この作品は、研こんにちはによって支えられた (17 H 05939; 17 H 05551)、中富財団とスズケン記念財団。

動物を対象とする手順のすべては、ケアとの動物の使用のための NIH のガイドラインに従って行われました。
1. 統合のマイクロ ドライブ アレイの準備
<ol><li>1,2,3他の所で記述として皮質 LFP 録音用マイクロ ドライブ アレイを作成します。1.2 に従ってバイオフレックス ワイヤに接続されている心電図、筋電図、BR チャンネルとして用電極インタ フェース ボード (EIB) に少なくとも 6 のまま金属の穴を開きます。</li>
	<li>長さの 5.0 cm 皮ポリテトラフルオロ エチレン (PTFE) を 6 個セットにバイオフレックス カットワイヤ ~5.0 mm の長さを持つすべてのワイヤー部分の両端のコーティングに各開いている金属穴 (心電図、筋電図、BR チャンネル) の 1 つにワイヤー部分の一端を接続、。 金のピンと EIB。</li>
	<li>2 つの 5.0 cm 部分にエナメル線をカットします。各 EIB (図 3を参照してくださいも以前論文12,13) の地面/参照 (g ・ r) チャンネルにこれらのワイヤの一方の端をはんだ付けします。</li>
	<li>心電図用電極、2 つの 16 cm 部分にバイオフレックス カットワイヤを備える。~5.0 mm 片側 (短い方の端)、もう一方の端 (長い方の端) 〜 15 mm の長さでこれらのワイヤーの部分の両端の PTFE コーティングをはがします。</li>
	<li>2.0 mm の直径をワイヤー リングを形成すると、ワイヤの長い方の端を曲げ、はんだ付けによるリングの形状を修正します。</li>
	<li>筋電図電極の作製、バイオフレックス ワイヤーの長さの 8 cm の皮 ~5.0 ミリメートルの長さとこれらのワイヤー部分の両端から PTFE コーティングをオフで 2 枚にカットします。</li>
	<li>バイオフレックス ワイヤーの長さの 6.0 cm 皮 ~5.0 の長さとこれらのワイヤーの部分の両端のエナメル コーティングをオフで 2 個に切口の BR 電極、準備の mm はんだステンレス鋼ねじ (幹径の頭にこれらのワイヤーの作品のそれぞれの一方の端計: 1.0 mm、茎の長さ: 4.0 mm)。</li>
	<li>(Gr) の地面/参照電極の作製、カットの長さの 6.0 cm 皮 ~5.0 ミリメートルの長さとこれらのワイヤーの部分の両端のエナメル コーティングをオフで 2 個にエナメル線半田ステンレス st の頭にこれらのワイヤーの部分のそれぞれの一方の端うなぎネジ (茎の直径: 1.4 mm、茎の長さ: 3.0 mm)。</li>
	<li>ガスはすべての電極とステンレス ボルトを消毒、クリーンな空間でこれらを維持します。</li>
</ol>2. 心電図・筋電図電極の注入
メモ: は、滅菌手袋、オートクレーブ器を用いた無菌法による手術のすべての手順を実行します。切開の作成を含むすべての手順については、前に、70% エタノールで皮膚を殺菌し、手術用ドレープを使用して切開をカバーします。
<ol><li>麻酔を修正 (1.0 3.0% イソフルラン ガス) ラット フラット熱パッドの裏面。ブプレノルフィンを鎮痛薬として与えます。獣医軟膏を乾燥を防ぐためにラットの目に配置します。皮膚の表面をきれいに betadine を使用します。</li>
	<li>内側胸の地域で ~2.0 cm の切開を行います。胸の筋肉を分離することによって肋間筋を公開します。肋間筋に ECG 電極のリングを縫合します。</li>
	<li>熱パッドを動物の胃を修正します。背側頸部に ~1.0 cm の切開を行います。</li>
	<li>胸部切開皮下心電図電極を挿入します。背側頸部に両端をスライドさせ、首の切開からそれらを引き出します。胸部切開部を縫合します。</li>
	<li>筋電図電極を皮下首切開 ~2.0 センチメートルの長さのそれぞれの一方の端を挿入します。縫合によって首の筋肉の筋電図電極を修正します。</li>
</ol>3. 統合マイクロ ドライブ アレイと BR 電極の注入
<ol><li>脳定位固定装置のラットを修正します。頸部に目の間のポイントから正中線に沿って頭の上 ~3.0 cm の切開を行います。頭蓋骨を公開します。</li>
	<li>直径 0.7 〜 1.0 mm 11.0 mm 前方と 1 mm の嗅球上の 2 つの円形 craniotomies 両側に加える前高速ドリルで。インプラントの頭蓋骨の 2 BR 電極までネジ茎の先端が脳の表面に付着した.</li>
	<li>前に両側の 0.7 〜 1.0 mm 以上前頭皮質 2.7 mm 前方と 2.7 mm の直径を持つ 2 つの円形 craniotomies を加えます。ネジ幹の先端が脳の表面に接続されるまでは、頭蓋骨の 2 つの g/r 電極をインプラントします。</li>
	<li>周辺に大規模な開頭術 6 ~ 8 直径 1.0 mm の穴を作る。インプラント アンカーのネジ (茎の直径: 1.4 mm、茎の長さ: 3.0 mm) 頭蓋内。</li>
	<li>前に二国間の 3.8 mm 後方に, と海馬の上 ~2.0 mm、2.5 mm の直径の円形開を加えます。ドライブ アレイのカニューレ先端は大きな開頭の上にあるような統合のマイクロ ドライブ アレイを配置します。</li>
	<li>2 つのソリューション、すなわち~ 100 μ L とカニューレ先端と脳の表面との間のギャップ スペースを埋める。、0.5% (質量) アルギン酸ナトリウムと塩化カルシウム 10% (質量)。 注: このプロセス 〜 5 分、2 つのソリューションが頭蓋骨上で混合後透明なゲルを形成します。</li>
	<li>カニューレ、BR 電極、g ・ r 電極をカバーし、0.5 cm. の厚さで歯科用セメント アンカーねじが注意していないこの段階でセメントを BR と g/r 電極の開いた端部をカバーします。</li>
	<li>EIB に以前接続されていた個別のワイヤの先端に心電図、筋電図、BR と g/r 電極の開いた端部をはんだ付け (1.2 と 1.3 の手順を参照してください)。</li>
	<li>統合マイクロ ドライブ アレイと歯科用セメントとのすべての電極線の下の部分をカバーしてください。いるラット注入した後それらを傷つけることはできませんすべての電極ワイヤを完全に覆われていることを確認します。</li>
	<li>胸骨の横臥を維持するために十分な意識を取り戻した後、透明なプレキシ グラス ホーム cag に動物を戻り、水や食料を無料で独自にそれを維持します。手術後、抗生物質 (ゲンタマイシン) と動物を扱います。</li>
	<li>手術後に、毎日の観察と動物を監視します。正しく歩くことと彼らは、実験者はマイクロ ドライブ アレイを触れたときにないキーッか確認します。</li>
</ol>4.生体内での録音
注: すべての信号が増幅され、2 kHz でサンプリングと帯域通過フィルター (0.1 - 500 Hz) をユニット活動 (30 kHz でサンプリングとバンドパス フィルター (500-6 kHz)) を除く。
<ol><li>統合のマイクロ ドライブ アレイの EIB を録音デバイスのパッチアンプ用アダプターに接続します。</li>
	<li>手術後の数週間のネジを回して、管の四極管を進めます。管の四極管が対象脳領域に隣接する、安定した録音のための数日の期間の領域に、管の四極管を解決します。</li>
	<li>動物は記録室で自由に移動しながら電気信号を監視します。</li>
</ol>

