ヘルムート ・ ショイレーベ ホーフグート Neumühle で豚舎で研究を実施する機会のために感謝したいと思います。

すべてのプロシージャ (#23177-07/G10-1-010/G 15-15-011) ローカル倫理委員会で承認された、欧州およびドイツの国民の規則 (欧州社会理事会指令、86/609/ECC が続いたTierschutzgesetz)。
すべての動物手術は医療センターのヨハネス ・ グーテンベルク大学マインツ アニマル ・ ケア委員会の規則に従って。
1. セットアップ
<ol><li>実験前に回線のノイズを確認し、アンテナとセットアップの適切な場所を見つけます。回線のノイズが 60 か 50 Hz の正弦波として表示されます。 注意: アンテナと送信機と受信機間の距離特にの配置は、システムの伝送強さによって決まります。ここで使用されるシステムは、調整可能です。それは比較的低消費電力、約 3 の m 伝送可能にされました。また、豚舎で金属フェンスできる信号を湿らせ、干渉を引き起こします。この場合、金属製のケージ内にアンテナを配置する必要です。</li>
	<li>ケーブル ドラムを使用して電源をセットアップします。使用されている特定のテレメトリ システムのラップトップ、レシーバー ユニット、アナログ-デジタル変換 (必要な場合) に接続します。 注: ここで使用される遠隔測定システムは、受信機にデジタル データを送信しました。これは、他のシステムの異なる場合があります。</li>
	<li>ミキシング ブロックと同様、電極、接着剤、綿棒とワイプを別のテーブルに配置します。</li>
	<li>短いケーブルの電極を準備します。電極をカットし、動物のサイズに応じて、可能な限り短い長さを再度半田付け。ケーブル必要がありますデータの転送、遠隔の脳波ユニットと頭の上の目的の記録の位置を接続するのに十分な長さ。長すぎるケーブルを反跳し、皮膚接着剤シリコーン ・ エラストマーで覆われている必要があります。反跳する必要があります長いケーブルは、大きくて重いシリコーン修正パッチを作る。</li>
</ol>2. 子豚
<ol><li>胸部、脚でつかんで子豚をキャッチします。それを保持し、排便や排尿に注意してください。</li>
	<li>必要に応じて、番号を持つ子豚をマークします。</li>
	<li>タオルで子豚をラップします。子豚が静まるでしょう。子豚の過熱の注意してください。</li>
	<li>体や前腕で片手で子豚を保持します。鼻を保持するために他の手を使用します。子豚の過熱に注意してください、それは正常な呼吸に無料かどうかを確認します。</li>
</ol>3. 電極
<ol><li>電極を接続する 2 番目の人があります。</li>
	<li>水やエタノールで汚れから肌をクリーンアップします。必要な場合は、頭部を剃る。</li>
	<li>研磨の脳波ゲルと q-ティップ、死んだ皮膚細胞を削除します。その後研磨ジェルを取り外します。また、サンドペーパーを使用します。</li>
	<li>目的の場所に自己粘着性の電極を修正します。(耳) 間小脳上接地電極を配置と鼻の上参照電極。目的の場所に記録電極を配置します。 注: この場合、単極記録だったため、実行参照は中立的な立場 (鼻部) に置かれました。ない標準化されたシステム利用できるある子豚の今まで。ここでは、頭頂部記録位置は、左右の脳半球に (目と耳) の間使用されました。</li>
	<li>テレメトリ ユニットにケーブルを接続します。ユニットの電源を入れます。テレメトリ システムの使用によってこれは磁気スイッチまたは無線周波数のウェイク アップ信号かもしれません。</li>
	<li>テレメトリ ユニットをカバーし、すべてのケーブル 2 成分皮膚接着シリコーンゴムを使用だけでなく、すべての電極 (材料の表を参照してください)。両方のコンポーネントの同量を混合することによって硬化時間が 1 分の目の範囲になり、まつげは、ゴムに覆われるべきであるないです。</li>
	<li>シリコーン ゴムが完全に硬化するまで待ちます。</li>
	<li>豚小屋に戻って子豚を配置します。</li>
	<li>ピグレット (数分) の時間の長い期間にわたって不快の徴候を示しているかどうかを確認します。</li>
</ol>4. 測定
<ol><li>通常の 30 後子豚が治りました (餌、遊んで、寝ている)、その兄弟とその動作を同期を開始するまでを待つ s (図 1)。</li>
	<li>必要な場合は、睡眠段階を待ちます。録音時間は、特定の科学的問題に依存します。ここでは、10 分のレコーディング セッションが使用されました。</li>
	<li>テレメトリ ユニットは、他の 2 以上の子豚で覆われている、信号が受信機の低すぎる可能性があります。彼らは上に眠っている場合、子豚を離れて軽く押します。の種をまくに注意してくださいそれは積極的に反応する可能性があります。</li>
	<li>データ集録ソフトウェアが記録を開始 (材料の表を参照してください)。</li>
</ol>5. 仕上げ加工
<ol><li>(通常数時間) 記録をキャッチ後子豚再びとして記載されている手順 2 で。の種をまくに注意してくださいそれは積極的に反応する可能性があります。</li>
	<li>1 つのエッジにシリコン ラバーをそっと持ち上げます。その後、電極およびテレメトリ ユニットを含むシリコーン ゴムのパッチ全体を削除します。子豚の目に注意します。</li>
	<li>豚小屋に戻って子豚を配置します。</li>
</ol>

