这项工作是由美国国立卫生研究院（RO1 NS059043 RO1 ES015988），国家多发性硬化症协会，罗氏对贫血的研究，费尔德斯坦医学基金会基金会，施赖纳斯和儿童医院的拨款支持的一部分。 我们想感谢他的建议，大卫博士游乐场。这项工作是支持部分资助来自美国国立卫生研究院的WD（RO1 NS059043 RO1 ES015988），国家多发性硬化症协会，和施赖纳斯儿童医院。我们声明没有竞争利益有关对这篇文章。

在P6小鼠建立新生儿脑损伤的详细程序： <ol><li>产后第6天（六）小鼠对有害刺激的反应迟钝，以深麻醉点冰间接冷却麻醉。 </li><li>中线腹切口用酒精清洁皮肤，在颈前。 </li><li>在解剖显微镜下，右侧颈总动脉分叉处是上前轻轻收回肩胛舌骨和胸锁乳突肌。 </li><li>颈动脉鞘周围筋膜被删除，从附近的迷走神经和交感神经节与一个钩子的孤立和近端内部分支。颈内动脉，然后使用烧灼提示，烧灼。 </li><li>烧灼，皮肤切口缝合，直到完全清醒，然后返回到大坝的动物是保持温暖。 </li><li>结扎后1小时，动物被放置在一个密封腔，用氮气注入至6.0％，达到O 2的水平。该动物暴露在缺氧35分钟。 </li><li>缺氧暴露后，鼠标恢复为30分钟（33℃）加热垫，然后返回到大坝。动物是创造脑损伤合并缺氧/缺血和感染/炎症，然后注入0.015毫升脂多糖（LPS 1毫克/公斤）的腹腔。 </li><li>四天post-hypoxia/ischemia，小鼠麻醉反应迟钝点冰间接冷却有害刺激，然后用生理盐水和4％多聚甲醛灌注。大脑将被删除，并cryoprotected。 </li></ol><img src="/files/ftp_upload/1951/1951fig1.jpg" alt="图1" /> 图1脑缺氧/缺血小鼠模型与H＆E染色损伤的表征。有灶性坏死，脑白质（箭头）（一）颈动脉结扎患侧较对侧脑白质（二）。在海马（C）和丘脑（四）在患侧脑白质损伤的大脑同侧坏死和联络microinfarcts观察。 代表性的成果和数据： PVL的小鼠模型与缺氧/缺血和/或炎症感染/取得暴露于缺氧及注射LPS单方面颈内动脉结扎术。我们定义为适合PVL的模式新的小鼠模型。冠状切片染色与苏木精和曙红（H＆E）在整个前脑的神经病理检查发现大脑半球同侧颈动脉结扎（ 图 1A）在中央白质灶性坏死，神经元的相对不遗余力地覆大脑皮层相比，对侧脑白质（ 图1B）。在局灶性坏死脑白质，有显着增加巨噬细胞和反应性星形胶质细胞组成的细胞结构，局灶性人类PVL的特点。在对侧中央白质，少突胶质细胞的细胞核被安排在丛生状束，这个架构是完全由同侧灶性坏死破坏。值得注意的是，我们还观察患侧脑白质损伤的大脑同侧坏死（ 图1C）在海马和丘脑（焦点microinfarcts）（ 图1D）。人力PVL的，也不会出现在完全隔离，​​从伴随灰质病变。 PVL的标志，是相对的，虽然不是完整的，不遗余力脑皮层覆盖在受伤的白质 - 一个功能，我们认为是存在于我们的小鼠模型。 <img src="/files/ftp_upload/1951/1951fig2.jpg" alt="图2" /> 图2。评分0-5规模评估脑白质损伤，脑室周围白质软化的小鼠模型中MBP或O1。 我们的小鼠模型显示PVL的特点，同时在表型和分子水平。我们已经用髓鞘碱性蛋白（MBP）或O1 -抗原的免疫细胞化学检测，以评估损伤，已经建立了一个标准的规模从0到5，得分脑白质损伤（图2），并有特点的脑白质病理（图3）。最近，一个不带偏见的体视学测定已实施最初的半定量的方法来代替。此外，测试功能结果脑白质损伤的相关性的影响，我们已完成对运动功能的行为测试。我们已经证明，小鼠表现出惊人的运动障碍，P10 - 21以下缺氧/缺血加LPS在小六。我们建立了0-4分的评分标准来定义损伤小鼠P10对侧肢体功能障碍-21。通常情况下，正常小鼠可以炸了一个30 °倾斜（4分），并爬上45 °斜面无滑动（3分）。损伤小鼠的脑白质损伤的程度较低的相关性得分。 <img src="/files/ftp_upload/1951/1951fig3.jpg" alt="图3" /> 图3。MBP或O1的染色和电子显微镜（EM）的髓磷脂结构脑白质损伤的表征。 MBP和弧菌染色显示髓磷脂损失结扎患侧脑白质相比，对侧脑白质。 EM检查显示脑白质髓鞘的轴突结扎对侧，患侧脑白质退化的组织。 我们预计，PVL的新的小鼠模型的建立，将使我们能够研究的未成熟脑PVL的样损伤的具体机制。我们将讨论使用转基因小鼠品系的发病机制研究单个分子的利益的作用和评估的神经保护作用的目标。