<ol>
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S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Tetrode+technology:+advances+in+implantable+hardware,+neuroimaging,+and+data+analysis+techniques.">Tetrode technology: advances in implantable hardware, neuroimaging, and data analysis techniques.</a> J Neurosci Methods. 117 (2), 141-152 (2002).</li><li>Fenske, S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Comprehensive+multilevel+in+vivo+and+in+vitro+analysis+of+heart+rate+fluctuations+in+mice+by+ECG+telemetry+and+electrophysiology.">Comprehensive multilevel in vivo and in vitro analysis of heart rate fluctuations in mice by ECG telemetry and electrophysiology.</a> Nat Protoc. 11 (1), 61-86 (2016).</li><li>Rossi, S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+effect+of+aging+on+the+specialized+conducting+system:+a+telemetry+ECG+study+in+rats+over+a+6+month+period.">The effect of aging on the specialized conducting system: a telemetry ECG study in rats over a 6 month period.</a> PLoS One. 9 (11), 112697 (2014).</li><li>Cesarovic, N., Jirkof, P., Rettich, A., Arras, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Implantation+of+radiotelemetry+transmitters+yielding+data+on+ECG,+heart+rate,+core+body+temperature+and+activity+in+free-moving+laboratory+mice.">Implantation of radiotelemetry transmitters yielding data on ECG, heart rate, core body temperature and activity in free-moving laboratory mice.</a> JoVE. 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周辺のアクティビティ レベルと副の脳の変調を理解するため、逆に、大規模な複数の部位から電気の生体情報を同時にキャプチャする方法を記録が必要です。本研究の説明、手術は、脳の局所電界電位、心拍数、筋肉構造とに使用される記録システムに改善されてきた呼吸率の大きさを監視記録システム脳組織の細胞外の録音。このシステムは、統合マイクロ ドライブ配列の単一 EIB に脳と...

