），肺代偿出现增长小鼠8只9，做GS 10和人类11。根据定义，补偿性肺生长包括现有的空域与扩展相关的微循环12不仅扩大这些扩大空域，但重新septation。基因表达分析表明，这种模式概括很多肺发育13的信号事件。四周后小鼠PNX，肺泡表面面积相当于假手术的动物14。在这个手稿中，我们描述鼠标PNX和假PNX程序。

<table border="0" cellpadding="0" cellspacing="0" width="1052">
	<tbody>
		<tr height="22">
			<td height="22" style="height:22px;width:263px;">Name of the reagent</td>
			<td style="width:263px;">Company</td>
			<td style="width:263px;">Catalogue number</td>
			<td style="width:263px;">Comments (optional)</td>
		</tr>
		<tr height="101">
			<td height="101" style="height:101px;width:263px;">6-inch Vascular Clip Applicator</td>
			<td style="width:263px;">Teleflex Medical (WECK)</td>
			<td style="width:263px;">137062</td>
			<td style="width:263px;"></td>
		</tr>
		<tr height="101">
			<td height="101" style="height:101px;width:263px;">Horizon Small Titanium Red Clip</td>
			<td style="width:263px;">Teleflex Medical (WECK)</td>
			<td style="width:263px;">1201</td>
			<td style="width:263px;"></td>
		</tr>
		<tr height="121">
			<td height="121" style="height:121px;width:263px;">Narrow Pattern 12cm Curved Forceps</td>
			<td style="width:263px;">Fine Science Tools</td>
			<td style="width:263px;">11003-12</td>
			<td style="width:263px;"></td>
		</tr>
		<tr height="141">
			<td height="141" style="height:141px;width:263px;">Curved Serated 10 cm Graefe Forceps&#160;</td>
			<td style="width:263px;">Fine Science Tools</td>
			<td style="width:263px;">11052-10</td>
			<td style="width:263px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:263px;">Castroviejo Needle Holder</td>
			<td style="width:263px;">Fine Science Tools</td>
			<td style="width:263px;">12565-14</td>
			<td style="width:263px;"></td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;width:263px;">Straight 9 cm Strabismus Scissors (Blunt Tip)</td>
			<td style="width:263px;">Fine Science Tools</td>
			<td style="width:263px;">14075-09</td>
			<td style="width:263px;"></td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;width:263px;">Straight 8.5 cm Hardened Fine Scissors</td>
			<td style="width:263px;">Fine Science Tools</td>
			<td style="width:263px;">14090-09</td>
			<td style="width:263px;"></td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;width:263px;">Straight, Blunt Tip Cohan-Vannas Spring Scissors</td>
			<td style="width:263px;">Fine Science Tools</td>
			<td style="width:263px;">15000-12</td>
			<td style="width:263px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:263px;">Skin Glue</td>
			<td style="width:263px;">Gluture</td>
			<td style="width:263px;">32046</td>
			<td style="width:263px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:263px;">22 ga angiocatheter</td>
			<td style="width:263px;"></td>
			<td style="width:263px;"></td>
			<td style="width:263px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:263px;">24 ga angiocatheter</td>
			<td style="width:263px;"></td>
			<td style="width:263px;"></td>
			<td style="width:263px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:263px;">3 mL luer lock syringe</td>
			<td style="width:263px;"></td>
			<td style="width:263px;"></td>
			<td style="width:263px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:263px;">4 short retractors</td>
			<td style="width:263px;"></td>
			<td style="width:263px;"></td>
			<td style="width:263px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:263px;">2 long retractors</td>
			<td style="width:263px;"></td>
			<td style="width:263px;"></td>
			<td style="width:263px;"></td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;width:263px;">5-0 Prolene on curved cutting needle</td>
			<td style="width:263px;">Ethicon</td>
			<td style="width:263px;">8698G</td>
			<td style="width:263px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:263px;">0.5 mL syringe on 27 ga needle</td>
			<td style="width:263px;"></td>
			<td style="width:263px;"></td>
			<td style="width:263px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:263px;">Normal Saline</td>
			<td style="width:263px;"></td>
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mouse pneumonectomy model of compensatory lung growth

