この研究は、ナイトキャンパス学部奨学生プログラムとフィルアンドペニーナイト財団の寛大な資金提供によって部分的に可能になりました。

ここで詳述されているすべての方法は、IRBプロトコルおよび承認(オレゴン大学IRBプロトコル番号10182017.017)に準拠して実行されました。インフォームドコンセントはすべての被験者から得られた。
1. 到達装置
<ol><li>大きなグラフィックタブレットをデスクトップに平らに置きます。</li>
	<li>調整可能な80-20アルミニウムフレームを使用して、画面を上に向けて、タスクモニター6-8をタブレットの上に平行に配置します(青写真については、ここを確認してください:https://github.com/greenhouselab/Reach_TMS および補足図1)。 注意: この設定により、参加者はタブレット全体に手を伸ばし、到達する腕の視界を遮りながら、タスクモニターに表示されるターゲットを取得できます。</li>
	<li>Kim et al.3 で説明されているセットアップを参考にしてください。</li>
</ol>2. 機械インターフェース
<ol><li>USBポート を介して タブレットをコンピューターに接続します。HDMIポート を介して タスクモニターをコンピューターに接続します。DB-9 ケーブル を介して 背面 TMS ポートをコンピュータに接続します。</li>
	<li>PCI-6220カードDAQ を介して EMGシステムをコンピュータに接続します。フォトダイオードをBNCケーブル を介して EMGシステムに接続します。</li>
</ol>3. フォトダイオードセンサ
<ol><li>BNCケーブルにフォトダイオードセンサーを接続します。フォトダイオードセンサーをテープでタスクモニターの右上隅に固定し、センサーを画面に向けて1cm離≤ようにします。 注意: これにより、タスクモニターに表示される刺激のタイミングが、独立した入力チャネルのアナログデータとして記録されます。</li>
</ol>4. ソフトウェア
<ol><li>MATLAB2018 用のVETAツールボックス26(https://github.com/greenhouselab/Veta)をダウンロードして、データ収集用のハードウェアとインターフェースします。</li>
	<li>実験パラメータの制御とタブレットとのインターフェースのために開発された到達タスクコード(https://github.com/greenhouselab/Reach_TMS)をダウンロードしてください。</li>
</ol>5.参加者のスクリーニングとインフォームドコンセント
<ol><li>TMSに対する禁忌について被験者をスクリーニングする。除外基準には、発作、頭痛、脳外傷、失神、慢性ストレスまたは不安、睡眠の問題、および神経活性薬の個人的または家族歴が含まれます。追加の除外基準には、脳または頭蓋骨の金属インプラント、およびテスト前の24時間でのレクリエーショナルドラッグまたはアルコールの使用が含まれます。選択基準には、右利きと18歳から35歳までの年齢が含まれていました。</li>
	<li>手順と関連するリスクについて書面で説明し、参加者が持つ可能性のあるその他の質問を明確にします。</li>
	<li>参加者からインフォームドコンセントを取得します。</li>
</ol>6.件名の設定
<ol><li>タブレットに面した快適な椅子に被験者を配置します。脚を机の下に置いた状態で、膝が90°に曲がっていることを確認します。</li>
	<li>皮膚を準備し、EMG電極を配置します。<ol><li>きめの細かいサンドペーパーを使用して、右第一背側骨間筋(FDI)、橈骨伸筋、前三角筋の部位、および首の付け根のC4隆起の皮膚をやさしく摩耗させ、TMSパルスによって生成される電気的アーチファクトを検出します。 注意: 筋肉の記録サイトは、ユーザーのニーズに基づいて適応させることができます。</li>
		<li>研磨された各領域を、電極部位ごとに1回アルコール準備パッドで拭き取り、洗浄します。</li>
		<li>各サイトに1つのEMG電極を配置します。電極が筋線維に対して垂直に走っていることを確認してください。接地電極を右肘の骨の隆起に配置します。</li>
		<li>各電極を医療用テープで固定します。</li>
	</ol></li>
	<li>EMG記録の品質を確認してください。VETAツールボックスを使用して、すべてのEMGトレースを視覚化し、アーティファクトがないことを確認します。EMGトレースにノイズが多い場合は、地面が適切に配置され、すべての電極が皮膚に適切に接触していることを確認してください。</li>
</ol>7.経頭蓋磁気刺激
