Name: direct intrabronchial administration to improve the selective agent deposition within the mouse lung
Uploaded: 2019-05-20T14:00:00
Description: Watch this Scientific Journal Video about Direct Intrabronchial Administration to Improve the Selective Agent Deposition Within the Mouse Lung at JoVE.com

这项工作由NHLBI向E.P.S.提供HL125371赠款和国防部(CDMRP)赠款W81XWH-17-1-0051给Y.Y.Y.

所有动物协议都已获得科罗拉多大学丹佛机构动物护理和使用委员会(IACUC)的批准。下面描述的所有程序(第 4-7 节)都使用雄性和雌性 C57BL/6 小鼠进行了优化。这种方法已经验证使用小鼠范围从19-40克的体重。
1. 创建IB管理平台
<ol><li>将书尾从基底翼和站立翼之间的原始 90° 角度弯曲到 70°(图 2A)。</li>
	<li>在金属书底立翼的中端钻一个孔(?...

<ol>
	<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 (3), 984-987 (2009).</li><li>Thomas, J. L., 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=Endotracheal+intubation+in+mice+via+direct+laryngoscopy+using+an+otoscope.">Endotracheal intubation in mice via direct laryngoscopy using an otoscope.</a> Journal of Visualized Experiments. (86), (2014).</li><li>Vandivort, T. C., An, D., Parks, W. C. <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+Method+for+Rapid+Intubation+of+the+Trachea+in+Mice.">An Improved Method for Rapid Intubation of the Trachea in Mice.</a> Journal of Visualized Experiments. (108), 53771 (2016).</li><li>McGovern, T. K., Robichaud, A., Fereydoonzad, L., Schuessler, T. F., Martin, J. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Evaluation+of+respiratory+system+mechanics+in+mice+using+the+forced+oscillation+technique.">Evaluation of respiratory system mechanics in mice using the forced oscillation technique.</a> Journal of Visualized Experiments. (75), e50172 (2013).</li><li>Halbower, A. C., Mason, R. J., Abman, S. H., Tuder, R. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Agarose+infiltration+improves+morphology+of+cryostat+sections+of+lung.">Agarose infiltration improves morphology of cryostat sections of lung.</a> Laboratory Investigation. 71 (1), 149-153 (1994).</li><li>Thrall, R. S., McCormick, J. R., Jack, R. M., McReynolds, R. A., Ward, P. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Bleomycin-induced+pulmonary+fibrosis+in+the+rat:+inhibition+by+indomethacin.">Bleomycin-induced pulmonary fibrosis in the rat: inhibition by indomethacin.</a> American Journal of Pathology. 95 (1), 117-130 (1979).</li><li>Matute-Bello, G., Frevert, C. W., Martin, T. R. <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+acute+lung+injury.">Animal models of acute lung injury.</a> American Journal of Physiology Lung Cellular and Molecular Physiology. 295 (3), L379-L399 (2008).</li><li>Del Sorbo, L., 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=Intratracheal+Administration+of+Small+Interfering+RNA+Targeting+Fas+Reduces+Lung+Ischemia-Reperfusion+Injury.">Intratracheal Administration of Small Interfering RNA Targeting Fas Reduces Lung Ischemia-Reperfusion Injury.</a> Criticial Care Medicine. 44 (8), e604-e613 (2016).</li><li>McLemore, T. L., 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=Novel+intrapulmonary+model+for+orthotopic+propagation+of+human+lung+cancers+in+athymic+nude+mice.">Novel intrapulmonary model for orthotopic propagation of human lung cancers in athymic nude mice.</a> Cancer Research. 47 (19), 5132-5140 (1987).</li><li>Vertrees, R. 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=Development+of+a+human+to+murine+orthotopic+xenotransplanted+lung+cancer+model.">Development of a human to murine orthotopic xenotransplanted lung cancer model.</a> Journal of Investigative Surgery. 13 (6), 349-358 (2000).</li></ol>

肺损伤在啮齿类动物中采用IT管理,如BLM6。肺损伤是典型的模型。然而,这种IT管理只会导致零碎的伤害,反映了这种方法7的肺分娩的非针对性。模拟肺损伤的这些局限性是尝试提供非有害实验性药物(如药物、siRNA 或细胞疗法)的 IT 时面临的指导性挑战。
在本报告中,我们描述了对实验代理的直接IB管理。与传统的 IT 管理方法不同,此方法具有两...

