我们承认以下资金来源:NIH NINDS – K08NS101122 （JB）， R01NS040337 （JK）， R01NS044370 （JK）， 弗吉尼亚大学医学院 （JB）。

所有动物研究均获得弗吉尼亚大学机构动物护理和使用委员会（IACUC）的批准。
1. 电极建筑/电缆建筑
<ol><li>使用单极绝缘不锈钢丝（0.005"裸直径，0.008"涂层）制成与母插座连接器（母插座连接器0.079）连接的电极。</li>
	<li>使用特殊的定制电缆将动物连接到放大器。<ol>
<li>将一个 4 针公连接器（公连接器 0.079"）连接到 4 通道单位增益阻抗匹配运算放大器（运算放大器）。将10K电阻连接到连接到9 V电池的电线上。未连接到运算放大器的地线充当电池的中点。</li>
		<li>将电缆的一端（AWG，0.012" 外径）连接到运算放大器，并将电缆的另一端连接到放大器。</li>
	</ol>
</li>
</ol>2. 电极植入手术
<ol><li>在向动的罩中用 4 - 5% 异氟醚麻醉幼崽（出生后第 9 天）。在手术开始之前，向幼崽注射布比卡因（0.02-0.05 mL，0.25%皮下局部浸润）。</li>
	<li>一旦动物无法移动，就转移到带有鼻锥的立体定向阶段。使用耳杆的反面，因为它很软，可以保持头部稳定。在这个年龄，幼崽没有完全发育的耳朵来使用耳杆的尖头。</li>
	<li>降低异氟醚的流量，并将其保持在2.5-3%。在整个手术过程中，请密切注意幼犬的稳定呼吸。捏住尾巴以检查疼痛反应，然后进行切口。</li>
	<li>用β-丁和酒精（3个循环的交替碘和70%乙醇）对颅骨上的切口区域进行灭菌。覆盖周围的身体部位，使切口区域可见。</li>
	<li>从眼睛上方稍微打开头皮前后，缩回约0.5厘米的皮肤。将鼠标头部重新定位在立体定位阶段，以便皮肤向外拉出暴露的头骨。</li>
	<li>使用棉签将过氧化氢涂抹在颅骨上，然后用手术刀刀片将颅骨刮干净。头骨非常柔软;刮擦时要小心。</li>
	<li>涂抹一滴（约50μL）粘合剂，并使用其涂抹器将其涂抹在暴露的颅骨周围。暴露在紫外线下40秒以固定粘合剂。</li>
	<li>使用暴露的边角作为参考来测量坐标。种植电极在海马体的CA1区域[-3.5 mm背腹（DV），±2 mm内侧-侧（ML），-1.75 mm深（D）]和双侧在顶叶皮层[-1.22 mm DV，±0.5 mm ML，-1 mm D]和小脑中的参考电极17。使用32 G针在标记区域创建一个孔。</li>
	<li>清洁颅骨表面的血液。在立体定位臂的帮助下，连接到母插座连接器的下电极进入大脑，并用牙科丙烯酸固定到位。将电极植入大脑。插座连接器耳机位于颅骨顶部，由牙科丙烯酸粘合在一起。</li>
	<li>电极固定后，皮下注射酮洛芬（5mg / kg）在肩胛间区域。将幼崽放回母亲身边。 注意:一次将一半的带有耳机的垃圾介绍给母亲，而不是一次介绍一个。这将防止母亲损坏小狗的耳机。</li>
</ol>3. 脑电图设置和记录（基线/损伤前）
<ol><li>电极植入后恢复24小时后，将每只动物置于加热（37°C）定制的Plexiglas室中进行脑电图记录。该室还将用作缺氧室。</li>