<ol>
	<li>Volpe, J. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Neurology of newborn. , 217-276 (2001).</li><li>Deng, W., Pleasure, J., Pleasure, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Progress+in+periventricular+leukomalacia.">Progress in periventricular leukomalacia.</a> Arch. Neurol. 65, 1291-1295 (2008).</li><li>Follett, P. L., Deng, W., Dai, W., Talos, D. M., Massillon, L. J., Rosenberg, P. A., Volpe, J. J., Jensen, F. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Glutamate+receptor-mediated+oligodendrocyte+toxicity+in+periventricular+leukomalacia:+a+protective+role+for+topiramate.">Glutamate receptor-mediated oligodendrocyte toxicity in periventricular leukomalacia: a protective role for topiramate.</a> J. Neurosci. 24, 4412-4420 (2004).</li></ol>

发育中的大脑损伤导致破坏性的神经系统的后果。引人注目的是，围产期脑损伤的模式是高度依赖年龄。足月儿，它主要影响大脑皮层与特征的神经元损失。然而，在早产儿中，有选择性，突出发展少突胶质细胞损伤影响脑的白质，无序称为脑室周围白质软化（PVL）。在早产儿脑损伤的问题的严重性是非凡的。早产和改善新生儿重症监护，导致近90％的13万世界各地的早产儿与出生体重低于1500克，即非常低出生体重婴儿出生，每年（在美国大约56000）的生存。大约10％的幸存者随后表现出脑性麻痹，约50％的认知和行为的赤字。 PVL的是最重要的脑病理基本早产儿脑瘫。没有PVL的具体治疗目前存在的部分，因为发病机制尚未很好地理解，1,2 。 虽然PVL的病因是多因素的孕妇宫内感染，围产期缺氧/缺血被认为是一个PVL的首要原因。一个PVL的缺氧/缺血模型大鼠在P7的先前已成立3。这种模式是独一无二的，它类似于在早产儿PVL的。这种模式的特点是有选择性的脑白质病理，相反大多数中风模型的特点是由灰质梗死。然而，在大鼠缺氧缺血模型的局限性不容忽视。首先，PVL的病因通常是多方面的。除了缺氧/缺血，孕产妇，胎儿感染/炎症PVL的是紧密联系在一起。因此，结合缺氧/缺血和感染/炎症可能会创造更多PVL的类似机型。最PVL的病变（24-32孕周）的最大风险与人类发育时期相关的啮齿类动物的年龄是出生后，因为少突胶质细胞分化主要是在啮齿类动物的产后事件。由于它是不可能产生产后年龄产妇胎儿感染/炎症，它是合理的，以模拟在有关产后发育阶段的感染创造了PVL的啮齿动物模型。我们力求结合起来，在缺氧/缺血模型小鼠在小六LPS注射，以产生更多的临床相关PVL的病变。二，最优的概念证明型的致病作用的分子实验方法，必须使用转基因小鼠遗传操作比鼠标在大鼠更难。因此，我们必须寻求转换鼠标大鼠PVL的实验模型。 PVL的小鼠模型，将极大地促进1）利用现有的转基因小鼠品系的疾病的生物学和发病机制的研究; 2）进行药物试验，发现具有治疗潜力的候选人的代理人; 3）使用免疫缺陷的小鼠品系的干细胞的实用测试。