<ol>
	<li>Conrad, M. S., Sutton, B. P., Dilger, R. N., Johnson, R. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=An+in+vivo+three-dimensional+magnetic+resonance+imaging-based+averaged+brain+collection+of+the+neonatal+piglet+(Sus+scrofa).">An in vivo three-dimensional magnetic resonance imaging-based averaged brain collection of the neonatal piglet (Sus scrofa).</a> PLoS ONE. 9 (9), e107650 (2014).</li><li>de Camp, N. V., Hense, F., Lecher, B., Scheu, H., Bergeler, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Models+for+preterm+cortical+development+using+non+invasive+clinical+EEG.">Models for preterm cortical development using non invasive clinical EEG.</a> Translational Neuroscience. 8, 211-224 (2017).</li><li>Lapray, D., Bergeler, J., Dupont, E., Thews, O., Luhmann, H. 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著者が明らかに何もありません。

プロトコルの重要なステップは、十分な皮膚の接触電極、特に接地電極低ノイズで安定した録音を達成するためにです。さらに、豚は非常に機敏なので、シリコーン ゴム電極とテレメトリ ユニットを保護するためにシステム全体をカバーすることが重要です。さらに、スラット床で安定に実験を行う場合は、小型のデバイスまたはコネクタに注意します。
自己粘着ゲル...

子豚は、神経科学1新興モデル システムです。橋渡し研究を強化するために、我々 は気ままな子豚2 (図 1および図 2) から非侵襲的、臨床脳波を記録する方法を発明しました。脳波、脳の成熟に関連する脳波パターンについての並進使用の 2 つの前提条件は、非侵襲的な方法論、臨床設定と鎮静剤や麻酔の禁欲に匹敵します。自己粘着性電極との組み合わせで 1 ch テレメトリー システム3その後約 5 分で修正できる、豚を処理プロシージャから迅速にリカバリし、餌や睡眠、その他の行動を同期させる豚と雌豚。
にもかかわらず、すでに鎮静動物4から非侵襲的な脳波を使用する試みがある、動物から脳波研究のほとんどを侵襲的アプローチを行っています。これらのメソッドは、炎症性プロセス注入電極5,6周りに関する副作用を持っている、ほとんどの場合、彼らは注入脳波システムの外部コンポーネントによる動物の社会的分離を必要と。したがって、これらのデータを臨床コンテキストの翻訳は難しいです。トランスレーショナル アプローチの必要性は、実際には、それはまだ知られていない臨床、非侵襲的な脳波7で「普通の」脳成熟皮質発育初期の表現方法によって明らかになっています。この知識のギャップは早産から脳波に関連付けられている技術的な課題による8の赤ちゃん。動物のモデル システムでは、ほとんどの動物は、人間の大脳皮質発達9と比較すると「早産脳」と生まれているので初期皮質発達のパターンいますアクセス。皮質の開発は、2種の保存されたパターンがある他に最近示されている preterm 赤ちゃんから脳波では、個々 の臨床結果を後の人生10,11時に予測できるも。ここで説明する方法は、大隅の並進の側面の便利です。