	<li>通过柔性电缆（定制运算放大器电缆）将腔室中的幼崽连接到视频脑电图监测系统。 注意:戴上耳机后，鼠标可以自由移动，并且不会表现出任何行为差异。一旦连接到电极丝上，必须在腔室系绳内调整导线，以提供适量的松弛度，以便幼犬可以在整个腔室中自由移动。</li>
	<li>使用草地放大器以 1K 增益对 1000 Hz 的 EEG 数据进行数字化处理。稍后使用软件（例如LabChart Pro）查看EEG信号（3-70 Hz之间的带通滤波器）。</li>
	<li>在断开动物进行颈动脉结扎手术之前，记录损伤前基线脑电图30分钟。</li>
</ol>4. 左颈动脉结扎
<ol><li>在向动的罩中用 4 - 5% 异氟醚麻醉幼犬（出生后第 10 天），并将它们放在水浴垫上特别布置的设置上。定位动物仰卧位并用纸胶带固定前肢。<ol>
<li>将异氟醚的流量降低至2-3%。捏住尾巴以获得疼痛反应，并在整个手术过程中监测呼吸。</li>
	</ol>
</li>
	<li>用甜菜碱和酒精（3个循环的交替碘和70%乙醇）对颈部左侧的切口区域（下颌骨和锁骨之间）进行消毒。</li>
	<li>使用微剪刀在颈部左侧做一个约1厘米长的切口。使用解剖显微镜，小心地缩回皮下组织和皮肤以暴露颈动脉。小心识别迷走神经（横向延伸到动脉），并小心地将其从动脉中分离并收回。</li>
	<li>使用微钳在动脉下缝合5厘米长的无菌丝线。在动脉周围系上双结缝合线以阻塞血流。</li>
	<li>切开多余的缝合线，并通过拉回皮下组织和皮肤来关闭暴露的动脉。使用兽医粘合来密封切口。</li>
	<li>将动物放回室中在室温下的连续脑电图监测中，该室放置在加热床垫上。对幼崽核心温度进行点红外温度检查，以避免打开腔室。让动物在缺氧前恢复1小时。</li>
</ol>5. 脑电图和缺氧
<ol><li>通过氧气监测仪连续监测腔室内的FiO2 （吸入氧气的分数）。</li>
	<li>用60升/分钟的100%N2 和0.415升/分钟冲洗腔室，100%O2。一旦腔室中的氧饱和度达到12%，则将N2流量降低至10 L / min，同时保持O2 流量不变。通过小幅调整，将FiO2 保持在8%45分钟。</li>
	<li>缺氧暴露45分钟后，将FiO2 返回至21%。</li>
	<li>让幼崽在腔室中恢复，并在缺氧后监测脑电图2小时。</li>
	<li>记录期结束后，断开小鼠与脑电图记录的连接并返回给母亲。</li>
</ol>6. 脑电图分析
<ol><li>在LabChart Pro中用视频分析EEG文件。让盲法研究人员在脑电图上标记癫痫发作和背景模式17。癫痫发作被定义为持续超过 10 秒的心电事件，具有高频有节奏的锐波放电（≥3 倍基线），且有明显的演变17。</li>
	<li>让第二个盲法研究人员随机审查标记的事件以达成一致。</li>
	<li>回顾每个标记的心电图事件的相关视频，并根据新生儿啮齿动物行为癫痫发作评分进行分析16。简而言之，该评分范围为0-6（不动到严重的强直阵挛行为）。为了进一步描述癫痫发作的符号学特征，分析侧向性行为（多灶性/双侧运动 vs. 局灶性/单侧性 vs. 混合性）。</li>
	<li>创建功率频谱图。将快速傅里叶变换与大小为 1024 个数据点的余弦-贝尔数据窗口结合使用。借助 87.5% 的窗口重叠，在频谱图中创建平滑的 x 轴。将功率表示为μV218。</li>
</ol>