<!DOCTYPE html PUBLIC "-//W3C//DTD XHTML 1.0 Transitional//EN" "http://www.w3.org/TR/xhtml1/DTD/xhtml1-transitional.dtd">
<html xmlns="http://www.w3.org/1999/xhtml" xml:lang="en" lang="en">	
		
		<div>
		<table border="1">
		<thead>
		<tr>
		<th>Material Name</th>
		<th>Type</th>
		<th>Company</th>
		<th>Catalogue Number</th>
		<th>Comment</th>
		</tr>
		</thead>
		<tbody>
		
<tr>
<td>LPS</td>
<td>&nbsp;</td>
<td>Sigma</td>
<td>L4130</td>
<td>&nbsp;</td>
<tr>
<td>Hypoxia Chamber</td>
<td>&nbsp;</td>
<td>BioSpherix</td>
<td>A-30274</td>
<td>&nbsp;</td>
<tr>
<td>Surgical hook</td>
<td>&nbsp;</td>
<td>F.S.T.</td>
<td>10064-14</td>
<td>&nbsp;</td>		</tbody>
		</table>
		</div>

mouse models of periventricular leukomalacia

我们描述了一个小鼠模型的建立脑室周围白质软化（PVL）的协议。 PVL的是早产儿脑损伤和脑瘫最常见的前因的主要形式。 PVL的特点是突出的少突胶质细胞损伤脑室周围脑白质损伤。有或没有全身性感染/炎症缺血缺氧/ PVL的主要原因。我们使用P6小鼠创建缺氧/缺血诱导或新生儿脑损伤，无全身性感染/炎症暴露于缺氧或不注射内毒素脂多糖（LPS）单侧颈动脉结扎模式。髓鞘碱性蛋白（MBP）O1和电镜观察免疫组化显示脑白质突出的髓磷脂损失，额外伤害，海马和丘脑。 PVL的小鼠模型的建立将大大方便了使用可用的转基因小鼠品系，在一个相对较高的吞吐量的方式，以确定治疗药物的候选药物试验的传导，以及干细胞移植的测试，使用免疫缺陷的小鼠品系的疾病的发病机制的研究。

Watch this Scientific Journal Video about Mouse Models of Periventricular Leukomalacia at JoVE.com

Mouse Models of Periventricular Leukomalacia

我们建立了脑室周围白质软化（PVL），早产儿脑室周围脑白质病变​​为特征的主导性脑损伤的小鼠模型。 / /没有全身性感染的缺氧缺血PVL的主要原因。单侧颈动脉结扎和缺氧暴露/无脂多糖注射创建在P6小鼠PVL的样病变。

脑室周围白质软化的小鼠模型

We describe a protocol for establishing mouse models of periventricular leukomalacia (PVL). PVL is the predominant form of brain injury in premature ...

mouse-models-of-periventricular-leukomalacia

Research

JoVE Journal

Neuroscience

29.7K 次观看.  University of California, Davis. 我们建立了脑室周围白质软化（PVL），早产儿脑室周围脑白质病变​​为特征的主导性脑损伤的小鼠模型。 / /没有全身性感染的缺氧缺血PVL的主要原因。单侧颈动脉结扎和缺氧暴露/无脂多糖注射创建在P6小鼠PVL的样病变。

视频: Mouse Models of Periventricular Leukomalacia

A Simple Composite Phenotype Scoring System for Evaluating Mouse Models of Cerebellar Ataxia

We describe a protocol for the rapid and sensitive quantification of disease severity in mouse models of cerebella ataxia. It is derived from previously published phenotype assessments in several disease models, including spinocerebellar ataxias, Huntington s disease and spinobulbar muscular atrophy. Measures include hind limb clasping, ledge test, gait and kyphosis. Each measure is recorded on a scale of 0-3, with a combined total of 0-12 for all four measures. The results effectively discriminate between affected and non-affected individuals, while also quantifying the temporal progression of neurodegenerative disease phenotypes. Measures may be analyzed individually or combined into a composite phenotype score for greater statistical power. The ideal combination of the four described measures will depend upon the disorder in question. We present an example of the protocol used to assess disease severity in a transgenic mouse model of spinocerebellar ataxia type 7 (SCA7).
Albert R. La Spada and Gwenn A. Garden contributed to this manuscript equally.