中枢神経系は様々 な環境変化に対応した身体の状態を制御し、このコントロールは、通常、心拍数、呼吸数と筋収縮の変化として表されます。ただし、いくつかの研究は、このような末梢の生理的要因が皮質活動に関連付けられている方法をテストしています。この問題を解決するには、は、中枢および末梢組織から電気の生体情報を監視するための大規模な記録方式が必要です。大脳皮質では、ローカル フィールド (LFP) シグナル細胞外皮質組織1,2,3に挿入される電極によって記録されます。ラットやマウスなどの小型哺乳類の皮質領域から複数の LFP 信号を同時に記録するには、多くの研究は、マイクロ ドライブと呼ばれる特注電極アセンブリの様々 なタイプを開発しました。従来のマイクロ ドライブは (これは通常管四極管) 電極、ネジや電極対応コア ボディと金属の穴に対応する電気的インタ フェース ボード (EIB) の中間部分に接続されている金属製のネジから成る(図 1図 2、および図 3) 電極の開いた端部を接続します。この電極アセンブリ多くの電極を数日から数週間、もちろん脳に挿入の深さをコントロールすることが可能でき、動物は様々 な挑戦として、神経活動を長期的な慢性的な記録の実施行動のタスク。末梢器官のハートビート信号はや4,エリア65,の心、周りに注入されている電極の組み合わせによって心電図 (Ecg) として記録され、骨格筋の信号を記録電極で筋電図 (筋電図) として筋組織7,8,9に挿入されました。単一ユニットの録音10,11で嗅球の電気信号と呼吸 (BR) との関係を調べた。従来の録音システムでさまざまな組織からこれらの信号によってキャプチャされた独立した録音デバイス追加実験システムが正確にこれら複数のデバイスを同期する必要があることを意味同時脳体信号の録音。このシステムは、この問題を克服するために開発されました。このシステムでは、心電図、筋電図、呼吸のリズムを反映する嗅球からの電気信号など末梢の器官から記録されたすべての電気信号が単一のマイクロ ドライブ アレイ1,2 に統合します。 ,3, ここで統合のマイクロ ドライブ アレイと呼ばれます。このシステムは 1 つだけマルチ チャンネル記録装置を必要とし、任意の従来のマイクロ ドライブの配列に適用されます。この手法の利点はそれにより信号のすべてが同じタイプのデータとして記録されるのでより便利なデータ処理のため特別なデバイスまたは複数のデバイスの録音時間に合わせてトリガー信号を必要としないことです。この手法は、中枢および末梢の器官間のアソシエーションの神経生理学的相関の理解に役立ちます。本技術に関連する手順をについて説明します、ラットから得られる代表的なデータセットを提示します。