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我们已经提出了一种在缺氧缺血性癫痫发作期间对新生小鼠进行连续视频脑电图监测的模型。结合脑电图进行视频分析，可表征癫痫发作的符号学特征。脑电图分析允许提取功率谱图和背景振幅分析。
正确和小心地放置电极在此方案中至关重要，因为电极放置过程中的伤害或不正确的放置会显着影响结果。在损伤前评估正常的基线脑电图活动至关重要，因为在电极放置过程?...

缺氧缺血性脑病 （HIE） 是一种每年影响 1000 名新生儿中 1.5 名新生儿的疾病，是新生儿癫痫发作的最常见原因1，2。存活下来的婴儿有患各种神经残疾的风险，如脑瘫、智力残疾和癫痫3，4，5。
动物模型在理解和研究缺氧缺血和新生儿癫痫发作的病理生理学方面起着关键作用6，7。改进的Vannucci模型用于在出生后第10天诱导缺氧缺血（HI）（p10）7，8。这个年龄段的小鼠幼崽在神经学上大致翻译为完整的足月人类新生儿9。
与该损伤模型结合使用的连续视频脑电图 （EEG） 监测可进一步了解和表征新生儿缺氧缺血性癫痫发作。以前的研究已经使用各种方法来分析啮齿动物的新生儿癫痫发作，包括视频记录，有限的脑电图记录和遥测脑电图记录10，11，12，13，14，15，16。在下面的手稿中，我们深入讨论了在缺氧缺血期间记录小鼠幼崽连续视频脑电图的过程。这种用于新生儿小鼠幼崽连续视频脑电图监测的技术可以应用于各种疾病和癫痫发作模型。

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continuous video electroencephalogram during hypoxia-ischemia in neonatal mice

缺氧缺血是新生儿癫痫发作的最常见原因。动物模型对于了解新生儿癫痫发作和缺氧缺血的机制和生理学至关重要。本文介绍了一种在新生小鼠中连续进行视频脑电图（EEG）监测的方法，以检测癫痫发作并分析缺氧缺血期间的脑电图背景。同时使用视频和脑电图可以描述癫痫发作的符号学和确认癫痫发作。该方法还允许分析实验时间段内的功率谱图和EEG背景模式趋势。在这种缺氧缺血模型中，该方法允许在损伤前进行脑电图记录，以获得规范基线，并在损伤和恢复期间进行。总监测时间受到限制，因为无法将幼崽与母亲分开超过四个小时。虽然，我们在本文中使用了缺氧缺血性癫痫发作的模型，但这种用于新生儿视频脑电图监测的方法可以应用于啮齿动物的各种疾病和癫痫发作模型。

癫痫发作符号学
新生儿缺氧缺血暴露导致小鼠全身性和局灶性癫痫发作（图1A-C）。视频脑电图记录允许将电图结果与视频行为相关联。这些行为使用先前发表的新生儿啮齿动物行为癫痫发作评分（BSS）进行评分16。除BSS外，我们还根据行为是局灶性/单侧，双侧还是混合性对事件进行分类（图1B...

Watch this Scientific Journal Video about Continuous Video Electroencephalogram during Hypoxia-Ischemia in Neonatal Mice at JoVE.com

Continuous Video Electroencephalogram during Hypoxia-Ischemia in Neonatal Mice

本手稿描述了一种在经历缺氧缺血的新生小鼠中使用多个深度电极进行连续视频脑电图记录的方法。

新生小鼠缺氧缺血期间的连续视频脑电图

Hypoxia ischemia is the most common cause of neonatal seizures. Animal models are crucial for understanding the mechanisms and physiology underlying ...

continuous-video-electroencephalogram-during-hypoxia-ischemia

Universidad de Cartagena UDEC

Research

JoVE Journal

Neuroscience

3.3K 次观看.  University of Virginia. 本手稿描述了一种在经历缺氧缺血的新生小鼠中使用多个深度电极进行连续视频脑电图记录的方法。

视频: Continuous Video Electroencephalogram during Hypoxia-Ischemia in Neonatal Mice

Delivery of In Vivo Acute Intermittent Hypoxia in Neonatal Rodents to Prime Subventricular Zone-derived Neural Progenitor Cell Cultures

Extended culture of neural stem/progenitor cells facilitates in vitro analyses to understand their biology while enabling expansion of cell populations to adequate numbers prior to transplantation. Identifying approaches to refine this process, to augment the production of all CNS cell types (i.e., neurons), and to possibly contribute to therapeutic cell therapy protocols is a high research priority. This report describes an easily applied in vivo &#8220;pre-conditioning&#8221; stimulus which can be delivered to awake, non-anesthetized animals. Thus, it is a non-invasive and non-stressful procedure. Specifically described are the procedures for exposing mouse or rat pups (aged postnatal day 1-8) to a brief (40-80 min) period of intermittent hypoxia (AIH). The procedures included in this video protocol include calibration of the whole-body plethysmography chamber in which pups are placed during AIH and the technical details of AIH exposure. The efficacy of this approach to elicit tissue-level changes in the awake animal is demonstrated through the enhancement of subsequent in vitro expansion and neuronal differentiation in cells harvested from the subventricular zone (SVZ). These results support the notion that tissue level changes across multiple systems could be observed following AIH, and support the continued optimization and establishment of AIH as a priming or conditioning modality for therapeutic cell populations.