We describe a protocol for the rapid and sensitive quantification of disease severity in mouse models of cerebellar ataxia. Measures include hind limb clasping, ledge test, gait and kyphosis. This protocol effectively discriminates between affected and non-affected individuals, and detects the progression of affected individuals over time.

一个简单的综合评分系统评估小脑共济失调的小鼠模型的表型

We describe a protocol for the rapid and sensitive quantification of disease severity in mouse models of cerebella ataxia. It is derived from previously ...

科研

神经科学

Experimental Models for Study of Retinal Pigment Epithelial Physiology and Pathophysiology

We have developed a cell culture procedure that can produce large quantities of confluent monolayers of primary human fetal retinal pigment epithelium (hfRPE) cultures with morphological, physiological and genetic characteristics of native human RPE. These hfRPE cell cultures exhibit heavy pigmentation, and electron microscopy show extensive apical membrane microvilli. The junctional complexes were identified with immunofluorescence labeling of various tight junction proteins. Epithelial polarity and function of these easily reproducible primary cultures closely resemble previously studied mammalian models of native RPE, including human. These results were extended by the development of therapeutic interventions in several animal models of human eye disease. We have focused on strategies for the removal of abnormal fluid accumulation in the retina or subretinal space. The extracellular subretinal space separates the photoreceptor outer segments and the apical membrane of the RPE and is critical for maintenance of retinal attachments and a whole host of RPE/retina interactions.

We provide a reproducible method for culturing confluent monolayers of human fetal retinal pigment epithelial cells (hfRPE) cells that exhibit morphology, physiology, polarity, and protein and gene expression patterns of adult native tissue. This work has been extended to an animal model of several eye diseases.

视网膜色素上皮的生理与病理生理学研究的实验模型

We have developed a cell culture procedure that can produce large quantities of confluent monolayers of primary human fetal retinal pigment epithelium ...

Multiple-mouse Neuroanatomical Magnetic Resonance Imaging

The field of mouse phenotyping with magnetic resonance imaging (MRI) is rapidly growing, motivated by the need for improved tools for characterizing and evaluating mouse models of human disease. MRI is an excellent modality for investigating genetically altered animals. It is capable of whole brain coverage, can be used in vivo, and provides multiple contrast mechanisms for investigating different aspects of neuranatomy and physiology. The advent of high-field scanners along with the ability to scan multiple mice simultaneously allows for rapid phenotyping of novel mutations.
Effective mouse MRI studies require attention to many aspects of experiment design. In this article, we will describe general methods to acquire quality images for mouse phenotyping using a system that images mice concurrently in shielded transmit/receive radio frequency (RF) coils in a common magnet (Bock et al., 2003). We focus particularly on anatomical phenotyping, an important and accessible application that has shown a high potential for impact in many mouse models at our imaging centre. Before we can provide the detailed steps to acquire such images, there are important practical considerations for both in vivo brain imaging (Dazai et al., 2004) and ex vivo brain imaging (Spring et al., 2007) that should be noted. These are discussed below.

Magnetic resonance imaging (MRI) has become an increasingly popular tool for examining the phenotype of genetically altered mice. This article illustrates the methods necessary to achieve high-throughput phenotyping of genetically altered mice using multiple-mouse MRI.

多鼠神经解剖学磁共振成像

The field of mouse phenotyping with magnetic resonance imaging (MRI) is rapidly growing, motivated by the need for improved tools for characterizing and ...