<table border="0" cellpadding="0" cellspacing="0" width="867">
	<tbody>
		<tr height="63">
			<td height="63" style="height:63px;width:216px;">Disposable adhesive 
			surface silver/silver chloride electrodes</td>
			<td style="width:309px;">Spes 
			Medica S.r.l., Genova, Italy</td>
			<td style="width:119px;"></td>
			<td style="width:223px;">Self adhesive hydrogel electrode</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Abralyt HiCl</td>
			<td style="width:309px;">Easycap GmbH</td>
			<td style="width:119px;"></td>
			<td>Abrasive cream</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Body Double fast</td>
			<td style="width:309px;">Smooth On Inc.</td>
			<td style="width:119px;"></td>
			<td>Skin adhesive silicone</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Telemetry system</td>
			<td style="width:309px;"></td>
			<td style="width:119px;"></td>
			<td>Internal development</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Picolog 1216</td>
			<td style="width:309px;">Pico Technology</td>
			<td style="width:119px;"></td>
			<td>AD converter</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Laptop</td>
			<td style="width:309px;">Panasonic</td>
			<td style="width:119px;"></td>
			<td>Rugged laptop</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Receiver</td>
			<td style="width:309px;"></td>
			<td style="width:119px;"></td>
			<td>Internal development</td>
		</tr>
	</tbody>
</table>

noninvasive eeg recordings from freely moving piglets

メソッドは、直接豚舎で豚を自由に移動から高品質脳波の記録を許可します。標準的な自己粘着ゲル電極との組み合わせで 1 ch 遠隔脳波システムを使用します。豚は、鎮静剤を使用せず落ち着いて。豚舎に彼らのリリース後、子豚は正常に動作-彼らは飲む、睡眠、兄弟と同じサイクルで。自分の睡眠段階は、脳波記録に使用されます。

関連付けられている、非-レム睡眠, スピンドル バーストや子豚 (図 1および図 2) を自由に移動するから、デルタのブラシのような典型的な脳波を記録することができました。代表的なパターンに興味を持っていた主の間にレム睡眠は、以外の非常に低振幅とレム様睡眠12の段階もされている (<strong class="xfi...

Watch this Scientific Journal Video about Noninvasive EEG Recordings from Freely Moving Piglets at JoVE.com

Noninvasive EEG Recordings from Freely Moving Piglets

ここでは、レコード遠隔脳波に直接鎮静剤を使わない豚舎で豚を自由に移動からのプロトコルを提案、バースト典型的な脳波パターンを記録中にレム睡眠は、以外の主軸のようすることが可能。

子豚を自由に移動するから非侵襲的脳波

The method allows the recording of high-quality electroencephalograms (EEGs) from freely moving piglets directly in the pigpen. We use a one-channel ...

noninvasive-eeg-recordings-from-freely-moving-piglets

Research

JoVE Journal

Neuroscience

7.2K ビュー.  Free University Berlin. ここでは、レコード遠隔脳波に直接鎮静剤を使わない豚舎で豚を自由に移動からのプロトコルを提案、バースト典型的な脳波パターンを記録中にレム睡眠は、以外の主軸のようすることが可能。

ビデオ: Noninvasive EEG Recordings from Freely Moving Piglets

High-density EEG Recordings of the Freely Moving Mice using Polyimide-based Microelectrode

Electroencephalogram (EEG) indicates the averaged electrical activity of the neuronal populations on a large-scale level. It is widely utilized as a noninvasive brain monitoring tool in cognitive neuroscience as well as a diagnostic tool for epilepsy and sleep disorders in neurology. However, the underlying mechanism of EEG rhythm generation is still under the veil. Recently introduced polyimide-based microelectrode (PBM-array) for high resolution mouse EEG1 is one of the trials to answer the neurophysiological questions on EEG signals based on a rich genetic resource that the mouse model contains for the analysis of complex EEG generation process. This application of nanofabricated PBM-array to mouse skull is an efficient tool for collecting large-scale brain activity of transgenic mice and accommodates to identify the neural correlates to certain EEG rhythms in conjunction with behavior. However its ultra-thin thickness and bifurcated structure cause a trouble in handling and implantation of PBM-array. In the presented video, the preparation and surgery steps for the implantation of PBM-array on a mouse skull are described step by step. Handling and surgery tips to help researchers succeed in implantation are also provided.