In humans, disrupted repair and remodeling of injured lung contributes to a host of acute and chronic lung disorders which may ultimately lead to disability or death. Injury-based animal models of lung repair and regeneration are limited by injury-specific responses making it difficult to differentiate changes related to the injury response and injury resolution from changes related to lung repair and lung regeneration. However, use of animal models to identify these repair and regeneration signaling pathways is critical to the development of new therapies aimed at improving pulmonary function following lung injury. The mouse pneumonectomy model utilizes compensatory lung growth to isolate those repair and regeneration signals in order to more clearly define mechanisms of alveolar re-septation. Here, we describe our technique for performing mouse pneumonectomy and sham pneumonectomy. This technique may be utilized in conjunction with lineage tracing or other transgenic mouse models to define molecular and cellular mechanism of lung repair and regeneration.

。PNX，假手术鼠标重量的情节在图6提供了在我们的手中，生存始终是95 - 100％为PNX和假肺。有关如何右肺再生长在该模型和预期的时间过程中，我们建议读者参考吉布尼等人的 15和Wang 等人的手稿的描述。14 几种常见的陷阱，必须避免成功执行鼠标PNX和鼠标假肺的程序。 气管插管 :许多研究者学习?...

Watch this Scientific Journal Video about Mouse Pneumonectomy Model of Compensatory Lung Growth at JoVE.com

Mouse Pneumonectomy Model of Compensatory Lung Growth

Mouse pneumonectomy is a commonly employed model of compensatory lung growth. This procedure can be used in conjunction with lineage tracing or transgenic mouse models to elucidate underlying mechanisms.

补偿性增长龙鼠标肺切除型号

In humans, disrupted repair and remodeling of injured lung contributes to a host of acute and chronic lung disorders which may ultimately lead to ...

mouse-pneumonectomy-model-of-compensatory-lung-growth

Research

JoVE Journal

Medicine

12.7K 次观看.  Cincinnati Children's Hospital Medical Center. Mouse pneumonectomy is a commonly employed model of compensatory lung growth. This procedure can be used in conjunction with lineage tracing or transgenic mouse models to elucidate underlying mechanisms.

视频: Mouse Pneumonectomy Model of Compensatory Lung Growth

A Simple Method of Mouse Lung Intubation

A simple procedure to intubate mice for pulmonary function measurements would have several advantages in longitudinal studies with limited numbers or expensive animal. One of the reasons that this is not done more routinely is that it is relatively difficult, despite there being several published studies that describe ways to achieve it. In this paper we demonstrate a procedure that eliminates one of the major hurdles associated with this intubation, that of visualizing the trachea during the entire time of intubation. The approach uses a 0.5 mm fiberoptic light source that serves as an introducer to direct the intubation cannula into the mouse trachea. We show that it is possible to use this procedure to measure lung mechanics in individual mice over a time course of at least several weeks. The technique can be set up with relatively little expense and expertise, and it can be routinely accomplished with relatively little training. This should make it possible for any laboratory to routinely carry out this intubation, thereby allowing longitudinal studies in individual mice, thereby minimizing the number of mice needed and increasing the statistical power by using each mouse as its own control.

This paper describes a striaghforward and efficient method of intubating mice for pulmonary function measurements or pulmonary instillation, that allows the mice to recover and be studied at later times. The procedure involves an inexpensive fiberoptic light source that directly illuminates the trachea.

一个简单的方法小鼠肺插管

A  simple procedure to intubate mice for pulmonary function measurements would  have several advantages in longitudinal studies with limited numbers or  ...

科研

医学

Diagnostic Ultrasound Imaging of Mouse Diaphragm Function

Function analysis of rodent respiratory skeletal muscles, particularly the diaphragm, is commonly performed by isolating muscle strips using invasive surgical procedures. Although this is an effective method of assessing in vitro diaphragm activity, it involves non-survival surgery. The application of non-invasive ultrasound imaging as an in vivo procedure is beneficial since it not only reduces the number of animals sacrificed, but is also suitable for monitoring disease progression in live mice. Thus, our ultrasound imaging method may likely assist in the development of novel therapies that alleviate muscle injury induced by various respiratory diseases. Particularly, in clinical diagnoses of obstructive lung diseases, ultrasound imaging has the potential to be used in conjunction with other standard tests to detect the early onset of diaphragm muscle fatigue. In the current protocol, we describe how to accurately evaluate diaphragm contractility in a mouse model using a diagnostic ultrasound imaging technique.