Olfactory Assays for Mouse Models of Neurodegenerative Disease

In many neurodegenerative diseases and particularly in Parkinson&#8217;s disease, deficits in olfaction are reported to occur early in the disease process and may be a useful behavioral marker for early detection. Earlier detection in neurodegenerative disease is a major goal in the field because this is when neuroprotective therapies have the best potential to be effective. Therefore, in preclinical studies testing novel neuroprotective strategies in rodent models of neurodegenerative disease, olfactory assessment could be highly useful in determining therapeutic potential of compounds and translation to the clinic. In the present study we describe a battery of olfactory assays that are useful in measuring olfactory function in mice. The tests presented in this study were chosen because they measure olfaction abilities in mice related to food odors, social odors, and non-social odors. These tests have proven useful in characterizing novel genetic mouse models of Parkinson&#8217;s disease as well as in testing potential disease-modifying therapies.

Impairment in olfactory function is a common feature in many neurodegenerative disorders including Parkinson, Alzheimer, and Huntington diseases. In the present article, we describe a set of tests for assessing olfaction discrimination and detection in mice that can be used to measure olfactory abilities in mouse models of neurodegenerative diseases.

嗅觉测定的神经退行性疾病的小鼠模型

In many neurodegenerative diseases and particularly in Parkinson&#8217;s disease, deficits in olfaction are reported to occur early in the disease process ...

3D Modeling of the Lateral Ventricles and Histological Characterization of Periventricular Tissue in Humans and Mouse

The ventricular system carries and circulates cerebral spinal fluid (CSF) and facilitates clearance of solutes and toxins from the brain. The functional units of the ventricles are ciliated epithelial cells termed ependymal cells, which line the ventricles and through ciliary action are capable of generating laminar flow of CSF at the ventricle surface. This monolayer of ependymal cells also provides barrier and filtration functions that promote exchange between brain interstitial fluids (ISF) and circulating CSF. Biochemical changes in the brain are thereby reflected in the composition of the CSF and destruction of the ependyma can disrupt the delicate balance of CSF and ISF exchange. In humans there is a strong correlation between lateral ventricle expansion and aging. Age-associated ventriculomegaly can occur even in the absence of dementia or obstruction of CSF flow. The exact cause and progression of ventriculomegaly is often unknown; however, enlarged ventricles can show regional and, often, extensive loss of ependymal cell coverage with ventricle surface astrogliosis and associated periventricular edema replacing the functional ependymal cell monolayer. Using MRI scans together with postmortem human brain tissue, we describe how to prepare, image and compile 3D renderings of lateral ventricle volumes, calculate lateral ventricle volumes, and characterize periventricular tissue through immunohistochemical analysis of en face lateral ventricle wall tissue preparations. Corresponding analyses of mouse brain tissue are also presented supporting the use of mouse models as a means to evaluate changes to the lateral ventricles and periventricular tissue found in human aging and disease. Together, these protocols allow investigations into the cause and effect of ventriculomegaly and highlight techniques to study ventricular system health and its important barrier and filtration functions within the brain.

Using MRI scans (human), 3D imaging software, and immunohistological analysis, we document changes to the brain&#8217;s lateral ventricles. Longitudinal 3D mapping of lateral ventricle volume changes and characterization of periventricular cellular changes that occur in the human brain due to aging or disease are then modeled in mice.

脑室周围组织的侧脑室及组织学特性对人类和鼠标3D建模

The ventricular system carries and circulates cerebral spinal fluid (CSF) and facilitates clearance of solutes and toxins from the brain. The functional ...

Using Enzyme-based Biosensors to Measure Tonic and Phasic Glutamate in Alzheimer's Mouse Models

