Name: mouse models of periventricular leukomalacia
Uploaded: 2010-05-18T17:00:00
Duration: 6 min 24 s
Description: Watch this Scientific Journal Video about Mouse Models of Periventricular Leukomalacia at JoVE.com

この作品は、国立衛生研究所（RO1 NS059043とRO1 ES015988）、全国多発性硬化症協会、貧血の研究、フェルドスタイン医学財団のためのロシュ財団、とシュライナーズ子ども病院からの補助金によってサポートされている部分になります。 我々は彼の提案のためにデビッドの喜びを感謝したいと思います。この作品は、部分的に国立衛生研究所からWDへの補助金（RO1 NS059043とRO1 ES015988）、国立多発性硬化症協会、とシュライナーズ子ども病院によってサポートされていました。我々はこの記事に関連する一切の競合する利害を宣言しません。

P6マウスの新生児の脳損傷の作成の詳細な手順： <ol><li>生後6日目（P6）のマウスは、有害な刺激への無反応のポイント（深麻酔）に氷の上で間接冷却で麻酔する。 </li><li>アルコールで皮膚を洗浄に続いて、正中腹側切開は、前頸部で作られています。 </li><li>解剖顕微鏡下で、右総頸動脈の分岐部を静かに後退肩甲舌骨と胸鎖乳突筋の筋肉をが近づいています。 </li><li>頸動脈鞘の周囲筋膜が除去され、近位内部の分岐は、フックに近い迷走神経および交感神経節から分離されています。内頸動脈は、焼灼先端を使用して焼灼しています。 </li><li>焼灼に続いて、皮膚の切開を縫合され、そして動物は完全に目覚めまで保温し、ダムに戻されます。 </li><li>一時間結紮した後、動物は、O 2に到達すると6.0％のレベルまで窒素を流し込んで密封されたチャンバ内に配置されます。動物は、その後、低酸素症の35分に公開されます。 </li><li>低酸素暴露後、マウスは30分間加熱パッド（33℃）に回復し、ダムに戻されます。を組み合わせた低酸素/虚血や感染/炎症と脳損傷の作成のために、動物は、リポ多糖の0.015ミリリットル（LPS、1 mg / kg）の腹腔内に注入されます。 </li><li> post-hypoxia/ischemia 4日間、マウスはその後、その後、生理食塩水と4％パラホルムアルデヒドで灌流侵害刺激への無応答のポイントまで氷上で間接冷却で麻酔する。脳を除去し、凍結保護されています。 </li></ol><img src="/files/ftp_upload/1951/1951fig1.jpg" alt="図1" /> 図1 H＆E染色による虚血/低酸素マウスモデルにおける脳損傷の特性評価。同側の大脳白質に巣状壊死（矢印）（）対側大脳白質（B）に比べて、頸動脈結紮することがあります。同側の壊死と焦点microinfarctsは海馬（C）と同側の大脳白質損傷の脳の視床（D）で観察されています。 代表的な結果と図： 低酸素/虚血および/または感染/炎症とPVLのマウスモデルは、低酸素とLPSの注射への曝露が続く片側頸動脈結紮することにより達成されています。我々は適切なPVLモデルとして新しいマウスモデルを定義している。全体の前脳を通してヘマトキシリンとエオジン（H＆E）で染色された冠状切片の神経病理学的検査では、ニューロンの相対的な温存と、頸動脈結紮（ 図1A）に同側の大脳半球の中央の白質に巣状壊死が明らかに覆う大脳皮質は、対側大脳白質（ 図1B）に比べて。焦点人間のPVLの特徴であるとして焦点壊死性白質で、マクロファージと反応性星状細胞で構成される細胞数の顕著な増加は、そこです。反対側の中央の白質では、オリゴデンドログリアの核は束のような束に配置され、このアーキテクチャは、完全に同側で巣状壊死が中断されます。注目すべきは、我々はまた、同側の大脳白質損傷のある脳で同側の海馬の壊死（ 図1C）と視床（フォーカルmicroinfarcts）（ 図1D）を観察します。人間のPVLはまた灰白質の病変を伴うから、完全な分離では発生しません。我々はマウスモデルに存在すると信じている機能 - PVLの特徴は、負傷した白質を覆う大脳皮質の温存、完全ではないが、相対的なものです。 <img src="/files/ftp_upload/1951/1951fig2.jpg" alt="図2" /> 図2脳室周囲白質軟化症のマウスモデルでMBPまたはO1による白質傷害を評価するための0-5のスコアリングスケール。 私たちのマウスモデルは、表現型と分子レベルの両方でPVLの特性を示します。我々は、傷害を評価するためにミエリン塩基性タンパク質（MBP）またはO1 -抗原の免疫細胞化学的検出を使用している白質の損傷を（ 図2）スコアを0から5の標準的な規模を確立している、と白質病変（ 図を特徴づけている3）。最近では、非バイアスされた立体的アッセイは、初期の半定量的方法の代わりに実装されています。さらに、機能的転帰の白質損傷の相関の効果をテストするために、我々は、運動機能のための行動テストを実施している。我々は、マウスがP6でP10 - 21以下の低酸素/虚血に加えてLPSで印象的な運動障害を示すことを明らかにした。我々はP10で負傷したマウスでは対側肢の機能障害を定義するために0〜4の尺度で得点基準を確立している-21。通常は、正常なマウスは°傾斜（4のスコアを）30まで走ると滑りなしの45 °傾斜（3のスコアを）登ることができます。負傷したマウスは、白質傷害の程度と相関の低いスコアを持っている。 <img src="/files/ftp_upload/1951/1951fig3.jpg" alt="図3" /> 図3。MBPまたは電子顕微鏡（EM）によるO1染色し、ミエリン構造による白質傷害のキャラクタリゼーション。 MBPと反対側の白質と比較してライゲーションに同側の大脳白質におけるO1の染色ショーミエリンの喪失、。 EMは、検査は、ライゲーションに対側大脳白質の有髄軸索を示し、同側の白質の組織を変性。 我々は、PVLの新しいマウスモデルの確立は、私たちは未熟な脳にPVLのような傷害の特定のメカニズムを研究できるようになると期待しています。我々は興味の個々の分子の役割を検討し、神経保護の目標を評価するトランスジェニックマウス系統を用いて病因に対処します。

