Name: brain death induction in mice using intra-arterial blood pressure monitoring and ventilation via tracheostomy
Uploaded: 2020-04-17T14:00:00
Description: Watch this Scientific Journal Video about Brain Death Induction in Mice Using Intra-Arterial Blood Pressure Monitoring and Ventilation via Tracheostomy at JoVE.com

动物实验是按照国家医学研究学会制定的《动物实验室护理原则和国家科学院编制、国家卫生研究院出版的实验室动物护理和使用指南》（NIH出版物第86-23号，1985年修订版）进行的。所有实验均获得奥地利教育、科学和文化部的批准（BMWF-66.011/0071-II/3b/2012）。
1. 动脉导管化
<ol><li>根据当地惯例（如丁丙诺啡），用内腹注射氯胺酮和西拉辛10和镇痛剂麻醉小?...

<ol>
	<li>Hart, 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=OPTN/SRTR+2017+Annual+Data+Report:+Kidney.">OPTN/SRTR 2017 Annual Data Report: Kidney.</a> American Journal of Transplantation. 19 (Suppl 2), 19 (2019).</li><li>The Quality Standards Subcommittee of the American Academy of Neurology. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Practice+parameters+for+determining+brain+death+in+adults+(summary+statement).">Practice parameters for determining brain death in adults (summary statement).</a> Neurology. 45 (5), 1012-1014 (1995).</li><li>Terasaki, P. I., Cecka, J. M., Gjertson, D. W., Takemoto, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=High+survival+rates+of+kidney+transplants+from+spousal+and+living+unrelated+donors.">High survival rates of kidney transplants from spousal and living unrelated donors.</a> New England Journal Medicine. 333 (6), 333-336 (1995).</li><li>Pratschke, J., Neuhaus, P., Tullius, S. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=What+can+be+learned+from+brain-death+models?.">What can be learned from brain-death models?.</a> Transplant International. 18 (1), 15-21 (2005).</li><li>Wilhelm, M. J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Activation+of+the+heart+by+donor+brain+death+accelerates+acute+rejection+after+transplantation.">Activation of the heart by donor brain death accelerates acute rejection after transplantation.</a> Circulation. 102 (19), 2426-2433 (2000).</li><li>Pomper, 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=Introducing+a+mouse+model+of+brain+death.">Introducing a mouse model of brain death.</a> Journal of Neuroscience Methods. 192 (1), 70-74 (2010).</li><li>Ritschl, P. V., 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=Donor+brain+death+leads+to+differential+immune+activation+in+solid+organs+but+does+not+accelerate+ischaemia-reperfusion+injury.">Donor brain death leads to differential immune activation in solid organs but does not accelerate ischaemia-reperfusion injury.</a> Journal of Pathology. 239 (1), 84-96 (2016).</li><li>Atkinson, 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=Donor+brain+death+exacerbates+complement-dependent+ischemia/reperfusion+injury+in+transplanted+hearts.">Donor brain death exacerbates complement-dependent ischemia/reperfusion injury in transplanted hearts.</a> Circulation. 127 (12), 1290-1299 (2013).</li><li>Oberhuber, R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Treatment+with+tetrahydrobiopterin+overcomes+brain+death-associated+injury+in+a+murine+model+of+pancreas+transplantation.">Treatment with tetrahydrobiopterin overcomes brain death-associated injury in a murine model of pancreas transplantation.</a> American Journal of Transplantation. 15 (11), 2865-2876 (2015).</li><li>Floerchinger, 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=Inflammatory+immune+responses+in+a+reproducible+mouse+brain+death+model.">Inflammatory immune responses in a reproducible mouse brain death model.</a> Transplant Immunology. 27 (1), 25-29 (2012).</li><li>Steen, P. A., Milde, J. H., Michenfelder, J. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=No+barbiturate+protection+in+a+dog+model+of+complete+cerebral+ischemia.">No barbiturate protection in a dog model of complete cerebral ischemia.</a> Annals of Neurology. 5 (4), 343-349 (1979).</li><li>Cooper, D. K., Novitzky, D., Wicomb, W. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+pathophysiological+effects+of+brain+death+on+potential+donor+organs,+with+particular+reference+to+the+heart.">The pathophysiological effects of brain death on potential donor organs, with particular reference to the heart.</a> Annals of the Royal College of Surgeons of England. 71 (4), 261-266 (1989).</li><li>Herijgers, P., Leunens, V., Tjandra-Maga, T. B., Mubagwa, K., Flameng, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Changes+in+organ+perfusion+after+brain+death+in+the+rat+and+its+relation+to+circulating+catecholamines.">Changes in organ perfusion after brain death in the rat and its relation to circulating catecholamines.</a> Transplantation. 62 (3), 330-335 (1996).</li></ol>