Diagnostic ultrasound imaging has proven to be effective in diagnosing various respiratory diseases in human and animal subjects. We demonstrate a comprehensive ultrasound protocol utilized by Dr. Zuo's lab to analyze diaphragm kinetics specifically in mouse models. This is also a non-invasive research technique which can provide quantitative information on mouse respiratory muscle function.

小鼠膈肌功能诊断超声成像

Function analysis of rodent respiratory skeletal muscles, particularly the diaphragm, is commonly performed by isolating muscle strips using invasive ...

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 ...

Implantation of Fibrin Gel on Mouse Lung to Study Lung-specific Angiogenesis

Recent significant advances in stem cell research and bioengineering techniques have made great progress in utilizing biomaterials to regenerate and repair damage in simple tissues in the orthopedic and periodontal fields. However, attempts to regenerate the structures and functions of more complex three-dimensional (3D) organs such as lungs have not been very successful because the biological processes of organ regeneration have not been well explored. It is becoming clear that angiogenesis, the formation of new blood vessels, plays key roles in organ regeneration. Newly formed vasculatures not only deliver oxygen, nutrients and various cell components that are required for organ regeneration but also provide instructive signals to the regenerating local tissues. Therefore, to successfully regenerate lungs in an adult, it is necessary to recapitulate the lung-specific microenvironments in which angiogenesis drives regeneration of local lung tissues. Although conventional in vivo angiogenesis assays, such as subcutaneous implantation of extracellular matrix (ECM)-rich hydrogels (e.g., fibrin or collagen gels or Matrigel - ECM protein mixture secreted by Engelbreth-Holm-Swarm mouse sarcoma cells), are extensively utilized to explore the general mechanisms of angiogenesis, lung-specific angiogenesis has not been well characterized because methods for orthotopic implantation of biomaterials in the lung have not been well established. The goal of this protocol is to introduce a unique method to implant fibrin gel on the lung surface of living adult mouse, allowing for the successful recapitulation of host lung-derived angiogenesis inside the gel. This approach enables researchers to explore the mechanisms by which the lung-specific microenvironment controls angiogenesis and alveolar regeneration in both normal and pathological conditions. Since implanted biomaterials release and supply physical and chemical signals to adjacent lung tissues, implantation of these biomaterials on diseased lung can potentially normalize the adjacent diseased tissues, enabling researchers to develop new therapeutic approaches for various types of lung diseases.

Recapitulation of the organ-specific microenvironment, which stimulates local angiogenesis, is indispensable for successful regeneration of damaged tissues. This report demonstrates a novel method to implant fibrin gels on the lung surface of living mouse in order to explore how the lung-specific microenvironment modulates angiogenesis and alveolar regeneration in adult mouse.

纤维蛋白胶对小鼠肺移植研究肺特异性血管生成

Recent significant advances in stem cell research and bioengineering techniques have made great progress in utilizing biomaterials to regenerate and ...

Method of Isolated Ex Vivo Lung Perfusion in a Rat Model: Lessons Learned from Developing a Rat EVLP Program

The number of acceptable donor lungs available for lung transplantation is severely limited due to poor quality. Ex-Vivo Lung Perfusion (EVLP) has allowed lung transplantation in humans to become more readily available by enabling the ability to assess organs and expand the donor pool. As this technology expands and improves, the ability to potentially evaluate and improve the quality of substandard lungs prior to transplant is a critical need. In order to more rigorously evaluate these approaches, a reproducible animal model needs to be established that would allow for testing of improved techniques and management of the donated lungs as well as to the lung-transplant recipient. In addition, an EVLP animal model of associated pathologies, e.g., ventilation induced lung injury (VILI), would provide a novel method to evaluate treatments for these pathologies. Here, we describe the development of a rat EVLP lung program and refinements to this method that allow for a reproducible model for future expansion. We also describe the application of this EVLP system to model VILI in rat lungs. The goal is to provide the research community with key information and &#8220;pearls of wisdom&#8221;/techniques that arose from trial and error and are critical to establishing an EVLP system that is robust and reproducible.