Neurotransmitter disruption is often a key component of diseases of the central nervous system (CNS), playing a role in the pathology underlying Alzheimer's disease, Parkinson's disease, depression, and anxiety. Traditionally, microdialysis has been the most common (lauded) technique to examine neurotransmitter changes that occur in these disorders. But because microdialysis has the ability to measure slow 1-20 minute changes across large areas of tissue, it has the disadvantage of invasiveness, potentially destroying intrinsic connections within the brain and a slow sampling capability. A relatively newer technique, the microelectrode array (MEA), has numerous advantages for measuring specific neurotransmitter changes within discrete brain regions as they occur, making for a spatially and temporally precise approach. In addition, using MEAs is minimally invasive, allowing for measurement of neurotransmitter alterations in vivo. In our laboratory, we have been specifically interested in changes in the neurotransmitter, glutamate, related to Alzheimer's disease pathology. As such, the method described here has been used to assess potential hippocampal disruptions in glutamate in a transgenic mouse model of Alzheimer's disease. Briefly, the method used involves coating a multi-site microelectrode with an enzyme very selective for the neurotransmitter of interest and using self-referencing sites to subtract out background noise and interferents. After plating and calibration, the MEA can be constructed with a micropipette and lowered into the brain region of interest using a stereotaxic device. Here, the method described involves anesthetizing rTg(TauP301L)4510 mice and using a stereotaxic device to precisely target sub-regions (DG, CA1, and CA3) of the hippocampus.

Here, we describe the setup, software navigation, and data analysis for a spatially and temporally precise method of measuring tonic and phasic extracellular glutamate changes in vivo using enzyme-linked microelectrode arrays (MEA).

使用基于酶的生物传感器来衡量进补和阶段性谷氨酸在阿尔茨海默氏症小鼠模型

Neurotransmitter disruption is often a key component of diseases of the central nervous system (CNS), playing a role in the pathology underlying ...

Exploring Deep Space - Uncovering the Anatomy of Periventricular Structures to Reveal the Lateral Ventricles of the Human Brain

Anatomy students are typically provided with two-dimensional (2D) sections and images when studying cerebral ventricular anatomy and students find this challenging. Because the ventricles are negative spaces located deep within the brain, the only way to understand their anatomy is by appreciating their boundaries formed by related structures. Looking at a 2D representation of these spaces, in any of the cardinal planes, will not enable visualisation of all of the structures that form the boundaries of the ventricles. Thus, using 2D sections alone requires students to compute their own mental image of the 3D ventricular spaces. The aim of this study was to develop a reproducible method for dissecting the human brain to create an educational resource to enhance student understanding of the intricate relationships between the ventricles and periventricular structures. To achieve this, we created a video resource that features a step-by-step guide using a fiber dissection method to reveal the lateral and third ventricles together with the closely related limbic system and basal ganglia structures. One of the advantages of this method is that it enables delineation of the white matter tracts that are difficult to distinguish using other dissection techniques. This video is accompanied by a written protocol that provides a systematic description of the process to aid in the reproduction of the brain dissection. This package offers a valuable anatomy teaching resource for educators and students alike. By following these instructions educators can create teaching resources and students can be guided to produce their own brain dissection as a hands-on practical activity. We recommend that this video guide be incorporated into neuroanatomy teaching to enhance student understanding of the morphology and clinical relevance of the ventricles.

This paper demonstrates the effective use of a fiber dissection method to reveal the superficial white matter tracts and periventricular structures of the human brain, in three-dimensional space, to aid student comprehension of ventricular morphology.

探索深空-揭开人脑侧脑室的脑室结构解剖

Anatomy students are typically provided with two-dimensional (2D) sections and images when studying cerebral ventricular anatomy and students find this ...

Establishing Mouse Models for Zika Virus-induced Neurological Disorders Using Intracerebral Injection Strategies: Embryonic, Neonatal, and Adult

The Zika virus (ZIKV) is a flavivirus currently endemic in North, Central, and South America. It is now established that the ZIKV can cause microcephaly and additional brain abnormalities. However, the mechanism underlying the pathogenesis of ZIKV in the developing brain remains unclear. Intracerebral surgical methods are frequently used in neuroscience research to address questions about both normal and abnormal brain development and brain function. This protocol utilizes classical surgical techniques and describes methods that allow one to model ZIKV-associated human neurological disease in the mouse nervous system. While direct brain inoculation does not model the normal mode of virus transmission, the method allows investigators to ask targeted questions concerning the consequence after ZIKV infection of the developing brain. This protocol describes embryonic, neonatal, and adult stages of intraventricular inoculation of ZIKV. Once mastered, this method can become a straightforward and reproducible technique that only takes a few hours to perform.

