Name: ビデオ: Video-oculography in Mice
Uploaded: 2012-07-19T12:00:00
Duration: 9 min 43 s

我々は、親切に健康研究と開発のためのオランダ機構（MDJ、CDZ）、科学研究のためのオランダ機構（CDZ）、NeuroBasic（CDZ）に感謝、ベアトリクスフォン（CDZ）、SENSOPAC（CDZ）、C7（CDZ）とはPrinses彼らの財政支援のための欧州共同体のCEREBNET（CDZ）プログラムです。

<ol>
	<li>Collewijn, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Optokinetic+and+vestibulo-ocular+reflexes+in+dark-reared+rabbits.">Optokinetic and vestibulo-ocular reflexes in dark-reared rabbits.</a> Exp. Brain Res. 27, 287 (1977).</li><li>Collewijn, H. E. y. e. -. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=and+head+movements+in+freely+moving+rabbits.">and head movements in freely moving rabbits.</a> J. Physiol. 266, 471 (1977).</li><li>Collewijn, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> The oculomotor system of the rabbit and its plasticity. , (1981).</li><li>Fuller, J. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Linkage+of+eye+and+head+movements+in+the+alert+rabbit.">Linkage of eye and head movements in the alert rabbit.</a> Brain Res. 194, 219 (1980).</li><li>Buttner-Ennever, J. A., Horn, A. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Anatomical+substrates+of+oculomotor+control.">Anatomical substrates of oculomotor control.</a> Curr. Opin. Neurobiol. 7, 872 (1997).</li><li>Robinson, D. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+use+of+control+systems+analysis+in+the+neurophysiology+of+eye+movements.">The use of control systems analysis in the neurophysiology of eye movements.</a> Annu. Rev. Neurosci. 4, 463 (1981).</li><li>Robinson, D. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+purpose+of+eye+movements.">The purpose of eye movements.</a> Invest. Ophthalmol. Vis. Sci. 17, 835 (1978).</li><li>Stahl, J. S., van Alphen, A. M., De Zeeuw, C. I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+comparison+of+video+and+magnetic+search+coil+recordings+of+mouse+eye+movements.">A comparison of video and magnetic search coil recordings of mouse eye movements.</a> J. Neurosci. Methods. 99, 101 (2000).</li><li>De Zeeuw, C. I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Expression+of+a+protein+kinase+C+inhibitor+in+Purkinje+cells+blocks+cerebellar+LTD+and+adaptation+of+the+vestibulo-ocular+reflex.">Expression of a protein kinase C inhibitor in Purkinje cells blocks cerebellar LTD and adaptation of the vestibulo-ocular reflex.</a> Neuron. 20, 495 (1998).</li><li>Picciotto, M. R., Wickman, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Using+knockout+and+transgenic+mice+to+study+neurophysiology+and+behavior.">Using knockout and transgenic mice to study neurophysiology and behavior.</a> Physiol. Rev. 78, 1131 (1998).</li><li>Oommen, B. S., Stahl, J. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Eye+orientation+during+static+tilts+and+its+relationship+to+spontaneous+head+pitch+in+the+laboratory+mouse.">Eye orientation during static tilts and its relationship to spontaneous head pitch in the laboratory mouse.</a> Brain. Res. 1193, 57 (2008).</li><li>Stahl, J. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Calcium+Channelopathy+Mutants+and+Their+Role+in+Ocular+Motor.">Calcium Channelopathy Mutants and Their Role in Ocular Motor.</a> Research. Ann. N.Y. Acad. Sci. 956, 64 (2002).</li><li>Stahl, J. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Eye+movements+of+the+murine+P/Q+calcium+channel+mutant+tottering,+and+the+impact+of+aging.">Eye movements of the murine P/Q calcium channel mutant tottering, and the impact of aging.</a> J. Neurophysiol. 95, 1588 (2006).</li><li>Andreescu, C. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Estradiol+improves+cerebellar+memory+formation+by+activating+estrogen+receptor+beta.">Estradiol improves cerebellar memory formation by activating estrogen receptor beta.</a> Journal of Neuroscience. 27, 10832 (2007).</li><li>Katoh, A., Kitazawa, H., Itohara, S., Nagao, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dynamic+characteristics+and+adaptability+of+mouse+vestibulo-ocular+and+optokinetic+response+eye+movements+and+the+role+of+the+flocculo-olivary+system+revealed+by+chemical+lesions.">Dynamic characteristics and adaptability of mouse vestibulo-ocular and optokinetic response eye movements and the role of the flocculo-olivary system revealed by chemical lesions.</a> Proc. Natl. Acad. Sci. U.S.A. 95, 7705 (1998).</li><li>Stahl, J. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Using+eye+movements+to+assess+brain+function+in+mice.">Using eye movements to assess brain function in mice.</a> Vision Res. 44, 3401 (2004).</li><li>Koekkoek, S. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Gain+adaptation+and+phase+dynamics+of+compensatory+eye+movements+in+mice.">Gain adaptation and phase dynamics of compensatory eye movements in mice.</a> Genes Funct. 1, 175 (1997).</li><li>Faulstich, B. M., Onori, K. A., du Lac, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Comparison+of+plasticity+and+development+of+mouse+optokinetic+and+vestibulo-ocular+reflexes+suggests+differential+gain+control+mechanisms.">Comparison of plasticity and development of mouse optokinetic and vestibulo-ocular reflexes suggests differential gain control mechanisms.</a> Vision Res. 44, 3419 (2004).</li><li>Koekkoek, S. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Gain+adaptation+and+phase+dynamics+of+compensatory+eye+movements+in+mice.">Gain adaptation and phase dynamics of compensatory eye movements in mice.</a> Genes Funct. 1, 175 (1997).</li></ol>