Method of Isolated Ex Vivo Lung Perfusion in a Rat Model: Lessons Learned from Developing a Rat EVLP Program

Ex-Vivo Lung Perfusion (EVLP) has allowed lung transplantation in humans to become more readily available by enabling the ability to assess organs and expand the donor pool. Here, we describe the development of a rat EVLP program and refinements that allow for a reproducible model for future expansion.

的隔离方法<em&gt;离体</em&gt;肺灌注的大鼠模型:教训制定鼠EVLP课程中学

The number of acceptable donor lungs available for lung transplantation is severely limited due to poor quality. Ex-Vivo Lung Perfusion (EVLP) has allowed ...

Real-time X-ray Imaging of Lung Fluid Volumes in Neonatal Mouse Lung

At birth, the lung undergoes a profound phenotypic switch from secretion to absorption, which allows for adaptation to breathing independently. Promoting and sustaining this phenotype is critically important in normal alveolar growth and gas exchange throughout life. Several in vitro studies have characterized the role of key regulatory proteins, signaling molecules, and steroid hormones that can influence the rate of lung fluid clearance. However, in vivo examinations must be performed to evaluate whether these regulatory factors play important physiological roles in regulating perinatal lung liquid absorption. As such, the utilization of real time X-ray imaging to determine perinatal lung fluid clearance, or pulmonary edema, represents a technological advancement in the field. Herein, we explain and illustrate an approach to assess the rate of alveolar lung fluid clearance and alveolar flooding in C57BL/6 mice at post natal day 10 using X-ray imaging and analysis. Successful implementation of this protocol requires prior approval from institutional animal care and use committees (IACUC), an in vivo small animal X-ray imaging system, and compatible molecular imaging software.

We present a protocol to assess the rate of alveolar fluid clearance or pulmonary edema in neonatal mouse lung using X-ray imaging technology.

在新生小鼠肺肺流体体积的实时X射线成像

At birth, the lung undergoes a profound phenotypic switch from secretion to absorption, which allows for adaptation to breathing independently. Promoting ...

Apical Resection Mouse Model to Study Early Mammalian Heart Regeneration

Cardiovascular disease plagues the whole world due to intensive lifestyle changes. Heart regeneration holds great promise for repairing and restoring cardiomyocytes lost due to injury and disease. In contrast to the robust cardiac regeneration of certain lower vertebrates, adult mammalian hearts typically show minimal capacity for heart regeneration and repair. However, recent studies have sparked considerable scientific interest with the finding that, between postnatal day 1 to 7 (P1 to P7), the neonatal mouse heart retains significant regenerative potential after apical resection (i.e., surgical amputation and exposure of left ventricular apex). One major controversy over this finding might be due to the diverse surgery-related procedures used in efforts to replicate or expand upon this important finding. These instructions dynamically present the materials and methodology for apical resection in a mouse model. The salient steps of this rodent survival surgery involve hypothermia anesthesia, thoracotomy, surgical amputation of heart ventricular apex, and suture and recovery of mice. The approach described could expand the application of the apical resection mouse model for cardiovascular research.

A step-by-step video protocol of apical resection is demonstrated in this study. Apical resection is a recently highlighted surgical approach in mammalian heart regeneration research. This study may promote the application of apical resection as a standard methodology in research into the mechanism underlying heart regeneration.

根尖切除小鼠模型来研究早期哺乳动物心脏再生

Cardiovascular disease plagues the whole world due to intensive lifestyle changes. Heart regeneration holds great promise for repairing and restoring ...

Open Tracheostomy Gastric Acid Aspiration Murine Model of Acute Lung Injury Results in Maximal Acute Nonlethal Lung Injury