<ol><li>TMS マシンの電源を入れます。</li>
	<li>左M1の刺激 を介して 右FDI筋肉のTMSホットスポットを見つけます。<ol><li>コイルを頭の頂点に対して~5 cm横、2 cm前方に配置し、正中線から~45°離します。</li>
		<li>TMSパルスを4秒ごとに投与し、コイルを前後および内側外側の平面で約5mm刻みで再配置します。</li>
		<li>最大刺激装置出力30%から始めて、MEPが観察されるまでTMS強度を2%刻みで徐々に増加させます。</li>
		<li>可能な限り低い刺激器強度で大部分のパルス(~75%)でMEPを確実に誘発できる最適な位置が特定されたら、10パルスのうち5パルスでピークtoピーク振幅>50μVのMEPを生成する強度レベルを見つけることにより、安静時運動閾値(RMT)を決定します。</li>
		<li>コイルの周囲に沿って参加者の頭に反射テープの薄いストリップをそっと置き、位置をマークします。コイルを手動で保持するか、スタンドを使用してコイルを支えて、コイルの位置を維持します。</li>
	</ol></li>
</ol>8. タスク設定の到達
<ol><li>参加者の右手にベルクログローブをはめて、リラックスしたパワーグリップの姿勢を促進します。</li>
	<li>スタイラスを手袋に取り付け、手を伸ばす動きの間に手をリラックスさせておくように被験者にアドバイスします。</li>
	<li>次のようなタスクの指示を伝えます。 カーソルを画面下部のホーム位置に誘導します。2 つのターゲット位置のいずれかにキューが表示されます。ターゲットが色で塗りつぶされたら、できるだけ速く正確にターゲットに到達します。その後、ホームポジションに戻ります。ホームポジション、キュー、およびターゲットの位置を示します(図1A)。</li>
	<li>スタイラスでターゲットをできるだけ迅速かつ正確にスライスするように参加者を指導します。タスクルームの照明を消して、参加者の腕の動きの視界を覆い隠し、タスクモニターの視認性を向上させます。</li>
</ol>9.タスク設計
<ol><li>Matlab 2018のPsychtoolbox 3.0で視覚刺激の提示を制御します(補足コーディングファイル1)。</li>
	<li>次のパラメーターを使用して、現在のデータを照合します: 20 模擬試験;270回の試験試験。テストトライアルの4/5のTMS。TMSは、準備キューの開始(ベースラインTMS)と一致するか、命令キューの100ミリ秒前(TMSを遅らせる)を同じ頻度で一致させます。全試行の1/10はキャッチトライアルであり、命令的な手がかりは現れません。ホームポジションは、ワークスペースの下部中央に配置された半径2cmの円です。半径1cmの2つの円形ターゲットは、ホームポジションから15cm、正中線から+45°、-45°離れた場所に配置されます。</li>
	<li>イベントの順序と期間を次のように設定します: 準備キューを 900 ミリ秒、命令キューを 900 ミリ秒に設定します。</li>
</ol>10. TMSの管理
<ol><li>VETAツールボックスは、TMSを同時に管理し、EMG https://github.com/greenhouselab/Veta を記録します。</li>
	<li>VETAツールボックスを使用してTMSパルスのタイミングを制御し、選択した行動イベント(つまり、準備キューの開始またはターゲット発症の100ミリ秒前)と一致します。</li>
	<li>分析に十分な数の MEP を確保するために、十分な頻度で TMS を提供します。 注:書かれているように、タスクコードは、ベースラインMEPを引き出すための準備キューの開始時、または遅延MEPを引き出すための命令的キューの100ミリ秒前のいずれかで、合計試行の4/5でTMSパルスを配信します。パラメータは、ユーザーのニーズに応じてコード内で調整できます。TMSのない試験は、TMSがない場合の行動パフォーマンスを評価するために使用できます。これは、TMS がパフォーマンスに与える影響を判断するのに役立ちます。</li>
</ol>

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R., Mashni, A., Michon, K. J., Lavis, H., Vesia, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Measuring+and+manipulating+functionally+specific+neural+pathways+in+the+human+motor+system+with+transcranial+magnetic+stimulation.">Measuring and manipulating functionally specific neural pathways in the human motor system with transcranial magnetic stimulation.</a> Journal of Visualized Experiments. (156), e60706 (2020).</li></ol>