BD是多器官捐赠者异体移植质量的危险因素，需要大量的病理生理变化，这些变化只能使用体内模型进行充分评估。血动力学变化、细胞因子风暴、荷尔蒙变化及其对器官移植质量和生存的最终影响不能在体外分析。大多数基础移植和免疫学研究都依赖于复杂的诊断工具，这些工具仅在小鼠模型中广泛提供。小鼠模型已经导致在BD相关研究77，8，9<sup clas...

移植是目前治疗终末期器官衰竭的唯一治疗方法。到目前为止，脑死亡（BD）患者一直是器官捐献的主要来源，尽管活体捐献和循环后死亡后捐赠是有价值的选择1。BD的定义是一个不可逆转的昏迷（与已知的原因），没有脑干反射和呼吸暂停2。不幸的是，BD器官在长期移植生存中表现出低劣的结果，而人类白细胞抗原（HLA）不匹配和冷缺血时间3。同时，对这种抗原无关危险因素进行了深入的研究，结果产生了BD引起的病理生理变化的三个主要方面：血液动力学、荷尔蒙和炎症4。
迄今为止，啮齿动物的BD实验模型大多使用大鼠进行。为了更深入地了解BD之后固体器官的免疫后果，我们旨在建立BD的鼠模型，因为目前只有小鼠模型才能对遗传或免疫因素进行全面调查。在此背景下，鼠标系统提供了种类更丰富的分析工具。
此处描述的BD感应原理基于颅骨下插入的气球导管的膨胀引起的颅内压力增加。增加的颅内压力模仿BD的生理机制，通过阻断脑脑、小脑和脑干的灌注55，6。6为了保证周围器官充分灌注，在手术过程中必须测量血压。同时用于此目的的导管用于盐水管理，以便通过液体替代来稳定血压。由于BD伴随着自发呼吸的停止，必须确保充足的通风。电热毯能维持生理核心体温。
综上所述，该模型将能深入研究BD诱发损伤的影响、白细胞迁移7、恭维激活8、缺血再灌注损伤9等因素。

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Arterial catheter (BD Neoflon 26G)</td>
			<td>BD</td>
			<td>391349</td>
			<td></td>
		</tr>
		<tr>
			<td>Blood Pressure Transducers (APT300)</td>
			<td>Harvard Apparatus Inc.</td>
			<td>73-3862</td>
			<td></td>
		</tr>
		<tr>
			<td>Fogarty Arterial Embolectomy Catheter N&#176; 3</td>
			<td>Edwards Lifesciences Corporation</td>
			<td>120403F</td>
			<td></td>
		</tr>
		<tr>
			<td>Forceps</td>
			<td>FST</td>
			<td>11271-30</td>
			<td></td>
		</tr>
		<tr>
			<td>Homeothermic Blanket Systems with Flexible Probe</td>
			<td>Harvard Apparatus Inc.</td>
			<td>55-7020</td>
			<td></td>
		</tr>
		<tr>
			<td>Ketansol</td>
			<td>Graeub</td>
			<td>6680110</td>
			<td></td>
		</tr>
		<tr>
			<td>Micro scissor</td>
			<td>FST</td>
			<td>15018-10</td>
			<td></td>
		</tr>
		<tr>
			<td>Needle holder</td>
			<td>FST</td>
			<td>12060-02</td>
			<td></td>
		</tr>
		<tr>
			<td>Prolene 5-0</td>
			<td>Ethicon</td>
			<td>8698H</td>
			<td></td>
		</tr>
		<tr>
			<td>Pump 11 Elite Infusion Only Single</td>
			<td>Harvard Apparatus Inc.</td>
			<td>70-4500</td>
			<td></td>
		</tr>
		<tr>
			<td>Scissor</td>
			<td>FST</td>
			<td>14075-11</td>
			<td></td>
		</tr>
		<tr>
			<td>Stereotactic microscope</td>
			<td>Olympus</td>
			<td>SZX7</td>
			<td></td>
		</tr>
		<tr>
			<td>Transpore Tape</td>
			<td>3M</td>
			<td>1527-1</td>
			<td></td>
		</tr>
		<tr>
			<td>Underpads</td>
			<td>Molinea.A</td>
			<td>274301</td>
			<td></td>
		</tr>
		<tr>
			<td>Ventilator for mice (MiniVent Model 845)</td>
			<td>Harvard Apparatus Inc.</td>
			<td>73-0043</td>
			<td></td>
		</tr>
		<tr>
			<td>Xylasol</td>
			<td>Graeub</td>
			<td>7630109</td>
			<td></td>
		</tr>
	</tbody>
</table>