<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>

利害の衝突は宣言されません。

発展途上の脳への損傷は、壊滅的な神経学的な結果につながります。驚くべきことに、周産期の脳損傷のパターンは非常に年齢依存性である。正期産児では、それは主に特徴的な神経細胞の損失と大脳皮質に影響を与えます。しかし、未熟児で、それが選択的に開発するオリゴデンドロサイトへの顕著な損傷と大脳白質に影響を与える、障害は脳室周囲白質軟化症（PVL）と呼ばれる。未熟児の脳損傷の問題の大きさは尋常ではない。早産と改善された新生児集中治療室は、1500グラムの下で出生時体重で毎年生まれた世界的な1300万未熟児（約56000米国では）、すなわち、非常に低出生体重児の生存率の90％近くにつながっている。生存者の約10％はその後、脳性麻痺を示し、約50％は認知行動障害を持っている。 PVLは、未熟児で最も重要な脳の病理学、基礎となる脳性麻痺です。病因はよく1,2を理解されていないため、PVLのための特定の治療法は、現在一部で、存在しない。 PVLの病因は多因子的であるが、母親の子宮内感染と周産期低酸素/虚血は、PVLの主な原因と考えられている。 P7でのラットにおけるPVLの低酸素/虚血モデルは、以前は次の3確立されている。このモデルは、未熟児におけるPVL似ている点でユニークです。このモデルの特徴は、灰白質の梗塞によって特徴付けられるストロークモデルの大部分とは対照的に、選択的な白質病変です。しかし、ラットにおける低酸素性虚血モデルの限界を見落としてはならない。最初に、PVLの病因はしばしば多因子です。低酸素/虚血、母体胎児感染/炎症に加えて強くPVLに関連付けられています。従って、複合低酸素/虚血や感染/炎症は、おそらくより多くのPVL -のようなモデルを作成します。オリゴデンドログリアの分化は、主にげっ歯類における出生後のイベントなのでベストはPVL病変（妊娠の24〜32週）の最大のリスクの人間の発育期間と相関する齧歯類の年齢は生後です。それは出生後年齢の母体胎児感染/炎症を生成することは不可能であるとして、それはPVLの齧歯類モデルを作成するために、関連する生後発達の段階で感染をシミュレートするために合理的であろう。我々は、より臨床的に関連PVL病変を生成するために、P6でのマウスの低酸素/虚血モデルとLPSの注射を組み合わせることが求​​められている。第二に、分子の原因となる役割のための最適の概念実証実験的なアプローチは、トランスジェニックマウスの使用を必要とする、遺伝子操作はマウスよりもラットではるかに困難です。従って、我々は、ラットからマウスにPVLの我々の実験モデルを変換しようとしてきた。 PVLのマウスモデルは、大きく1）疾患の生物学および病因の研究が可能なトランスジェニックマウス系統を用いて容易にする、2）薬剤の臨床試験の伝導を治療法としての可能性に候補薬剤を発見するために、免疫不全マウス系統を用いて幹細胞の有用性3）テスト。