Delivery of In Vivo Acute Intermittent Hypoxia in Neonatal Rodents to Prime Subventricular Zone-derived Neural Progenitor Cell Cultures

This article describes the methodology for administering short periods of intermittent hypoxia to postnatal day 1-8 mouse or rat pups. This approach effectively elicits a robust tissue level &#8220;priming effect&#8221; on cultured neural progenitor cells that are harvested within 30 min of hypoxia exposure.

交付<em&gt;在体内</em&gt;急性间歇性缺氧新生儿鼠害总理下区源性神经祖细胞培养

Extended culture of neural stem/progenitor cells facilitates in vitro analyses to understand their biology while enabling expansion of cell populations to ...

科研

发育生物学

Simultaneous Video-EEG-ECG Monitoring to Identify Neurocardiac Dysfunction in Mouse Models of Epilepsy

In epilepsy, seizures can evoke cardiac rhythm disturbances such as heart rate changes, conduction blocks, asystoles, and arrhythmias, which can potentially increase risk of sudden unexpected death in epilepsy (SUDEP). Electroencephalography (EEG) and electrocardiography (ECG) are widely used clinical diagnostic tools to monitor for abnormal brain and cardiac rhythms in patients. Here, a technique to simultaneously record video, EEG, and ECG in mice to measure behavior, brain, and cardiac activities, respectively, is described. The technique described herein utilizes a tethered (i.e., wired) recording configuration in which the implanted electrode on the head of the mouse is hard-wired to the recording equipment. Compared to wireless telemetry recording systems, the tethered arrangement possesses several technical advantages such as a greater possible number of channels for recording EEG or other biopotentials; lower electrode costs; and greater frequency bandwidth (i.e., sampling rate) of recordings. The basics of this technique can also be easily modified to accommodate recording other biosignals, such as electromyography (EMG) or plethysmography for assessment of muscle and respiratory activity, respectively. In addition to describing how to perform the EEG-ECG recordings, we also detail methods to quantify the resulting data for seizures, EEG spectral power, cardiac function, and heart rate variability, which we demonstrate in an example experiment using a mouse with epilepsy due to Kcna1 gene deletion. Video-EEG-ECG monitoring in mouse models of epilepsy or other neurological disease provides a powerful tool to identify dysfunction at the level of the brain, heart, or brain-heart interactions.

Here, we present a protocol to record brain and heart bio signals in mice using simultaneous video, electroencephalography (EEG), and electrocardiography (ECG). We also describe methods to analyze the resulting EEG-ECG recordings for seizures, EEG spectral power, cardiac function, and heart rate variability.

同步视频脑电图-心电图监测对小鼠癫痫模型 Neurocardiac 功能障碍的鉴别

In epilepsy, seizures can evoke cardiac rhythm disturbances such as heart rate changes, conduction blocks, asystoles, and arrhythmias, which can ...

遗传学

The Hypoxic Ischemic Encephalopathy Model of Perinatal Ischemia

Hypoxic-Ischemic Encephalopathy (HIE) is the consequence of systemic asphyxia occurring at birth. Twenty five percent of neonates with HIE develop severe and permanent neuropsychological sequelae, including mental retardation, cerebral palsy, and epilepsy. The outcomes of HIE are devastating and permanent, making it critical to identify and develop therapeutic strategies to reduce brain injury in newborns with HIE. To that end, the neonatal rat model for hypoxic-ischemic brain injury has been developed to model this human condition. The HIE model was first validated by Vannucci et al 1 and has since been extensively used to identify mechanisms of brain injury resulting from perinatal hypoxia-ischemia 2 and to test potential therapeutic interventions 3,4. The HIE model is a two step process and involves the ligation of the left common carotid artery followed by exposure to a hypoxic environment. Cerebral blood flow (CBF) in the hemisphere ipsilateral to the ligated carotid artery does not decrease because of the collateral blood flow via the circle of Willis; however with lower oxygen tension, the CBF in the ipsilateral hemisphere decreases significantly and results in unilateral ischemic injury. The use of 2,3,5-triphenyltetrazolium chloride (TTC) to stain and identify ischemic brain tissue was originally developed for adult models of rodent cerebral ischemia 5, and is used to evaluate the extent of cerebral infarctin at early time points up to 72 hours after the ischemic event 6. In this video, we demonstrate the hypoxic-ischemic injury model in postnatal rat brain and the evaluation of the infarct size using TTC staining.