小鼠实验剂的直接肺给给术可用于研究肺免疫反应、急性肺损伤和肺纤维化。直接肺管理通常通过内切术 (IT) 灌输执行,如前面描述的1,2,3。然而,这种方法是非选择性的,以非靶向和经常不对称的方式影响两个肺。 肺损伤的实验建模可能受益于选择性地瞄准一个特定肺的能力,允许使用反向肺作为对照。相反,人类扩散性肺病的精确建模得益于实验剂对称分布到双边肺气肿。
本报告的总体目标是描述一种选择性地将实验剂输送到小鼠左肺或右肺的方法(图1)。这种支管内(IB)管理方法允许对小鼠肺进行单方面治疗,并可轻松加以调整,以确保将制剂平等交付给双边主干支气管。通过使用IB管理将大剂量的实验剂输送到较大的右肺,将更小的体积输送到较小的左肺(即剂量调整的IB给药),本报告证明的是肺的同质性的改善提供实验剂,优化小鼠扩散性肺损伤模型。因此,这份报告对于寻求单方面给小鼠施用实验剂或改善两个肺部药物沉积的对称性的调查者可能具有价值。

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		<tr height="21">
			<td height="21" style="height:21px;width:280px;">22 G shielded IV Catheter</td>
			<td style="width:139px;">BD</td>
			<td style="width:140px;">381423</td>
			<td style="width:180px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:280px;">Bleomycin</td>
			<td style="width:139px;">Enzo life sciences</td>
			<td style="width:140px;">BML-AP302-0010</td>
			<td></td>
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			<td height="42" style="height:42px;width:280px;">Compact Mini rodent anesthesia machine&#160;</td>
			<td style="width:139px;">DRE Veterinary</td>
			<td style="width:140px;">9280</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:280px;">Evans blue dye</td>
			<td style="width:139px;">Sigma-Aldrich</td>
			<td style="width:140px;">E2129</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:280px;">FITC-dextran</td>
			<td style="width:139px;">Sigma-Aldrich</td>
			<td style="width:140px;">FD150</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:280px;">Isoflurane</td>
			<td style="width:139px;">Piramal Critical Care</td>
			<td style="width:140px;">NDC&#160;</td>
			<td></td>
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		<tr height="42">
			<td height="42" style="height:42px;width:280px;">LED-30W Fiber Optic Dual Gooseneck Lights Microscope Illuminator</td>
			<td style="width:139px;">AmScope</td>
			<td style="width:140px;">LED-30W</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Low temperature cautery with fine tip&#160;</td>
			<td style="width:139px;">Bovie</td>
			<td style="width:140px;">AA02</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:280px;">Precisionglide needle, 18G x 1&#34;</td>
			<td style="width:139px;">BD</td>
			<td style="width:140px;">305195</td>
			<td>Beveled tip, 12 mm in length&#160;</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:280px;">Xylazine</td>
			<td style="width:139px;">AKORN</td>
			<td style="width:140px;">NDC 59399-110-20</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:280px;">Zatamine</td>
			<td>VetOne</td>
			<td style="width:140px;">NDC 13985-702-10&#160;</td>
			<td>Ketamine</td>
		</tr>
	</tbody>
</table>

direct intrabronchial administration to improve the selective agent deposition within the mouse lung

实验性制剂的内切内(IT)管理是扩散性肺病(如博霉素引起的肺纤维化)的鼠模型中必不可少的技术。 然而,在远端小鼠肺中,内切开剂的分布往往是不对称的,小鼠的左肺中肺气肿浓度增加。 本报告中描述了一种新的肺内(IB)方法,用于使活小鼠的左肺和/或右肺不手术。 还演示了如何利用这种方法选择性地将试剂施用到一个肺或调整(通过剂量调整的IB交付),以改善实验剂肺输送的左右对称性,从而改进扩散模型肺病,如博洛霉素引起的肺纤维化。

选择性IB插管针对特定的叶(右肺)或巴勒拉段(左肺)。
IB给右肺的EBD给分部,如第5.1节所述。实验完成后,给小鼠注射致命剂量的腹内氯胺酮/木氨酸,并采集肺部以证明EBD分布(图4A,右图)。肺毛外观表明,90%的尝试可以使右肺的小后叶,而10%的尝试针对下等叶。据推测,这些叶的小体积解释了小鼠在远?...