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mouse models of periventricular leukomalacia

我々は、脳室周囲白質軟化症（PVL）のマウスモデルを確立するためのプロトコルについて説明します。 PVLは早産児と脳性麻痺の最も一般的な先行詞の脳損傷の主要な形態である。 PVLは、著名なオリゴデンドログリア損傷と脳室周囲白質の損傷によって特徴づけられる。全身感染症/炎症の有無にかかわらず、低酸素/虚血は、PVLの主な原因です。我々は、エンドトキシンのリポ多糖の注入の有無にかかわらず、低酸素への曝露（LPS）に続く片側頸動脈結紮による全身感染症/炎症の有無にかかわらず、低酸素/虚血の誘導により、新生児の脳損傷のモデルを作成するためにP6マウスを使用してください。ミエリン塩基性タンパク質（MBP）またはO1および電子顕微鏡検査の免疫組織化学では、海馬と視床への追加ダメージと大脳白質の顕著なミエリンの喪失を示しています。 PVLのマウスモデルの確立は、大幅に利用可能なトランスジェニックマウス系統を用いて疾患の病因の研究、候補治療薬を、および免疫不全マウス系統を用いた幹細胞移植のテストを識別するために、相対的に高いスループットの方法で薬剤の臨床試験の伝導を容易にするでしょう。

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

Mouse Models of Periventricular Leukomalacia

我々は、脳室周囲白質軟化症（PVL）、脳室周囲白質病変を特徴と未熟児で優勢な脳損傷のマウスモデルを確立した。全身感染症有/無低酸素/虚血は、PVLの主な原因です。リポ多糖注入なし/付き片側頸動脈結紮及び低酸素曝露がP6マウスのPVL様病変を作成します。

脳室周囲白質軟化症のマウスモデル

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

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.

小脳性運動失調のマウスモデルを評価するための単純な複合表現型のスコアリングシステム

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

神経科学

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.

網膜色素上皮生理学と病態の研究のための実験的モデル

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.

複数のマウスの神経解剖学的磁気共鳴イメージング

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.

神経変性疾患のマウスモデルのための嗅覚アッセイ

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.

ヒトとマウスにおける脳室周囲の組織の側脳室および組織学的特性の3次元モデリング

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.

Using Enzyme-based Biosensors to Measure Tonic and Phasic Glutamate in Alzheimer&#039;s Mouse Models

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).

アルツハイマー病のマウスモデルにおけるトニックと相性グルタミン酸を測定するための酵素ベースのバイオセンサーを使用して

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.

人間の脳の側脳室を明らかにする脳室周囲構造の解剖学を暴く - 深宇宙の探索

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.

脳内注入戦略を使用してジカ ウイルス性神経疾患のマウスモデルの確立: 胎生期、新生児、およびアダルト

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.

アダルト マウス後根神経節外植体し、解離細胞モデル神経可塑性とウイルス感染を含む環境の侮辱への応答を調査するため

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.

生体内で運動ニューロンの変性症のマウスモデルの前肢からの複合筋活動電位の電気生理学的測定

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