利害の衝突が宣言されません。

マウスでは高品質のビデオ眼球運動記録を得るためにいくつかの要件が必要です。キャリブレーションの手順は、上記の標準化された問題で実行する必要があります。たとえば、オフセンターキャリブレーションは、瞳孔がキャリブレーション手順の間に参照CR垂直正中線上に配置されていない場合、RPの過小評価と眼球運動の結果、過大評価になります。非常に安定した瞳孔の大きさを示す試験が非常にまれであるため、さらに、我々は、キャリブレーション手順12に瞳孔サイズの補正法を統合することをお勧めします。裁判中であっても、小さなストレスはすでに大幅に瞳孔径を変更することができます。 年齢13,18、性別、14と15,16系統：眼球運動の実験を設計する際には、次の要因は、それらが眼球運動の応答に影響を与えることが知られているので考慮してかに制御する必要があります、19。瞳孔と虹彩とのコントラストは、BALB / cマウスのように、低すぎると瞳孔検出と追跡は不可能であるため、さらに、実験動物では、色素沈着虹彩を持つ必要があります。非常に緊張したり不安に動物が実験的なセットアップと拘束条件に慣れるには、実験前に、訓練する必要があります。この動物は目の小さい閉鎖または半閉鎖にプロシージャの結果を処理し、実験中に目流体の発生を防止し、その結果、優れた生徒の追跡が行われます。 最後に、データを取得し、分析する動物ごとに2〜3時間を必要とします。したがって、眼球運動の記録は、おそらく選択したマウスに適用され、ハイスループットスクリーニングテストとしては適していません具体的な手順のままになります。

<table border="1"><tbody><tr><td> 試薬の名前 </td><td> 会社 </td><td> カタログ番号 </td></tr><tr><td> Isofluran </td><td>ローディアOrganiqueファイン株式会社</td><td></td></tr><tr><td>加温パッド</td><td> FHC </td><td> 40-90-8 </td></tr><tr><td> Duratears </td><td>アルコン</td><td></td></tr><tr><td>リン酸ゲル</td><td>カー</td><td> 31297 </td></tr><tr><td>オプチボンド素数</td><td>カー</td><td> 35369 </td></tr><tr><td>オプチボンド接着剤</td><td>カー</td><td> 35369 </td></tr><tr><td>カリスマ性複合材料</td><td>ヘレウスKulzer </td><td></td></tr><tr><td>マキシマ480光硬化ユニット</td><td>ヘンリーシャイン</td><td></td></tr><tr><td> ACサーボ制御モーター</td><td>ハーモニックドライブAG </td><td></td></tr><tr><td> CYLindric画面</td><td></td><td></td></tr><tr><td>ハロゲンライト（20 W） </td><td> RSコンポーネント</td><td></td></tr><tr><td>ポテンショメータ（精密） </td><td> BOURNS株式会社。 </td><td> 6574 </td></tr><tr><td>パワー1401（I / Oインタフェース） </td><td> CED限られた</td><td></td></tr><tr><td>コンピュータ</td><td>デル</td><td></td></tr><tr><td>赤外線emmitters </td><td> RSコンポーネント</td><td> 195から451 </td></tr><tr><td> ETL-200 </td><td> ISCAN </td><td></td></tr><tr><td>ズームレンズ（ズーム6000） </td><td> Navitar社株式会社。 </td><td></td></tr><tr><td> Pilocarpinenitrate（minims） </td><td>ラボラトリーショーバン</td><td></td></tr></tbody></table>