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			<td></td>
		</tr>
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			<td height="40" style="height:40px;">Stainless steel screw M1.0&#215;4.0&#160;</td>
			<td>MonotaRO, Hyogo, Japan</td>
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			<td height="40" style="height:40px;width:307px;">Stainless steel screw M1.4&#215;3.0</td>
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simultaneous recordings of cortical local field potentials, electrocardiogram, electromyogram, and breathing rhythm from a freely moving rat

動物の感情的な課題と変化にさらされている脳コントロールで機能と内臓のリズムを体する方法に関する質問の数に対処するために必要な脳と末梢組織の生理的動態は、生活環境。一般に実験、脳や心臓など、さまざまな器官からの信号は、データ ファイルを処理するための複数の録音デバイスと異なる手順を必要とする独立した記録システムによって記録されます。本研究は、複数の脳の領域、心臓のリズムを表す心電図、目を覚ましを表す筋電図局所電界電位の数十を含む電気の生体情報を同時に監視できます新しい方法を記述/睡眠に関連する筋肉の収縮と自由行動ラットにおける信号を呼吸します。このメソッドの記録構成は皮質のローカル フィールド潜在的な記録電極の数十に対応し、これらの電極から得られる信号が 1 つに統合されている従来のマイクロ ドライブ配列に基づいてください。電気キャビネットは、動物の頭にマウント。ここでは、この記録システムの改良だったので末梢器官からの信号にも電気的インタ フェース ボードに転送。シングルの手術で電極が個別に適切な体の部分や対象脳領域に埋め込まれた最初。すべてこれらの電極のオープン エンドは、動物の頭の上の電気基板の個々 のチャンネルにはんだ付け、信号のすべてが単一の電気ボードに組み入れることができます。実験のコストを削減し、同じデータ ファイル内のすべてのデータを処理できるので、データ処理を簡素化する 1 つのデバイスにすべての信号のコレクション録音デバイスにこのボードを接続することができます。この手法は、中枢および末梢の器官間のアソシエーションの神経生理学的相関の理解に役立ちます。

このメソッドは、心拍数、呼吸のリズムと骨格筋収縮(図 1)は、脳の神経活動を表す複数の器官から生体電気信号を同時にキャプチャできます。図 4は、長方形の箱 (25 × 40 cm2) で自由に採餌は自由行動ラットからデータの代表的な記録を提供します。例のデータセットには、移動と静止状態との間の典?...

Watch this Scientific Journal Video about Simultaneous Recordings of Cortical Local Field Potentials, Electrocardiogram, Electromyogram, and Breathing Rhythm from a Freely Moving Rat at JoVE.com

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

本研究では、脳、心電図、筋電図、局所電界電位の同時計測と自由行動ラットの信号を呼吸するためのメソッドを紹介します。この技術は、実験のコストを削減し、データの分析を簡素化は、脳と末梢器官との間の相互作用の理解に貢献します。

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 ...

simultaneous-recordings-cortical-local-field-potentials

Research

JoVE Journal

Neuroscience

10.8K ビュー.  The University of Tokyo. 本研究では、脳、心電図、筋電図、局所電界電位の同時計測と自由行動ラットの信号を呼吸するためのメソッドを紹介します。この技術は、実験のコストを削減し、データの分析を簡素化は、脳と末梢器官との間の相互作用の理解に貢献します。

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

Single-unit In vivo Recordings from the Optic Chiasm of Rat

Information about the visual world is transmitted to the brain in sequences of action potentials in retinal ganglion cell axons that make up the optic nerve. In vivo recordings of ganglion cell spike trains in several animal models have revealed much of what is known about how the early visual system processes and encodes visual information. However, such recordings have been rare in one of the most common animal models, the rat, possibly owing to difficulty in detecting spikes fired by small diameter axons. The many retinal disease models involving rats motivate a need for characterizing the functional properties of ganglion cells without disturbing the eye, as with intraocular or in vitro recordings. Here, we demonstrate a method for recording ganglion cell spike trains from the optic chiasm of the anesthetized rat. We first show how to fabricate tungsten-in-glass electrodes that can pick up electrical activity from single ganglion cell axons in rat. The electrodes outperform all commercial ones that we have tried. We then illustrate our custom-designed stereotaxic system for in vivo visual neurophysiology experiments and our procedures for animal preparation and reliable and stable electrode placement in the optic chiasm.