In this article, we described the surgery procedure and handling tips for implantation of ultra-thin polyimide-based microelectrode array (PBM-array) on the mouse skull for acquisition of high-density encephalography (EEG) in a mouse model.

ポリイミドベースのマイクロ電極を用いて自由に動くマウスの高密度脳波録音

Electroencephalogram (EEG) indicates the averaged electrical activity of the neuronal populations on a large-scale level. It is widely utilized as a ...

神経科学

Patch Clamp Recordings from Embryonic Zebrafish Mauthner Cells

Mauthner cells (M-cells) are large reticulospinal neurons located in the hindbrain of teleost fish. They are key neurons involved in a characteristic behavior known as the C-start or escape response that occurs when the organism perceives a threat. The M-cell has been extensively studied in adult goldfish where it has been shown to receive a wide range of excitatory, inhibitory and neuromodulatory signals1. We have been examining M-cell activity in embryonic zebrafish in order to study aspects of synaptic development in a vertebrate preparation. In the late 1990s Ali and colleagues developed a preparation for patch clamp recording from M-cells in zebrafish embryos, in which the CNS was largely intact2,3,4. The objective at that time was to record synaptic activity from hindbrain neurons, spinal cord neurons and trunk skeletal muscle while maintaining functional synaptic connections within an intact brain-spinal cord preparation. This preparation is still used in our laboratory today. To examine the mechanisms underlying developmental synaptic plasticity, we record excitatory (AMPA and NMDA-mediated)5,6 and inhibitory (GABA and glycine) synaptic currents from developing M-cells. Importantly, this unique preparation allows us to return to the same cell (M-cell) from preparation to preparation to carefully examine synaptic plasticity and neuro-development in an embryonic organism. The benefits provided by this preparation include 1) intact, functional synaptic connections onto the M-cell, 2) relatively inexpensive preparations, 3) a large supply of readily available embryos 4) the ability to return to the same cell type (i.e. M-cell) in every preparation, so that synaptic development at the level of an individual cell can be examined from fish to fish, and 5) imaging of whole preparations due to the transparent nature of the embryos.

We have developed an intact brain-spinal cord preparation to record and monitor electrical activity via patch clamp recording from the Mauthner neurons and other reticulospinal cells in zebrafish embryos. Thus, we are able to record excitatory and inhibitory synaptic currents, voltage-gated channel activity and action potentials from key neurons in a developing embryo.

胚ゼブラフィッシュマウスナー細胞からパッチクランプ記録

Mauthner cells (M-cells) are large  reticulospinal neurons located in the hindbrain of teleost fish. They are key  neurons involved in a characteristic ...

Recording Single Neurons' Action Potentials from Freely Moving Pigeons Across Three Stages of Learning

While the subject of learning has attracted immense interest from both behavioral and neural scientists, only relatively few investigators have observed single-neuron activity while animals are acquiring an operantly conditioned response, or when that response is extinguished. But even in these cases, observation periods usually encompass only a single stage of learning, i.e. acquisition or extinction, but not both (exceptions include protocols employing reversal learning; see Bingman&#160;et al.1 for an example). However, acquisition and extinction entail different learning mechanisms and are therefore expected to be accompanied by different types and/or loci of neural plasticity.
Accordingly, we developed a behavioral paradigm which institutes three stages of learning in a single behavioral session and which is well suited for the simultaneous recording of single neurons&#39; action potentials. Animals are trained on a single-interval forced choice task which requires mapping each of two possible choice responses to the presentation of different novel visual stimuli (acquisition). After having reached a predefined performance criterion, one of the two choice responses is no longer reinforced (extinction). Following a certain decrement in performance level, correct responses are reinforced again (reacquisition). By using a new set of stimuli in every session, animals can undergo the acquisition-extinction-reacquisition process repeatedly. Because all three stages of learning occur in a single behavioral session, the paradigm is ideal for the simultaneous observation of the spiking output of multiple single neurons. We use pigeons as model systems, but the task can easily be adapted to any other species capable of conditioned discrimination learning.