brain death induction in mice using intra-arterial blood pressure monitoring and ventilation via tracheostomy

虽然活体捐献和循环后死亡捐赠为器官移植提供了替代机会，但捐献者脑死亡后捐献（BD）仍然是实体移植的主要来源。不幸的是，大脑功能的不可逆丧失已知会导致多种病理生理变化，包括血液动力学和荷尔蒙修饰，最终导致全身炎症反应。允许在体内系统地研究这些影响的模型很少。我们提出了BD诱导的鼠模型，可以帮助调查BD对异体质量的破坏性影响。通过常见的胡萝卜动脉实施动脉内血压测量，并通过气管切开术进行可靠的通气后，BD 通过气球导管稳步增加颅内压力诱导。BD诱导后四小时，可以采集器官进行分析或进一步移植程序。我们的战略使人体内的BD能够全面分析，从而深入了解固体器官移植中BD相关的影响，并有可能为优化器官预置铺平道路。

鼠BD模型成功执行超过100次，成功率超过90%。此外，心肾介入后器官移植已安全进行7项。
BD 诱导各种病理生理学变化，可能使用此模型进一步研究。如图1所示，血压显示初始高血压峰值，然后是长时间的期。为了避免低血压的有害生理影响，排除了血压长期不足的小鼠（MAP < 50 mmHg 超?...

Watch this Scientific Journal Video about Brain Death Induction in Mice Using Intra-Arterial Blood Pressure Monitoring and Ventilation via Tracheostomy at JoVE.com

Brain Death Induction in Mice Using Intra-Arterial Blood Pressure Monitoring and Ventilation via Tracheostomy

我们提出了脑死亡诱导的鼠模型，以评估其病理生理学效应对器官的影响，以及固体器官移植背景下的连续移植的影响。

使用动脉内血压监测和通气通过气管切除术的小鼠脑死亡诱导

While both living donation and donation after circulatory death provide alternative opportunities for organ transplantation, donation after donor brain ...

虽然捐赠者脑死亡后捐赠是固体器官移植的主要来源，但脑功能的不可逆转的丧失会导致病理生理变化，导致全身炎症反应。这种脑死亡诱导的穆林模型允许使用各种各样的分析工具和敲除模型来研究脑死亡调节的路径。在确认对踏板反射缺乏反应并去除上腹部毛后，动脉导管，用 70% 乙醇对裸露的皮肤进行消毒，并使用解剖剪刀在皮肤上进行中线切口。使用钳子，钝解解亚肌腺和颈部肌肉组织，并分离组织，以暴露常见的胡萝卜动脉。放置三个 8-0右侧普通胡萝卜动脉下方的丝绸连字，并在近端结字上放置一个夹子。使动脉的张力，使水流暂停，并关闭最直的连结。插入 26 量的动脉导管，通过切口颅面上的一个小预制皮肤孔。如果流明太大，无法减少回流，用所有三种缝合线固定导管，则使用单个 5-0 单丝不可吸收，将导管固定到预成型皮肤孔附近的皮肤上。用钳子挤压导管。要进行气管切除术， 用钳子钝解解前切肌肉， 并放置两个 8 - 0气管下面的丝绸连字。在两个气管卡皮之间插入通风管，以避免单方面通风，然后用两个准备好的连字固定管，然后用 6-0 单丝不吸的运行缝合线关闭皮肤，并通风鼠标的频率为每分钟 150 次，潮汐体积为 200 微升。为了在实验动物中诱发脑死亡，将鼠标安排到易发位置，用钳子握住皮肤，用手术剪刀将皮肤从头骨上取下。在左皮层上方钻一毫米的卡钳孔，用钝钳穿透头骨的最终组织桥。移除任何锋利的边缘后，插入一个充气导管，预加盐水，所有空气都已疏散。当导管完全在颅腔内时，使用注射器泵在 10 到 15 分钟内以每分钟约 0.1 毫米的速度开始充气导管。当脑死亡发生时，停止气球导管的膨胀，并在鼠标上放置加热毯以避免体温过低。脑死亡确认后，定期监测和记录血压，每30分钟注入100微升盐水，以稳定动物的血压。脑死亡四小时后，排除任何动物不跳动的心脏，并收获鼠标器官和感兴趣的组织，根据标准协议。脑死亡诱导后，血压呈现初始高血压峰值，然后是长期的低血压阶段。另一个成熟的观察是，脑死亡诱导导致免疫系统的激活。事实上，在这个模型中，经过四个小时的脑死亡后，在mRNA水平上观察到免疫标记的器官特异性上调节。在脑死亡的预定义期后，器官可以收获用于直接分析或后续器官移植。该模型将有助于深入研究脑死亡诱发损伤的影响，并且已经揭示了补充活化和白细胞迁移的新见解。