Single-unit In vivo Recordings from the Optic Chiasm of Rat

Retinal ganglion cells transmit visual information from the eye to the brain with sequences of action potentials. Here, we demonstrate how to record the action potentials of single ganglion cells in vivo from anesthetized rats.

シングルユニット in vivoで</emラットの視交叉から>録音

Information about the visual world is transmitted to the brain in sequences of action potentials in retinal ganglion cell axons that make up the optic ...

神経科学

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 ...

Simultaneous Whole-cell Recordings from Photoreceptors and Second-order Neurons in an Amphibian Retinal Slice Preparation

One of the central tasks in retinal neuroscience is to understand the circuitry of retinal neurons and how those connections are responsible for shaping the signals transmitted to the brain. Photons are detected in the retina by rod and cone photoreceptors, which convert that energy into an electrical signal, transmitting it to other retinal neurons, where it is processed and communicated to central targets in the brain via the optic nerve. Important early insights into retinal circuitry and visual processing came from the histological studies of Cajal1,2 and, later, from electrophysiological recordings of the spiking activity of retinal ganglion cells - the output cells of the retina3,4.
A detailed understanding of visual processing in the retina requires an understanding of the signaling at each step in the pathway from photoreceptor to retinal ganglion cell. However, many retinal cell types are buried deep in the tissue and therefore relatively inaccessible for electrophysiological recording. This limitation can be overcome by working with vertical slices, in which cells residing within each of the retinal layers are clearly visible and accessible for electrophysiological recording.
Here, we describe a method for making vertical sections of retinas from larval tiger salamanders (Ambystoma tigrinum). While this preparation was originally developed for recordings with sharp microelectrodes5,6, we describe a method for dual whole-cell voltage clamp recordings from photoreceptors and second-order horizontal and bipolar cells in which we manipulate the photoreceptor's membrane potential while simultaneously recording post-synaptic responses in horizontal or bipolar cells. The photoreceptors of the tiger salamander are considerably larger than those of mammalian species, making this an ideal preparation in which to undertake this technically challenging experimental approach. These experiments are described with an eye toward probing the signaling properties of the synaptic ribbon - a specialized synaptic structure found in a only a handful of neurons, including rod and cone photoreceptors, that is well suited for maintaining a high rate of tonic neurotransmitter release7,8 - and how it contributes to the unique signaling properties of this first retinal synapse.

We describe the preparation of thin retinal slices from aquatic tiger salamanders (Ambystoma tigrinum) and explain how we use these slices to study synaptic processing in the retina by obtaining dual whole-cell voltage clamp recordings from photoreceptors and second-order horizontal and bipolar cells.

両生類網膜スライス標本における感光体と2次ニューロンからの同時ホールセル録音

One of the central tasks in retinal neuroscience  is to understand the circuitry of retinal neurons and how those connections are  responsible for shaping ...

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 ...

Simultaneous Recording of Electroretinography and Visual Evoked Potentials in Anesthetized Rats

The electroretinogram (ERG) and visual evoked potential (VEP) are commonly used to assess the integrity of the visual pathway. The ERG measures the electrical responses of the retina to light stimulation, while the VEP measures the corresponding functional integrity of the visual pathways from the retina to the primary visual cortex following the same light event. The ERG waveform can be broken down into components that reflect responses from different retinal neuronal and glial cell classes. The early components of the VEP waveform represent the integrity of the optic nerve and higher cortical centers. These recordings can be conducted in isolation or together, depending on the application. The methodology described in this paper allows simultaneous assessment of retinal and cortical visual evoked electrophysiology from both eyes and both hemispheres. This is a useful way to more comprehensively assess retinal function and the upstream effects that changes in retinal function can have on visual evoked cortical function.

This protocol describes simultaneous measurement of electroretinogram and visual evoked potentials in anesthetized rats.

麻酔ラットにおける電図および視覚誘発電位の同時記録

The electroretinogram (ERG) and visual evoked potential (VEP) are commonly used to assess the integrity of the visual pathway. The ERG measures the ...