Learning new stimulus-response associations engages a wide range of neural processes which are ultimately reflected in changing spike output of individual neurons. Here we describe a behavioral protocol allowing for the continuous registration of single-neuron activity while animals acquire, extinguish, and reacquire a conditioned response within a single experimental session.

自由に学習の三つの段階を越えハトを移動する単一ニューロン」の活動電位を記録する

While the subject of learning has attracted immense interest from both behavioral and neural scientists, only relatively few investigators have observed ...

A Novel Behavioral Assay to Investigate Gustatory Responses of Individual, Freely-moving Bumble Bees (Bombus terrestris)

Generalist pollinators like the buff-tailed bumble bee, Bombus terrestris, encounter both nutrients and toxins in the floral nectar they collect from flowering plants. Only a few studies have described the gustatory responses of bees toward toxins in food, and these experiments have mainly used the proboscis extension response on restrained honey bees. Here, a new behavioral assay is presented for measuring the feeding responses of freely-moving, individual worker bumble bees to nutrients and toxins. This assay measures the amount of solution ingested by each bumble bee and identifies how tastants in food influence the microstructure of the feeding behavior.
The solutions are presented in a microcapillary tube to individual bumble bees that have been previously starved for 2-4 hr. The behavior is captured on digital video. The fine structure of the feeding behavior is analyzed by continuously scoring the position of the proboscis (mouthparts) from video recordings using event logging software. The position of the proboscis is defined by three different behavioral categories: (1) proboscis is extended and in contact with the solution, (2) proboscis is extended but not in contact with the solution and (3) proboscis is stowed under the head. Furthermore the speed of the proboscis retracting away from the solution is also estimated.
In the present assay the volume of solution consumed, the number of feeding bouts, the duration of the feeding bouts and the speed of the proboscis retraction after the first contact is used to evaluate the phagostimulatory or the deterrent activity of the compounds tested.
This new taste assay will allow researchers to measure how compounds found in nectar influence the feeding behavior of bees and will also be useful to pollination biologists, toxicologists and neuroethologists studying the bumble bee&#39;s taste system.

A Novel Behavioral Assay to Investigate Gustatory Responses of Individual, Freely-moving Bumble Bees Bombus terrestris

A novel behavioral assay is described for investigating the short term gustatory responses of the mouthparts of freely-moving bumble bees (Bombus terrestris) toward nutrients and toxins in solution.

小説行動アッセイ個人の味覚応答を調査するために、自由に移動バンブル蜂（<em&gt;セイヨウオオマルハナバチ</em&gt;）

Generalist pollinators like the buff-tailed bumble bee, Bombus terrestris, encounter both nutrients and toxins in the floral nectar they collect from ...

Recording EEG in Freely Moving Neonatal Rats Using a Novel Method

EEG is a useful method to detect electrical activity in the brain. Moreover, it is a widely used diagnostic tool for various neurological conditions, such as epilepsy and neurodegenerative disorders. However, it is technically difficult to obtain EEG recordings in neonates as it requires specialized handling and great care. Here, we present a novel method to record EEG in neonatal rat pups (P8-P15). We designed a simple and reliable electrode using computer pin loci; it can be easily implanted into the skull of a rat pup to record high-quality EEG signals in the normal and epileptic brain. Pups were given an intraperitoneal (i.p.) injection of the neurotoxin kainic acid (KA) to induce epileptic seizures. The surgical implantation performed in this procedure is less expensive than other EEG procedures for neonates. This method allows one to record high-quality and stable EEG signals for more than 1 week. Furthermore, this procedure can also be applied to adult rats and mice to study epilepsy or other neurological disorders.

Here, we introduce a novel technique designed to record electroencephalography (EEG) in freely moving neonatal epileptic pups and describe its procedures, features, and applications. This method allows one to record EEG for more than 1 week.

新しい方法を用いた自由に動く新生児ラットの脳波記録

EEG is a useful method to detect electrical activity in the brain. Moreover, it is a widely used diagnostic tool for various neurological conditions, such ...