The postnatal rat model for hypoxic-ischemic brain injury is a well-established model of human neonatal hypoxic ischemic encephalopathy (HIE). In this article, we describe the model of HIE in post-natal rat pups.

围产期缺血缺氧缺血性脑病模型

Hypoxic-Ischemic Encephalopathy (HIE) is the consequence of systemic asphyxia occurring at birth. Twenty five percent of neonates with HIE develop severe ...

生物学

Fetal Mouse Cardiovascular Imaging Using a High-frequency Ultrasound (30/45MHZ) System

Congenital heart defects (CHDs) are the most common cause of childhood morbidity and early mortality. Prenatal detection&#160;of the underlying molecular mechanisms of CHDs is&#160;crucial for inventing new preventive and therapeutic strategies. Mutant mouse models are powerful tools to discover new&#160;mechanisms&#160;and environmental stress modifiers that drive cardiac development and their potential&#160;alteration in CHDs. However, efforts to establish the causality of these putative contributors have been limited to histological&#160;and molecular studies&#160;in non-survival animal experiments, in which&#160;monitoring the key physiological and hemodynamic parameters is&#160;often absent. Live imaging technology has become an essential tool to establish the&#160;etiology of CHDs. In particular,&#160;ultrasound imaging can be used prenatally without surgically exposing the fetuses,&#160;allowing maintaining&#160;their baseline physiology while monitoring the impact of environmental stress&#160;on the hemodynamic and structural aspects of cardiac chamber development. Herein, we use the High-Frequency Ultrasound (30/45)&#160;system to examine the cardiovascular system in fetal mice at E18.5 in utero at the baseline and in response to prenatal hypoxia exposure. We demonstrate the feasibility of the system to measure cardiac chamber size, morphology, ventricular function, fetal heart rate, and umbilical artery flow indices, and their alterations in fetal mice exposed to systemic chronic hypoxia in utero in real time.

Fetal Mouse Cardiovascular Imaging Using a High-frequency Ultrasound 30/45MHZ System

High-frequency ultrasound imaging of the fetal mouse has improved imaging resolution and can provide precise non-invasive characterization of cardiac development and structural defects. The protocol outlined herein is designed to perform real-time fetal mice echocardiography in vivo.

用高频超声 (30/45 mhz) 系统对胎儿小鼠进行心血管成像

Congenital heart defects (CHDs) are the most common cause of childhood morbidity and early mortality. Prenatal detection&#160;of the underlying molecular ...

A Piglet Model of Neonatal Hypoxic-Ischemic Encephalopathy

Birth asphyxia, which causes hypoxic-ischemic encephalopathy (HIE), accounts for 0.66 million deaths worldwide each year, about a quarter of the world&#8217;s 2.9 million neonatal deaths. Animal models of HIE have contributed to the understanding of the pathophysiology in HIE, and have highlighted the dynamic process that occur in brain injury due to perinatal asphyxia. Thus, animal studies have suggested a time-window for post-insult treatment strategies. Hypothermia has been tested as a treatment for HIE in pdiglet models and subsequently proven effective in clinical trials. Variations of the model have been applied in the study of adjunctive neuroprotective methods and piglet studies of xenon and melatonin have led to clinical phase I and II trials1,2. The piglet HIE model is further used for neonatal resuscitation- and hemodynamic studies as well as in investigations of cerebral hypoxia on a cellular level. However, it is a technically challenging model and variations in the protocol may result in either too mild or too severe brain injury. In this article, we demonstrate the technical procedures necessary for establishing a stable piglet model of neonatal HIE. First, the newborn piglet (&#60; 24 hr old, median weight 1500 g) is anesthetized, intubated, and monitored in a setup comparable to that found in a neonatal intensive care unit. Global hypoxia-ischemia is induced by lowering the inspiratory oxygen fraction to achieve global hypoxia, ischemia through hypotension and a flat trace amplitude integrated EEG (aEEG) indicative of cerebral hypoxia. Survival is promoted by adjusting oxygenation according to the aEEG response and blood pressure. Brain injury is quantified by histopathology and magnetic resonance imaging after 72 hr.