video-oculography in mice

目の動きは、オブジェクトを追跡したり、移動中の網膜上に画像を安定させるために非常に重要です。マウスなどの窩ない動物は、ターゲットに彼らの目をロックするための限られた能力を持っています。これらの目標に向け、目の動きとは対照的に、代償性眼眼球運動が容易にafoveate動物1,2,3,4で誘発されています。代償性眼球運動は目の筋肉を駆動するコマンド信号に前庭と視運動情報を処理することによって生成されます。前庭と視運動情報の処理は、眼球システムの赤字の仕様を可能にする、別々と一緒に調査することができます。動眼神経システムは、視運動性反射（OKR）、前庭動眼反射（VOR）または視覚的に強化された前庭動眼反射（VVOR）を想起させることによってテストすることができます。 VORが反射目のmであるのに対し、OKRは、網膜上で &quot;フルフィールド&quot;画像の動きを補正する反射運動である頭部の動きを補償ovement。 VVORは、適切な補償を行うだけでなく、視運動性の情報を両方の前庭を使用して反射眼球運動である。小脳では、監視し、これらの代償性眼球の動きを調整することができます。したがって、眼球運動記録法だけでな​​く、病的状態（前庭、眼および/または小脳由来のFe）の下で、通常の下で脳と行動の関係を調べるために非常に強力なツールです。 動眼神経システムをテストし、行動のパラダイムとして、いくつかの理由で興味深いものです。まず、眼球運動システムはよく理解神経系5である。第二に、眼球運動システムは相対的なシンプルな6であり;可能性眼球運動の量は、そのボール·イン·ソケット·アーキテクチャー（ &quot;シングル·ジョイント&quot;）と外眼筋の三対7で制限されています。第三に、行動出力と感覚入力を簡単に定量するためのこの非常にアクセス可能なシステムをなす、測定することができます分析8。多くの行動テストでは、定量的な力のこのような高いレベルを欠いている。そして最後に、パフォーマンスだけでなく、眼球運動系の可塑性の両方が学習と記憶のプロセス9の研究を可能にする、テストすることができます。 遺伝子改変マウスは、今日広く利用可能であり、彼らは様々なレベル10での脳機能の探査のための重要な供給源を形成します。加えて、それらはヒト疾患を模倣するモデルとして使用することができます。正常に眼球運動記録法、薬理学的に処理または遺伝子改変マウスを適用すると、正常および病的条件下でモーターの動作の基本的な生理機能を探索する強力な研究ツールです。ここでは、マウス8でビデオ眼球運動記録法を測定する方法について説明します。

Watch this Scientific Journal Video about Video-oculography in Mice at JoVE.com

Video-oculography in Mice

ビデオ眼球運動記録法は、眼球運動性能だけでなく、運動学習を調査するために非常に定量的な方法です。ここでは、マウスでビデオ眼球運動記録法を測定する方法について説明します。通常、薬理学的に処理または遺伝子改変マウスで、この手法を適用すると、モーターの動作の基本的な生理機能を探索する強力な研究ツールです。

マウスのビデオ眼球運動記録法

Eye movements are very important in order to track an object or to stabilize an image on the retina during movement. Animals without a fovea, such as the ...

Research

JoVE Journal

Neuroscience

23.5K ビュー.  Erasmus MC, Rotterdam, The Netherlands. ビデオ眼球運動記録法は、眼球運動性能だけでなく、運動学習を調査するために非常に定量的な方法です。ここでは、マウスでビデオ眼球運動記録法を測定する方法について説明します。通常、薬理学的に処理または遺伝子改変マウスで、この手法を適用すると、モーターの動作の基本的な生理機能を探索する強力な研究ツールです。