Here we describe a method for establishing a model of Zika virus-induced microcephaly in mouse. This protocol includes methods for embryonic, neonatal, and adult-stage intracerebral inoculation of the Zika virus.

应用脑注射策略建立小鼠 Zika 病毒诱发神经系统疾病模型: 胚胎、新生儿和成人

The Zika virus (ZIKV) is a flavivirus currently endemic in North, Central, and South America. It is now established that the ZIKV can cause microcephaly ...

Adult Mouse DRG Explant and Dissociated Cell Models to Investigate Neuroplasticity and Responses to Environmental Insults Including Viral Infection

This protocol describes an ex vivo model of mouse-derived dorsal root ganglia (DRG) explant and in vitro DRG-derived co-culture of dissociated sensory neurons and glial satellite cells. These are useful and versatile models to investigate a variety of biological responses associated with physiological and pathological conditions of the peripheral nervous system (PNS) ranging from neuron-glial interaction, neuroplasticity, neuroinflammation, and viral infection. The usage of DRG explant is scientifically advantageous compared to simplistic single cells models for multiple reasons. For instance, as an organotypic culture, the DRG explant allows ex vivo transfer of an entire neuronal network including the extracellular microenvironment that play a significant role in all the neuronal and glial functions. Further, DRG explants can also be maintained ex vivo for several days and the culture conditions can be perturbed as desired. In addition, the harvested DRG can be further dissociated into an in vitro co-culture of primary sensory neurons and satellite glial cells to investigate neuronal-glial interaction, neuritogenesis, axonal cone interaction with the extracellular microenvironment, and more general, any aspect associated with the neuronal metabolism. Therefore, the DRG-explant system offers a great deal of flexibility to study a wide array of events related to biological, physiological, and pathological conditions in a cost-effective manner.

In this report, the advantages of organotypic cultures and dissociated primary cultures of mouse-derived dorsal root ganglia are highlighted to investigate a wide range of mechanisms associated with neuron-glial interaction, neuroplasticity, neuroinflammation, and response to viral infection.

成年小鼠背根节外植体和分离细胞模型调查可塑性和对包括病毒感染在内的环境侮辱的反应

This protocol describes an ex vivo model of mouse-derived dorsal root ganglia (DRG) explant and in vitro DRG-derived co-culture of dissociated sensory ...

In Vivo Electrophysiological Measurement of Compound Muscle Action Potential from the Forelimbs in Mouse Models of Motor Neuron Degeneration

Assessing the functionality of the nerve axon provides detailed information on the progression of neuromuscular disorders. Electrophysiological recordings provide a sensitive approach to measure nerve conduction in humans and rodent models. To broaden the technical possibilities for electromyography in mice, the measurement of compound muscle action potentials (CMAPs) from the brachial plexus nerve in the forelimb using needle electrodes is described here. CMAP recordings after stimulating the sciatic nerve in hindlimbs have been previously described. The newly introduced method here allows for the evaluation of the nerve conductivity at an additional site, and thus provides a more profound overview of the neuromuscular functionality. The technique provides information on both the relative number of functional axons and the myelination level. Thereby, this method can be applied to assess both axonal diseases as well as demyelinating conditions. This minimally invasive method does not require extraction of the nerve and therefore it is suitable for repeated measurements for longitudinal follow-up in the same animal. Similar recordings are performed in clinical setups to emphasize the translational relevance of the method.

In Vivo Electrophysiological Measurement of Compound Muscle Action Potential from the Forelimbs in Mouse Models of Motor Neuron Degeneration

The measurement of nerve conduction is a useful tool to assess mouse models of neurodegeneration but it is frequently only applied to stimulate the sciatic nerve in hindlimbs. Here, we describe a technique to measure compound muscle action potential (CMAP) in vivo in the mouse forelimb muscles innervated by the brachial plexus.

体内运动神经元变性小鼠前肢肌作用电位的电生理测量

Assessing the functionality of the nerve axon provides detailed information on the progression of neuromuscular disorders. Electrophysiological recordings ...