Visual Evoked Potential Recordings in Mice Using a Dry Non-invasive Multi-channel Scalp EEG Sensor

For scalp EEG research environments with laboratory mice, we designed a dry-type 16 channel EEG sensor which is non-invasive, deformable, and re-usable because of the plunger-spring-barrel structural facet and mechanical strengths resulting from metal materials. The whole process for acquiring the VEP responses in vivo from a mouse consists of four steps: (1) sensor assembly, (2) animal preparation, (3) VEP measurement, and (4) signal processing. This paper presents representative measurements of VEP responses from multiple mice with a submicro-voltage signal resolution and sub-hundred millisecond temporal resolution. Although the proposed method is safer and more convenient compared to other previously reported animal EEG acquiring methods, there are remaining issues including how to enhance the signal-to-noise ratio and how to apply this technique with freely moving animals. The proposed method utilizes easily available resources and shows a repetitive VEP response with a satisfactory signal quality. Therefore, this method could be utilized for longitudinal experimental studies and reliable translational research exploiting non-invasive paradigms.

We designed a dry-type 16 channel EEG sensor which is non-invasive, deformable, and re-usable. This paper describes the whole process from manufacturing the proposed EEG electrode to signal processing of visual evoked potential (VEP) signals measured on a mouse scalp using a dry non-invasive multi-channel EEG sensor.

誘発電位録画ドライの非侵襲的なマルチ チャネルを使用してマウスの頭皮脳波センサー

For scalp EEG research environments with laboratory mice, we designed a dry-type 16 channel EEG sensor which is non-invasive, deformable, and re-usable ...

Simultaneous Recordings of Cortical Local Field Potentials, Electrocardiogram, Electromyogram, and Breathing Rhythm from a Freely Moving Rat

Monitoring the physiological dynamics of the brain and peripheral tissues is necessary for addressing a number of questions about how the brain controls body functions and internal organ rhythms when animals are exposed to emotional challenges and changes in their living environments. In general experiments, signals from different organs, such as the brain and the heart, are recorded by independent recording systems that require multiple recording devices and different procedures for processing the data files. This study describes a new method that can simultaneously monitor electrical biosignals, including tens of local field potentials in multiple brain regions, electrocardiograms that represent the cardiac rhythm, electromyograms that represent awake/sleep-related muscle contraction, and breathing signals, in a freely moving rat. The recording configuration of this method is based on a conventional micro-drive array for cortical local field potential recordings in which tens of electrodes are accommodated, and the signals obtained from these electrodes are integrated into a single electrical board mounted on the animal's head. Here, this recording system was improved so that signals from the peripheral organs are also transferred to an electrical interface board. In a single surgery, electrodes are first separately implanted into the appropriate body parts and the target brain areas. The open ends of all of these electrodes are then soldered to individual channels of the electrical board above the animal's head so that all of the signals can be integrated into the single electrical board. Connecting this board to a recording device allows for the collection of all of the signals into a single device, which reduces experimental costs and simplifies data processing, because all data can be handled in the same data file. This technique will aid the understanding of the neurophysiological correlates of the associations between central and peripheral organs.

This study introduces a method for the simultaneous recording of local field potentials in the brain, electrocardiograms, electromyograms, and breathing signals of a freely moving rat. This technique, which reduces experimental costs and simplifies data analysis, will contribute to the understanding of the interactions between the brain and peripheral organs.

皮質局所電界電位、心電図、筋電図、呼吸リズム自由行動ラットからの同時録音

Monitoring the physiological dynamics of the brain and peripheral tissues is necessary for addressing a number of questions about how the brain controls ...

Microdialysis of Excitatory Amino Acids During EEG Recordings in Freely Moving Rats

Microdialysis is a well-established neuroscience technique that correlates the changes of neurologically active substances diffusing into the brain interstitial space with the behavior and/or with the specific outcome of a pathology (e.g., seizures for epilepsy). When studying epilepsy, the microdialysis technique is often combined with short-term or even long-term video-electroencephalography (EEG) monitoring to assess spontaneous seizure frequency, severity, progression and clustering. The combined microdialysis-EEG is based on the use of several methods and instruments. Here, we performed in vivo microdialysis and continuous video-EEG recording to monitor glutamate and aspartate outflow over time, in different phases of the natural history of epilepsy in a rat model. This combined approach allows the pairing of changes in the neurotransmitter release with specific stages of the disease development and progression. The amino acid concentration in the dialysate was determined by liquid chromatography. Here, we describe the methods and outline the principal precautionary measures one should take during in vivo microdialysis-EEG, with particular attention to the stereotaxic surgery, basal and high potassium stimulation during microdialysis, depth electrode EEG recording and high-performance liquid chromatography analysis of aspartate and glutamate in the dialysate. This approach may be adapted to test a variety of drug or disease induced changes of the physiological concentrations of aspartate and glutamate in the brain. Depending on the availability of an appropriate analytical assay, it may be further used to test different soluble molecules when employing EEG recording at the same time.