ビデオ: Video-oculography in Mice

Measuring Motor Coordination in Mice

Mice are increasingly being used in behavioral neuroscience, largely replacing rats as the behaviorist's animal of choice. Before aspects of behavior such as emotionality or cognition can be assessed, however, it is vital to determine whether the motor capabilities of e.g. a mutant or lesioned mouse allow such an assessment. Performance on a maze task requiring strength and coordination, such as the Morris water maze, might well be impaired in a mouse by motor, rather than cognitive, impairments, so it is essential to selectively dissect the latter from the former. For example, sensorimotor impairments caused by NMDA antagonists have been shown to impair water maze performance2. 
Motor coordination has traditionally been assessed in mice and rats by the rotarod test, in which the animal is placed on a horizontal rod that rotates about its long axis; the animal must walk forwards to remain upright and not fall off. Both set speed and accelerating versions of the rotarod are available.
The other three tests described in this article (horizontal bar, static rods and parallel bars) all measure coordination on static apparatus. The horizontal bar also requires strength for adequate performance, particularly of the forelimbs as the mouse initially grips the bar just with the front paws. 
Adult rats do not perform well on tests such as the static rods and parallel bars (personal observations); they appear less well coordinated than mice. I have only tested male rats, however, and male mice seem generally less well coordinated than females. Mice appear to have a higher strength:weight ratio than rats; the Latin name, Mus musculus, seems entirely appropriate. The rotarod, the variations of the foot fault test12 or the Catwalk (Noldus)15 apparatus are generally used to assess motor coordination in rats.

Protocols are presented for two established motor coordination tasks, the accelerating rotarod and horizontal bar, also two tests developed in Oxford recently, the static rods and parallel bars. These tests can detect motor impairments potentially of interest in their own right, as well as being possible variables in tests of other areas of behavior.

マウスにおける運動協調性の測定

Mice are increasingly being used in behavioral neuroscience, largely replacing rats as the behaviorist's animal of choice. Before aspects of behavior such ...

神経科学

A Simple and Inexpensive Method for Determining Cold Sensitivity and Adaptation in Mice

Cold hypersensitivity is a serious clinical problem, affecting a broad subset of patients and causing significant decreases in quality of life. The cold plantar assay allows the objective and inexpensive assessment of cold sensitivity in mice, and can quantify both analgesia and hypersensitivity. Mice are acclimated on a glass plate, and a compressed dry ice pellet is held against the glass surface underneath the hindpaw. The latency to withdrawal from the cooling glass is used as a measure of cold sensitivity.
Cold sensation is also important for survival in regions with seasonal temperature shifts, and in order to maintain sensitivity animals must be able to adjust their thermal response thresholds to match the ambient temperature. The Cold Plantar Assay (CPA) also allows the study of adaptation to changes in ambient temperature by testing the cold sensitivity of mice at temperatures ranging from 30 &#176;C to 5 &#176;C. Mice are acclimated as described above, but the glass plate is cooled to the desired starting temperature using aluminum boxes (or aluminum foil packets) filled with hot water, wet ice, or dry ice. The temperature of the plate is measured at the center using a filament T-type thermocouple probe. Once the plate has reached the desired starting temperature, the animals are tested as described above.
This assay allows testing of mice at temperatures ranging from innocuous to noxious. The CPA yields unambiguous and consistent behavioral responses in uninjured mice and can be used to quantify both hypersensitivity and analgesia. This protocol describes how to use the CPA to measure cold hypersensitivity, analgesia, and adaptation in mice.

The Cold Plantar Assay (CPA) measures cold responsiveness between 30 &#176;C and 5 &#176;C, and can also measure cold adaptation. This protocol describes how to use the CPA to measure cold hypersensitivity, analgesia, and adaptation in mice.

マウスにおける低温感受性と適応を決定するためのシンプルで安価な方法

Cold hypersensitivity is a serious clinical problem, affecting a broad subset of patients and causing significant decreases in quality of life.  The cold ...

Assessment of Dopaminergic Homeostasis in Mice by Use of High-performance Liquid Chromatography Analysis and Synaptosomal Dopamine Uptake

Dopamine (DA) is a modulatory neurotransmitter controlling motor activity, reward processes and cognitive function. Impairment of dopaminergic (DAergic) neurotransmission is strongly associated with several central nervous system-associated diseases such as Parkinson&#39;s disease, attention-deficit-hyperactivity disorder and drug addiction1,2,3,4. Delineating disease mechanisms involving DA imbalance is critically dependent on animal models to mimic aspects of the diseases, and thus protocols that assess specific parts of the DA homeostasis are important to provide novel insights and possible therapeutic targets for these diseases.
Here, we present two useful experimental protocols that when combined provide a functional read-out of the DAergic system in mice. Biochemical and functional parameters on DA homeostasis are obtained through assessment of DA levels and dopamine transporter (DAT) functionality5. When investigating the DA system, the ability to reliably measure endogenous levels of DA from adult brain is essential. Therefore, we present how to perform high-performance liquid chromatography (HPLC) on brain tissue from mice to determine levels of DA. We perform the experiment on tissue from dorsal striatum (dStr) and nucleus accumbens (NAc), but the method is also suitable for other DA-innervated brain areas.
DAT is essential for reuptake of DA into the presynaptic terminal, thereby controlling the temporal and spatial activity of released DA. Knowing the levels and functionality of DAT in the striatum is of major importance when assessing DA homeostasis. Here, we provide a protocol that allows to simultaneously deduce information on surface levels and function using a synaptosomal6 DA uptake assay.
Current methods combined with standard immunoblotting protocols provide the researcher with relevant tools to characterize the DAergic system.