すべての著者は、利益相反がないことを宣言します

上記で概説した方法は、到達行動の文脈で運動準備を研究するための新しいアプローチを提供します。リーチは、運動制御と学習の研究において一般的なモデルタスクを表していますが、リーチ行動に関連するCSダイナミクスを正確に評価する必要があります。TMSは、到達中の離散的な時点でCS活性をキャプチャする非侵襲的で時間的に正確な方法を提供します。ここで説明する?...

目標指向の到達は、人間が外部環境と相互作用し、操作することを可能にする基本的な運動行動です。運動生理学、心理学、神経科学の分野での到達の研究は、さまざまな方法論を含む豊富で広範な文献を生み出しました。到達に関する初期の研究では、ヒト以外の霊長類の直接神経記録を使用して、単一ニューロンレベルでの神経活動を調査しました1,2。より最近の研究では、感覚運動適応を採用して運動学習と制御の性質を探求する行動パラダイムを使用して到達することを調査しています3,4,5。機能的磁気共鳴画像法および脳波検査と組み合わされたこのような行動課題は、ヒトにおける到達中の全脳活動を測定することができる6,7。他の研究では、リーチの準備と実行のさまざまな機能を調査するためにオンラインTMSを適用しています8、9、10、11、12、13、14。ただし、リーチの行動評価とTMSを組み合わせたオープンソースで柔軟なアプローチの必要性が残っています。TMSと行動プロトコルを組み合わせることの有用性は非常によく確立されていますが15、ここでは、オープンソースアプローチを使用して到達するという文脈でのTMSの適用を具体的に検討します。これは、この方法の組み合わせを使用して公開した他のグループがツールをすぐに利用できるようにせず、直接複製を禁止しているという点で斬新です。このオープンソースアプローチにより、複製、データ共有、およびマルチサイト調査の可能性が容易になります。さらに、他の人が同様のツールを使用して新しい研究の質問を追求したい場合は、オープンソースコードはすぐに適応できるため、イノベーションの出発点として機能することができます。
TMSは、正確に制御された時点16で運動システムをプローブする非侵襲的な手段を提供します。一次運動皮質(M1)に適用すると、TMSは標的筋肉の筋電図で測定可能なたわみを誘発することができる。運動誘発電位(MEP)として知られるこの電圧波の振幅は、皮質脊髄(CS)経路の瞬間的な興奮性状態の指標を提供し、CS経路に対するすべての興奮性および抑制性の影響の結果として生じる類似体17である。TMSは、固有のCS興奮性の被験者内測定を確実に提供することに加えて、他の行動学的または運動学的メトリックと組み合わせて、CS活性と行動との関係を時間的に正確な方法で調査することができる。多くの研究は、特にこの方法の組み合わせが膨大な数の行動条件下でMEPを調査することを可能にするため、運動系に関するさまざまな質問に対処するためにTMSと筋電図(EMG)の組み合わせを利用してきました15。これが特に有用であることが証明されている分野の1つは、行動準備の研究であり、ほとんどの場合、単一関節運動の研究を通じてです18。しかし、リーチングなどの自然主義的な多関節運動に関するTMSの研究は比較的少ない。
現在の目標は、行動運動学、オンラインシングルパルスTMS投与、および複数の筋肉からの同時EMG記録を含む遅延応答到達タスクを設計することでした。このタスクには、視覚フィードバックが到達軌道と一致するように、水平方向のモニターを使用したオンライン視覚フィードバックを備えた2次元のポイントツーポイント到達パラダイムが含まれます(つまり、検証フィードバック中の1:1の関係であり、視覚フィードバックとモーションの間の変換はありません)。現在の設計には、視覚運動摂動を伴う一連の試行も含まれています。提供されている例では、これはカーソルフィードバックの20°回転シフトです。以前の研究では、感覚運動適応に関連するメカニズムと計算に関する質問に対処するために、同様の到達パラダイムを使用してきました19、20、21、22、23、24、25。さらに、このアプローチにより、オンライン運動学習中の正確な時点での運動システムの興奮性ダイナミクスを評価することができます。