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 ...

Acute In Vivo Electrophysiological Recordings of Local Field Potentials and Multi-unit Activity from the Hyperdirect Pathway in Anesthetized Rats

Converging evidence shows that many neuropsychiatric diseases should be understood as disorders of large-scale neuronal networks. To better understand the pathophysiological basis of these diseases, it is necessary to precisely characterize in which way the processing of information is disturbed between the different neuronal parts of the circuit. Using extracellular in vivo electrophysiological recordings, it is possible to accurately delineate neuronal activity within a neuronal network. The application of this method has several advantages over alternative techniques, e.g., functional magnetic resonance imaging and calcium imaging, as it allows a unique temporal and spatial resolution and does not rely on genetically engineered organisms. However, the use of extracellular in vivo recordings is limited since it is an invasive technique that cannot be universally applied. In this article, a simple and easy to use method is presented with which it is possible to simultaneously record extracellular potentials such as local field potentials and multiunit activity at multiple sites of a network. It is detailed how a precise targeting of subcortical nuclei can be achieved using a combination of stereotactic surgery and online analysis of multi-unit recordings. Thus, it is demonstrated, how a complete network such as the hyperdirect cortico-basal ganglia loop can be studied in anesthetized animals in vivo.

Acute In Vivo Electrophysiological Recordings of Local Field Potentials and Multi-unit Activity from the Hyperdirect Pathway in Anesthetized Rats

In this study, the methodology is presented on how to perform multi-site in vivo electrophysiological recordings from the hyperdirect pathway under urethane anesthesia.

急性<em&gt;インビボ</em&gt;麻酔ラットにおけるHyperdirect経路からの局所場ポテンシャルとマルチユニット活性の電気生理学的記録

Converging evidence shows that many neuropsychiatric diseases should be understood as disorders of large-scale neuronal networks. To better understand the ...

Noninvasive EEG Recordings from Freely Moving Piglets

The method allows the recording of high-quality electroencephalograms (EEGs) from freely moving piglets directly in the pigpen. We use a one-channel telemetric electroencephalography system in combination with standard self-adhesive hydrogel electrodes. The piglets are calmed down without the use of sedatives. After their release into the pigpen, the piglets behave normally&#8212;they drink and sleep in the same cycle as their siblings. Their sleep phases are used for the EEG recordings.

Here, we present a protocol to record telemetric electroencephalograms (EEGs) from freely moving piglets directly in the pigpen without the use of a sedative, making it possible to record typical EEG patterns during non-REM sleep, like spindle bursts.

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 ...

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 ...

Generation of Neurospheres from Mixed Primary Hippocampal and Cortical Neurons Isolated from E14-E16 Sprague Dawley Rat Embryo

Primary neuron culture is an essential technique in the field of neuroscience. To gain deeper mechanistic insights into the brain, it is essential to have a robust in vitro model that can be exploited for various neurobiology studies. Though primary neuron cultures (i.e., long-term hippocampal cultures) have provided scientists with models, it does not yet represent the complexity of brain network completely. In the wake of these limitations, a new model has emerged using neurospheres, which bears a closer resemblance to the brain tissue. The present protocol describes the plating of high and low densities of mixed cortical and hippocampal neurons isolated from the embryo of embryonic day 14-16 Sprague Dawley rats. This allows for the generation of neurospheres and long-term primary neuron culture as two independent platforms to conduct further studies. This process is extremely simple and cost-effective, as it minimizes several steps and reagents previously deemed essential for neuron culture. This is a robust protocol with minimal requirements that can be performed with achievable results and further used for a diversity of studies related to neuroscience.

Presented here is a protocol for the spontaneous generation of neurospheres enriched in neural progenitor cells from high density plated neurons. During the same experiment, when neurons are plated at a lower density, the protocol also results in prolonged primary rat neuron cultures.

E14-E16スプラーグドーリーラット胚から分離された混合原発海馬および皮質ニューロンからの神経球の生成

Primary neuron culture is an essential technique in the field of neuroscience. To gain deeper mechanistic insights into the brain, it is essential to have ...