Research

JoVE Journal

Neuroscience

Monitoring Changes in the Intracellular Calcium Concentration and Synaptic Efficacy in the Mollusc Aplysia

Monitoring Changes in the Intracellular Calcium Concentration and Synaptic Efficacy in the Mollusc Aplysia

监测变化的细胞内钙离子浓度和突触效能的软体动物<em&gt;海兔</em

It has been suggested that changes in intracellular calcium mediate the induction of a number of important forms of synaptic plasticity (e.g., ...

科研

神经科学

Functional Neuroimaging Using Ultrasonic Blood-brain Barrier Disruption and Manganese-enhanced MRI

使用超声波血脑屏障破坏和锰增强MRI的脑功能成像

Although mice are the dominant model system for studying the genetic and molecular underpinnings of neuroscience, functional neuroimaging in mice remains ...

Modeling Neuronal Death and Degeneration in Mouse Primary Cerebellar Granule Neurons

小鼠小脑颗粒神经元神经元死亡和变性的模型研究

Cerebellar granule neurons (CGNs) are a commonly used neuronal model, forming an abundant homogeneous population in the cerebellum. In light of their ...

Imaging Neural Activity in the Primary Somatosensory Cortex Using Thy1-GCaMP6s Transgenic Mice

Imaging Neural Activity in the Primary Somatosensory Cortex Using Thy1-GCaMP6s Transgenic Mice

胸腺1-gmcmp6 转基因小鼠在原代体血源皮质中的成像神经活性

The mammalian brain exhibits marked symmetry across the sagittal plane. However, detailed description of neural dynamics in symmetric brain regions in ...

An In Vivo Blood-brain Barrier Permeability Assay in Mice Using Fluorescently Labeled Tracers

An In Vivo Blood-brain Barrier Permeability Assay in Mice Using Fluorescently Labeled Tracers

用荧光标记示踪剂对小鼠血脑屏障通透性的体内检测

Blood-brain barrier (BBB) is a specialized barrier that protects the brain microenvironment from toxins and pathogens in the circulation and maintains ...

Monitoring Neuronal Survival via Longitudinal Fluorescence Microscopy

通过纵向荧光显微镜监测新生儿生存

Standard cytotoxicity assays, which require the collection of lysates or fixed cells at multiple time points, have limited sensitivity and capacity to ...

Assessment of Long-term Depression Induction in Adult Cerebellar Slices

成人脑切片长期抑郁症诱导评估

Synaptic plasticity provides a mechanism for learning and memory. For cerebellar motor learning, long-term depression (LTD) of synaptic transmissions from ...

Induction of Leptomeningeal Cells Modification Via Intracisternal Injection

通过宫内注射诱导利普托门宁皮细胞修饰

The protocol outlined here describes how to safely and manually inject solutions through the cisterna magna while eliminating the risk of damage to the ...

Intra-Arterial Delivery of Neural Stem Cells to the Rat and Mouse Brain: Application to Cerebral Ischemia

神经干细胞向大鼠和小鼠大脑的动脉内传递：对脑缺血的应用

Neural stem cell (NSC) therapy is an emerging innovative treatment for stroke, traumatic brain injury and neurodegenerative disorders. As compared to ...

Induction of Complete Transection-Type Spinal Cord Injury in Mice

小鼠完全转型脊髓损伤的诱导

Spinal cord injury (SCI) largely leads to irreversible and permanent loss of function, most commonly as a result of trauma. Several treatment options, ...