Hypoxic-ischemic encephalopathy following perinatal asphyxia can be studied using animal models. We demonstrate the procedures necessary for establishing a piglet model of neonatal hypoxic-ischemic encephalopathy.

新生儿缺氧缺血性脑病的小猪模型

Birth asphyxia, which causes hypoxic-ischemic encephalopathy (HIE), accounts for 0.66 million deaths worldwide each year, about a quarter of the ...

医学

Neurobehavioral Assessments in a Mouse Model of Neonatal Hypoxic-ischemic Brain Injury

We performed unilateral carotid artery occlusion on CD-1 mice to create a neonatal hypoxic-ischemic (HI) model and investigated the effects of neonatal HI brain injury by studying neurobehavioral functions in these mice compared to non-operated (i.e., normal) mice. During the study, Rice-Vannucci&#39;s method was used to induce neonatal HI brain damage in postnatal day 7-10 (P7-10) mice. The HI operation was performed on the pups by unilateral carotid artery ligation and exposure to hypoxia (8% O2 and 92% N2 for 90 min). One week after the operation, the damaged brains were evaluated with the naked eye through the semi-transparent skull and were categorized into subgroups based on the absence (&#34;no cortical injury&#34; group) or presence (&#34;cortical injury&#34; group) of cortical injury, such as a lesion in the right hemisphere. On week 6, the following neurobehavioral tests were performed to evaluate the cognitive and motor functions: passive avoidance task (PAT), ladder walking test, and grip strength test. These behavioral tests are helpful in determining the effects of neonatal HI brain injury and are used in other mouse models of neurodegenerative diseases. In this study, neonatal HI brain injury mice showed motor deficits that corresponded to right hemisphere damage. The behavioral test results are relevant to the deficits observed in human neonatal HI patients, such as cerebral palsy or neonatal stroke patients. In this study, a mouse model of neonatal HI brain injury was established and showed different degrees of motor deficits and cognitive impairment compared to non-operated mice. This work provides basic information on the HI mouse model. MRI images demonstrate the different phenotypes, separated according to the severity of brain damage by motor and cognitive tests.

We performed unilateral carotid artery occlusion on postnatal day 7-10 CD-1 mouse pups to create a neonatal hypoxic-ischemic (HI) model and investigated the effects of HI brain injury. We studied neurobehavioral functions in these mice compared to non-operated normal mice.

小鼠新生儿缺氧缺血性脑损伤模型的神经行为评估

We performed unilateral carotid artery occlusion on CD-1 mice to create a neonatal hypoxic-ischemic (HI) model and investigated the effects of neonatal HI ...

神经科学

Noninvasive Electrocardiography in the Perinatal Mouse

Electrocardiography (ECG) has long been relied upon as an effective and reliable method of assessing cardiovascular (and cardiopulmonary) function in both human and animal models of disease. Individual heart rate, rhythm, and regularity, combined with quantitative parameters collected from ECG, serve to assess the integrity of the cardiac conduction system as well as the integrated physiology of the cardiac cycle. This article provides a comprehensive description of the methods and techniques used to perform a noninvasive ECG on perinatal and neonatal mouse pups as early as the first postnatal day, without requiring the use of anesthetics. This protocol was designed to directly address a need for a standardized and repeatable method for obtaining ECG in newborn mice. From a translational perspective, this protocol proves to be entirely effective for characterization of congenital cardiopulmonary defects generated using transgenic mouse lines, and particularly for analysis of defects causing lethality at or during the first postnatal days. This protocol also aims to directly address a gap in the scientific literature to characterize and provide normative data associated with maturation of the early postnatal cardiac conduction system. This method is not limited to a specific postnatal timepoint, but rather allows for ECG data collection in neonatal mouse pups from birth to postnatal day 10 (P10), a window that is of critical importance for modeling human diseases in vivo, with particular emphasis on congenital heart disease (CHD).