Watch this Scientific Journal Video about Direct Intrabronchial Administration to Improve the Selective Agent Deposition Within the Mouse Lung at JoVE.com

Direct Intrabronchial Administration to Improve the Selective Agent Deposition Within the Mouse Lung

小鼠实验剂的内切管 (IT) 给给通常会导致对称输送到远端肺部。 在这份报告中,我们描述了一种直接的肺内(IB)方法,用于对活体小鼠的每项肺进行非手术处理。 这种方法可用于选择性地将制剂施用到一个肺上,或者可以调整以改善对称剂向两个肺的输送。

直接注射内管理,改善小鼠肺内的选择性代理沉积

Intratracheal (IT) administration of experimental agents is an essential technique in murine models of diffuse lung diseases, such as bleomycin-induced ...

为了模拟小鼠的肺病，实验药剂通常通过腹内注射进行。然而，这种方法不能有选择地瞄准一个肺，而且肺之间的分布可能是不对称的。通过麻醉小鼠单个肺，我们可以单方面管理实验剂。通过修改，这种方法可以双边使用，改善两个肺沉积的对称性。导管制备，包括抛光、轻微扬格和标记是成功的关键要素。当然，练习这种技术也是必要的。演示这个程序的将是我实验室的专业研究助理廖素梅。在确认对手趾捏没有反应后，从悬覆缝合线上的切口上悬挂麻醉剂，8到10周大，C57-Black/6 鼠标以上挂缝合线。用两到三块挂钩将鼠标松散地固定起来，并环住磁带，避免通风限制，并打开发光二极管光纤照明器。将操作员放在平台后面，将鼠标对台。将照明器的鹅颈定向到距离鼠标两到三厘米的距离，以便它通过皮肤照亮喉部区域。使用主导手的无菌钳子，将舌头从口腔中抽出。使用非主要手和无菌抑制剂压平舌头根部，广泛暴露口咽。释放钳子后，使用主导手通过口腔将延长的导管插入气管。通过观察注射器中的气泡是否随每次呼吸上下移动来确认放置位置。对于右肺的选择性侧杆，在切内插管后，旋转塑料板30度，并引导导管轮毂平行于小鼠中线到适当的重量深度，如表中所示。使用凝胶加载尖端，将 20 微升 0.3% Evans 蓝色染料送入导管，然后由玻璃滴管将 1 到 2 个 100 微升的空气等分。然后，取回导管，将鼠标保持30秒的位置，然后将动物放在加热毯上，进行监测，直到完全恢复。对于左肺的选择性腰椎，在切内插管后，旋转塑料板零下74度，轻轻推进导管到左主管，同时将适度的压力向下90度向下，朝向书本。在左下肺段进行干燥后，旋转塑料板零下30度，并交付40微升0.3%的Evas蓝色染料与凝胶加载尖端。然后，通过玻璃滴管提供一到两个100至300微升的空气等等，并提取导管，让鼠标恢复，如证明。对于右整个肺给药，在内切内插管后，旋转塑料板30度，并引导导管平行于小鼠中线，达到右侧近端腰椎间管的深。确认 tachypnea 标志的外观，并旋转鼠标零下 74 度，以便为代理交付提供重力辅助。将导管撤回到与右主管支气管起飞相对应的位置，并确保导管的斜面朝下。将 30 微升 0.3% 的 Evans 蓝色染料送到右肺，然后向右肺提供 1 到 2 个 100 到 300 微升的空气等分。然后，收回导管，让鼠标恢复。对于左整个肺给氏，在血管内弯曲后，旋转塑料板减去74度，并轻轻地将导管推进到左主管支气管，同时向下和向书本施加适度的压力为90度。确认无 tachypnea 符号，并旋转鼠标 86 度，以允许重力辅助代理管理。将导管提取到左侧主干支气管，然后旋转导管的斜面朝下。在左肺提供 30 微升 0.3% Evans 蓝色染料，并带凝胶加载尖端，然后提供 1 到 2 个 100 到 300 微升的空气等同。然后，收回导管，让鼠标恢复，如证明。通过双边交付的菲特克-德斯特兰通过中腹内和对称的支气管内给方导致不对称的肺腹质 FITC 荧光与更大的相对浓度注意到在左肺。这表明，在切内给舍后，实验剂的不对称肺传递不是这些制剂不对称地呈现给每个主茎支气管的结果，而是由肺重量和质量的差异稀释了同等主茎的输送。剂量调整的支气管内给药改善了肺腹体剂输送的对称性。然而，尽管这种修正，在右肺的不同叶内仍然观察到持续的异质性。在腹腔内出血霉素引起的纤维性肺损伤21天后，炎症主要观察在左肺，在右肺损伤较少。相反，剂量调整的支气管内布洛霉素给药导致左肺损伤略有减少，右肺损伤增加，其中叶和上叶损伤明显。在整个肺内给普过程中，不要忘记提取导管，将导管的斜面朝下。