Synaptosomal dopamine uptake and high-performance liquid chromatography analysis represent experimental tools to investigate dopamine homeostasis in mice by assessing the function of the dopamine transporter and levels of dopamine in striatal tissue, respectively. Here we present protocols to measure dopamine tissue content and assess the functionality of the dopamine transporter.

高速液体クロマトグラフィー分析とシナプトソーム ドーパミンによるマウスにおけるドーパミン作動性恒常性の評価

Dopamine (DA) is a modulatory neurotransmitter controlling motor activity, reward processes and cognitive function. Impairment of dopaminergic (DAergic) ...

An Objective and Reproducible Test of Olfactory Learning and Discrimination in Mice

Olfaction is the predominant sensory modality in mice and influences many important behaviors, including foraging, predator detection, mating, and parenting. Importantly, mice can be trained to associate novel odors with specific behavioral responses to provide insight into olfactory circuit function. This protocol details the procedure for training mice on a Go/No-Go operant learning task. In this approach, mice are trained on hundreds of automated trials daily for 2&#8211;4 weeks and can then be tested on novel Go/No-Go odor pairs to assess olfactory discrimination, or be used for studies on how odor learning alters the structure or function of the olfactory circuit. Additionally, the mouse olfactory bulb (OB) features ongoing integration of adult-born neurons. Interestingly, olfactory learning increases both the survival and synaptic connections of these adult-born neurons. Therefore, this protocol can be combined with other biochemical, electrophysiological, and imaging techniques to study learning and activity-dependent factors that mediate neuronal survival and plasticity.

Here, we train mice on an associative learning task to test odor discrimination. This protocol also allows for studies on learning-induced structural changes in the brain.

嗅覚学習とマウスにおける差別の客観的かつ再現性のあるテスト

Olfaction is the predominant sensory modality in mice and influences many important behaviors, including foraging, predator detection, mating, and ...

Transcranial Direct Current Stimulation (tDCS) in Mice

Transcranial direct current stimulation (tDCS) is a non-invasive neuromodulation technique proposed as an alternative or complementary treatment for several neuropsychiatric diseases. The biological effects of tDCS are not fully understood, which is in part explained due to the difficulty in obtaining human brain tissue. This protocol describes a tDCS mouse model that uses a chronically implanted electrode allowing the study of the long-lasting biological effects of tDCS. In this experimental model, tDCS changes the cortical gene expression and offers a prominent contribution to the understanding of the rationale for its therapeutic use.

Transcranial Direct Current Stimulation tDCS in Mice

Transcranial direct current stimulation (tDCS) is a therapeutic technique proposed to treat psychiatric diseases. An animal model is essential for understanding the specific biological alterations evoked by tDCS. This protocol describes a tDCS mouse model that uses a chronically implanted electrode.

マウスにおける経頭蓋直流電流刺激 (tDCS)

Transcranial direct current stimulation (tDCS) is a non-invasive neuromodulation technique proposed as an alternative or complementary treatment for ...

Quantitative Measurement of Intrathecally Synthesized Proteins in Mice

Cerebrospinal fluid (CSF), a fluid found in the brain and the spinal cord, is of great importance to both basic and clinical science. The analysis of the CSF protein composition delivers crucial information in basic neuroscience research as well as neurological diseases. One caveat is that proteins measured in CSF may derive from both intrathecal synthesis and transudation from serum, and protein analysis of CSF can only determine the sum of these two components. To discriminate between protein transudation from the blood and intrathecally produced proteins in animal models as well as in humans, CSF protein profiling measurements using conventional protein analysis tools must include the calculation of the albumin CSF/serum quotient (Qalbumin), a marker of the integrity of the blood-brain interface (BBI), and the protein index (Qprotein/Qalbumin), an estimate of intrathecal protein synthesis. This protocol illustrates the entire procedure, from CSF and blood collection to quotients and indices calculations, for the quantitative measurement of intrathecal protein synthesis and BBI impairment in mouse models of neurological disorders.