Acid pneumonitis is a major cause of sterile acute lung injury (ALI) in humans. Acid pneumonitis spans the clinical spectrum from asymptomatic to acute respiratory distress syndrome (ARDS), characterized by neutrophilic alveolitis, and injury to both alveolar epithelium and vascular endothelium. Clinically, ARDS is defined by acute onset of hypoxemia, bilateral patchy pulmonary infiltrates and non-cardiogenic pulmonary edema. Human studies have provided us with valuable information about the physiological and inflammatory changes in the lung caused by ARDS, which has led to various hypotheses about the underling mechanisms. Unfortunately, difficulties determining the etiology of ARDS, as well as a wide range of pathophysiology have resulted in a lack of critical information that could be useful in developing therapeutic strategies.
Translational animal models are valuable when their pathogenesis and pathophysiology accurately reproduce a concept proven in both in vitro and clinical settings. Although large animal models (e.g., sheep) share characteristics of the anatomy of human trachea-bronchial tree, murine models provide a host of other advantages including: low cost; short reproductive cycle lending itself to greater data acquisition; a well understood immunologic system; and a well characterized genome leading to the availability of a variety of gene deletion and transgenic strains. A robust model of low pH induced ARDS requires a murine ALI that targets mainly the alveolar epithelium, secondarily the vascular endothelium, as well as the small airways leading to the alveoli. Furthermore, a reproducible injury with wide differences between different injurious and non-injurious insults is important.
The murine gastric acid aspiration model presented here using hydrochloric acid employs an open tracheostomy and recreates a pathogenic scenario that reproduces the low pH pneumonitis injury in humans. Additionally, this model can be used to examine interaction of a low pH insult with other pulmonary injurious entities (e.g., food particles, pathogenic bacteria).

This protocol induces acute lung injury in a mouse that has close fidelity to the pathogenesis of acid pneumonitis observed in humans. We generate a maximal acute nonlethal low pH lung injury and account for differences in rodent-human anatomic respiratory structure using an open tracheostomy coupled with circumferential pressure release.

急性肺损伤开放气管切开胃酸吸入小鼠模型结果在急性最大非致命性肺损伤

Acid pneumonitis is a major cause of sterile acute lung injury (ALI) in humans. Acid pneumonitis spans the clinical spectrum from asymptomatic to acute ...

Integrated Compensatory Responses in a Human Model of Hemorrhage

Hemorrhage is the leading cause of trauma-related deaths, partly because early diagnosis of the severity of blood loss is difficult. Assessment of hemorrhaging patients is difficult because current clinical tools provide measures of vital signs that remain stable during the early stages of bleeding due to compensatory mechanisms. Consequently, there is a need to understand and measure the total integration of mechanisms that compensate for reduced circulating blood volume and how they change during ongoing progressive hemorrhage. The body&#39;s reserve to compensate for reduced circulating blood volume is called the &#39;compensatory reserve&#39;. The compensatory reserve can be accurately evaluated with real-time measurements of changes in the features of the arterial waveform measured with the use of a high-powered computer. Lower Body Negative Pressure (LBNP) has been shown to simulate many of the physiological responses in humans associated with hemorrhage, and is used to study the compensatory response to hemorrhage. The purpose of this study is to demonstrate how compensatory reserve is assessed during progressive reductions in central blood volume with LBNP as a simulation of hemorrhage.

The purpose of this protocol is to demonstrate the techniques for measuring compensatory responses to reduced central blood volume using lower body negative pressure as a noninvasive experimental model of human hemorrhage which can be used to quantify the total integration of compensatory mechanisms to blood volume deficit in humans.

在出血的人体模型集成的代偿反应

Hemorrhage is the leading cause of trauma-related deaths, partly because early diagnosis of the severity of blood loss is difficult. Assessment of ...

Echocardiographic Measurement of Right Ventricular Diastolic Parameters in Mouse

Diastolic dysfunction is a prominent feature of right ventricular (RV) remodeling associated with conditions of pressure overload. However, the RV diastolic function is rarely quantified in experimental studies. This might be due to technical difficulties in the visualization of the RV in the apical four-chamber view in rodents. Here we describe two positions facilitating the visualization of the apical four-chamber view in mice to assess the RV diastolic function.
The apical four-chamber view is enabled by tilting the mouse fixation platform to the left and caudally (LeCa) or to the right and cranially (RiCr). Both positions provide images of comparable quality. The results of the RV diastolic function obtained from two positions are not significantly different. Both positions are comparably easy to perform. This protocol can be incorporated into published protocols and enables detailed investigations of the RV function.

Here we describe and compare two positions for obtaining the apical four-chamber view in mice. These positions enable the quantification of the right ventricular function, provide comparable results, and can be used interchangeably.