Research

JoVE Journal

Medicine

High-frequency High-resolution Echocardiography: First Evidence on Non-invasive Repeated Measure of Myocardial Strain, Contractility, and Mitral Regurgitation in the Ischemia-reperfused Murine Heart

高频超声心动图：非侵入性的反复测量小鼠心脏缺血再灌注心肌劳损，收缩力，二尖瓣关闭不全的第一个证据

Ischemia-reperfusion (IR) was surgically performed in murine hearts which were then subjected to repeated imaging to monitor temporal changes in ...

科研

医学

Bronchial Thermoplasty:  A Novel Therapeutic Approach to Severe Asthma

支气管Thermoplasty：严重哮喘的一种新的治疗方法

Bronchial thermoplasty is a non-drug procedure for severe persistent asthma that delivers thermal energy to the airway wall in a precisely controlled ...

Noninvasive Intratracheal Intubation to Study the Pathology and Physiology of Mouse Lung

无创性气管内插管，以研究小鼠肺的病理生理

The use of a model that mimics the condition of lung diseases in humans is critical for studying the pathophysiology and/or etiology of a particular ...

Diagnostic Ultrasound Imaging of Mouse Diaphragm 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

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 ...

The Bovine Lung in Biomedical Research: Visually Guided Bronchoscopy, Intrabronchial Inoculation and In Vivo Sampling Techniques

The Bovine Lung in Biomedical Research: Visually Guided Bronchoscopy, Intrabronchial Inoculation and In Vivo Sampling Techniques

牛龙在生物医学研究：视觉引导支气管镜，支气管内接种和<em&gt;在体内</em&gt;抽样技术

There is an ongoing search for alternative animal models in research of respiratory medicine. Depending on the goal of the research, large animals as ...

Visualization of Streptococcus pneumoniae within Cardiac Microlesions and Subsequent Cardiac Remodeling

Visualization of Streptococcus pneumoniae within Cardiac Microlesions and Subsequent Cardiac Remodeling

可视化<em&gt;肺炎链球菌</em&gt;在心脏Microlesions和随后的心脏重构

During bacteremia Streptococcus pneumoniae can translocate across the vascular endothelium into the myocardium and form discrete bacteria-filled ...

Design, Fabrication, and Administration of the Hand Active Sensation Test (HASTe)

Design, Fabrication, and Administration of the Hand Active Sensation Test HASTe

设计，制造管理的手，和活动感觉测试（急速）

The concept of personalizing neurologic rehabilitation, based on individual impairments, has experienced a recent surge. In parallel, the number of ...

2-Methacryloyloxyethyl Phosphorylcholine Polymer Treatment of Complete Dentures to Inhibit Denture Plaque Deposition

全口义齿的2-甲基丙烯酰磷酰胆碱聚合物处理，以抑制义齿斑块沉积

Removable dentures made of poly (methyl methacrylate) (PMMA) are prone to bacterial adherence and dental plaque formation, which is called denture plaque. ...

Quantitative Mapping of Specific Ventilation in the Human Lung using Proton Magnetic Resonance Imaging and Oxygen as a Contrast Agent

以质子磁共振成像和氧为造影剂对人肺特异性通气进行定量定位

Specific ventilation imaging (SVI) is a functional magnetic resonance imaging technique capable of quantifying specific ventilation &#8213; the ratio of ...