Here, we describe a method for in vivo microdialysis to analyze aspartate and glutamate release in the ventral hippocampus of epileptic and non-epileptic rats, in combination with EEG recordings. Extracellular concentrations of aspartate and glutamate may be correlated with the different phases of the disease.

自由行動ラットの脳波記録中に興奮性アミノ酸の透析

Microdialysis is a well-established neuroscience technique that correlates the changes of neurologically active substances diffusing into the brain ...

Long-term Sensory Conflict in Freely Behaving Mice

Long-term sensory conflict protocols are a valuable means of studying motor learning. The presented protocol produces a persistent sensory conflict for experiments aimed at studying long-term learning in mice. By permanently wearing a device fixed on their heads, mice are continuously exposed to a sensory mismatch between visual and vestibular inputs while freely moving in home cages. Therefore, this protocol readily enables the study of the visual system and multisensory interactions over an extended timeframe that would not be accessible otherwise. In addition to lowering the experimental costs of long-term sensory learning in naturally behaving mice, this approach accommodates the combination of in vivo and in vitro experiments. In the reported example, video-oculography is performed to quantify the vestibulo-ocular reflex (VOR) and optokinetic reflex (OKR) before and after learning. Mice exposed to this long-term sensory conflict between visual and vestibular inputs presented a strong VOR gain decrease but exhibited few OKR changes. Detailed steps of device assembly, animal care, and reflex measurements are hereby reported.

The presented protocol produces a persistent sensory conflict for experiments aimed at studying long-term learning. By permanently wearing a fixed device on their heads, mice are continuously exposed to a sensory mismatch between visual and vestibular inputs while freely moving in home cages.

自由に行動するマウスで長期的な感覚の衝突

Long-term sensory conflict protocols are a valuable means of studying motor learning. The presented protocol produces a persistent sensory conflict for ...

Hybrid Microdrive System with Recoverable Opto-Silicon Probe and Tetrode for Dual-Site High Density Recording in Freely Moving Mice

Multi-regional neural recordings can provide crucial information to understanding fine-timescale interactions between multiple brain regions. However, conventional microdrive designs often only allow use of one type of electrode to record from single or multiple regions, limiting the yield of single-unit or depth profile recordings. It also often limits the ability to combine electrode recordings with optogenetic tools to target pathway and/or cell type specific activity. Presented here is a hybrid microdrive array for freely moving mice to optimize yield and a description of its fabrication and reuse of the microdrive array. The current design employs nine tetrodes and one opto-silicon probe implanted in two different brain areas simultaneously in freely moving mice. The tetrodes and the opto-silicon probe are independently adjustable along the dorsoventral axis in the brain to maximize the yield of unit and oscillatory activities. This microdrive array also incorporates a set-up for light, mediating optogenetic manipulation to investigate the regional- or cell type-specific responses and functions of long-range neural circuits. In addition, the opto-silicon probe can be safely recovered and reused after each experiment. Because the microdrive array consists of 3D-printed parts, the design of microdrives can be easily modified to accommodate various settings. First described is the design of the microdrive array and how to attach the optical fiber to a silicon probe for optogenetics experiments, followed by fabrication of the tetrode bundle and implantation of the array into a mouse brain. The recording of local field potentials and unit spiking combined with optogenetic stimulation also demonstrate feasibility of the microdrive array system in freely moving mice.

This protocol describes the construction of a hybrid microdrive array that allows implantation of nine independently adjustable tetrodes and one adjustable opto-silicon probe in two brain regions in freely moving mice. Also demonstrated is a method for safely recovering and reusing the opto-silicon probe for multiple purposes.

自由に動くマウスの二重部位高密度記録のための回収可能なオプトシリコンプローブとテトロードを備えたハイブリッドマイクロドライブシステム

Multi-regional neural recordings can provide crucial information to understanding fine-timescale interactions between multiple brain regions. However, ...