Elevated spinal fluid protein levels can either be the result of diffusion of plasma protein across an altered blood-brain barrier or intrathecal synthesis. An optimized testing protocol is presented in this article that helps to discriminate both cases and provides quantitative measurements of intrathecally synthesized proteins.

マウスにおけるインテコール内合成タンパク質の定量測定

Cerebrospinal fluid (CSF), a fluid found in the brain and the spinal cord, is of great importance to both basic and clinical science. The analysis of the ...

Electrocorticographic Recording of Cerebral Cortex Areas Manipulated Using an Adeno-Associated Virus Targeting Cofilin in Mice

The use of electrocorticographic (ECoG) recordings in rodents is relevant to sleep research and to the study of a wide range of neurological conditions. Adeno-associated viruses (AAVs) are increasingly used to improve understanding of brain circuits and their functions. The AAV-mediated manipulation of specific cell populations and/or of precise molecular components has been tremendously useful to identify new sleep regulatory circuits/molecules and key proteins contributing to the adverse effects of sleep loss. For instance, inhibiting activity of the filamentous actin-severing protein&#160;cofilin&#160;using AAV prevents sleep deprivation-induced memory impairment. Here, a protocol is&#160;described that combines the manipulation of cofilin function in a cerebral cortex area with the recording of ECoG activity to examine whether cortical cofilin modulates the wakefulness and sleep ECoG signals. AAV injection is performed during the same surgical procedure as the implantation of ECoG and electromyographic (EMG) electrodes in adult male and female mice. Mice are anesthetized, and their heads are shaved. After skin cleaning and incision, stereotaxic coordinates of the motor cortex are determined, and the skull is pierced at this location. A cannula prefilled with an AAV expressing cofilinS3D, an inactive form of cofilin, is slowly positioned in the cortical tissue. After AAV infusion, gold-covered screws (ECoG electrodes) are screwed through the skull and cemented to the skull with gold wires inserted in the neck muscles (EMG electrodes). The animals are allowed three weeks to recover and to ensure sufficient expression of cofilinS3D. The infected area and cell type are verified using immunohistochemistry, and the ECoG is analyzed using visual identification of vigilance states and spectral analysis. In summary, this combined methodological approach allows the investigation of the precise contribution of molecular components regulating neuronal morphology and connectivity to the regulation of synchronized cerebral cortex activity during wakefulness and sleep.

This article describes a protocol for the manipulation of molecular targets in the cerebral cortex using adeno-associated viruses and for monitoring the effects of this manipulation during wakefulness and sleep using electrocorticographic recordings.

マウスにおけるコフィリンを標的とするアデノ関連ウイルスを用いて操作した大脳皮質領域の電気コルチコチコ

The use of electrocorticographic (ECoG) recordings in rodents is relevant to sleep research and to the study of a wide range of neurological conditions. ...

マウスの運動皮質における細胞外電気生理学的記録

Multichannel Extracellular Recording in Freely Moving Mice

The protocol aims to uncover the properties of neuronal firing and network local field potentials (LFPs) in behaving mice carrying out specific tasks by correlating the electrophysiological signals with spontaneous and/or specific behavior. This technique represents a valuable tool in studying the neuronal network activity underlying these behaviors. The article provides a detailed and complete procedure for electrode implantation and consequent extracellular recording in free-moving conscious mice. The study includes a detailed method for implanting the microelectrode arrays, capturing the LFP and neuronal spiking signals in the motor cortex (MC) using a multichannel system, and the subsequent offline data analysis. The advantage of multichannel recording in conscious animals is that a greater number of spiking neurons and neuronal subtypes can be obtained and compared, which allows the evaluation of the relationship between a specific behavior and the associated electrophysiological signals. Notably, the multichannel extracellular recording technique and the data analysis procedure described in the present study can be applied to other brain areas when conducting experiments in behaving mice.

著者スポットライト:自由に動くマウスのニューロン活動を研究するためのマルチチャンネル細胞外記録の進歩 

The protocol describes the methodology of extracellular recording in the motor cortex (MC) to reveal extracellular electrophysiological properties in freely moving conscious mice, as well as the data analysis of local field potentials (LFPs) and spikes, which is useful for evaluating the network neural activity underlying behaviors of interest.

自由に動くマウスにおけるマルチチャンネル細胞外記録

The protocol aims to uncover the properties of neuronal firing and network local field potentials (LFPs) in behaving mice carrying out specific tasks by ...