Here, we present a noninvasive electrocardiography (ECG) protocol, optimized for early postnatal mice, that does not require the use of anesthetics.

围产期小鼠中的非侵入性心电图

Electrocardiography (ECG) has long been relied upon as an effective and reliable method of assessing cardiovascular (and cardiopulmonary) function in both ...

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

Echocardiography is a non-invasive procedure that enables the evaluation of structural and functional parameters in animal models of cardiovascular disease and is used to assess the impact of potential treatments in preclinical studies. Echocardiographic studies are usually conducted in young adult mice (i.e., 4-6 weeks of age). The evaluation of early neonatal cardiovascular function is not usually performed because of the small size of the mouse pups and the associated technical difficulties. One of the most important challenges is that the short length of the pups&#39; limbs prevents them from reaching the electrodes in the echocardiography platform. Body temperature is the other challenge, as pups are very susceptible to changes in temperature. Therefore, it is important to establish a practical guide for performing echocardiographic studies in small mouse pups to help researchers detect early pathological changes and study the progression of cardiovascular disease over time. The current work describes a protocol for performing echocardiography in mouse pups at the early age of 7 days old. The echocardiographic characterization of cardiac morphology, function, and coronary flow in neonatal mice is also described.

The present protocol describes the echocardiographic assessment of left ventricular morphology, function, and coronary blood flow in 7-day old 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 ...

Early Pathological and Magnetic Resonance Detection of Cerebral Injury Using a Rat Model of Neonatal Hypoxic Ischemic Encephalopathy

Perinatal hypoxic-ischemic encephalopathy (HIE) is an acute disease that may afflict newborns, resulting in variable long- and short-term neurodevelopmental outcomes. Early diagnosis is critical to identifying infants who may benefit from intervention; however, early diagnosis relies heavily on clinical criteria. No molecular or radiological tests have shown promise in detecting early cerebral injury. Studies have shown that magnetic resonance imaging (MRI) can show changes in both blood flow/ischemia and metabolic disruption. However, they have all been used to evaluate the secondary phase of the disease (&gt;12 h) after the onset of the injury. Early diagnosis is critical to rapidly starting therapeutic hypothermia in eligible infants, which is currently recommended to be initiated within 6 h of birth. The rat model of hypoxic-ischemic injury was developed in 1981 and has been validated and used extensively to study changes in brain perfusion, cerebral injury markers, and morphology. However, it has primarily been used as a "late model", evaluating injury several days after the initial ischemic insult. The model has been known to have poor sensitivity in evaluating reliable and reproducible early cerebral changes. The objective of this study was to develop a reliable model to study early gross morphological and radiological markers of HIE using pathological staining and cerebral magnetic resonance imaging/magnetic resonance spectroscopy.

The present protocol describes a rodent model of newborn hypoxic-ischemic injury for identifying early changes in cerebral tissue by gross morphology and magnetic resonance imaging. This has benefits over existing models, which can be used to study late injury but do not allow the evaluation of reproducible early changes.

Perinatal hypoxic-ischemic encephalopathy (HIE) is an acute disease that may afflict newborns, resulting in variable long- and short-term ...

Establishing a Mouse Model of Neonatal Hypoxic-Ischemic Brain Injury

Source: Kim, M., et al. <a href="https://appjove.unicartagenaproxy.elogim.com/v/55838">Neurobehavioral Assessments in a Mouse Model of Neonatal Hypoxic-ischemic Brain Injury.</a> J. Vis. Exp. (2017).
The video demonstrates the procedure for creating a mouse model to study neonatal hypoxic-ischemic brain injury. It outlines the steps for surgically inducing ischemia and subjecting the animal to hypoxia to exacerbate brain damage. Finally, it shows how to assess the resulting brain lesions, ensuring the mouse model is prepared for further scientific investigation.