リーチは学習/適応を調査するための実りある行動であることが証明されているため、この行動の文脈でCS興奮性を評価することは、これらの行動に関与する神経基質に光を当てる大きな可能性を秘めています。これらには、非ヒト霊長類研究で確立されているように、局所的な抑制的影響、同調特性の変化、神経事象のタイミングなどが含まれる場合があります。ただし、これらの特徴は、ヒトおよび臨床集団で定量化することがより困難でした。神経ダイナミクスは、TMSとEMGを組み合わせたアプローチを使用して、ヒトに明白な動きがない場合(すなわち、運動の準備中または安静時)に調査することもできます。
提示されたツールはオープンソースであり、コードは簡単に適応できます。この新しいパラダイムは、到達運動の準備、実行、終了、および適応に関与するメカニズムに関する重要な洞察を生み出します。さらに、この方法の組み合わせは、電気生理学とヒトの到達行動との関係を明らかにする可能性を秘めています。

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>2-Port Native PCI Express&#160;</td>
			<td>StarTech.com</td>
			<td>RS232 Card with 16950 UART&#160;</td>
			<td>Must be compatible with desktop computer</td>
		</tr>
		<tr>
			<td>Adjustable 80-20 aluminum frame</td>
			<td>any</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Alcohol prep pads</td>
			<td>any</td>
			<td></td>
			<td>EMG preparation</td>
		</tr>
		<tr>
			<td>Bagnoli Bipolar Electrodes</td>
			<td>Delsys</td>
			<td>DE 2.1</td>
			<td></td>
		</tr>
		<tr>
			<td>Bagnoli Reference Electrode</td>
			<td>Delsys</td>
			<td>USX2000</td>
			<td>2&#8221; (5cm) Round</td>
		</tr>
		<tr>
			<td>Bagnoli-8 EMG System</td>
			<td>Delsys</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Chair</td>
			<td>any</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Computer monitor for EMG/TMS</td>
			<td>n/a</td>
			<td></td>
			<td></td>
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		<tr>
			<td>Desk</td>
			<td>any</td>
			<td></td>
			<td></td>
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		<tr>
			<td>Desktop Computer</td>
			<td>Dell</td>
			<td>xps 8930</td>
			<td>RAM: 16 GB, Storage: 1TB, Graphics: 1060 6GB&#160;</td>
		</tr>
		<tr>
			<td>EMG electrodes</td>
			<td>Delsys</td>
			<td>Sensor Adhesive Interface</td>
			<td></td>
		</tr>
		<tr>
			<td>Fine grain sandpaper</td>
			<td>any</td>
			<td></td>
			<td>EMG preparation</td>
		</tr>
		<tr>
			<td>Graphics tablet</td>
			<td>Wacom</td>
			<td>Intuos-4 XL</td>
			<td></td>
		</tr>
		<tr>
			<td>Handle of paint roller</td>
			<td>any</td>
			<td></td>
			<td>to be used as stylus handle, hollowed out center must be large enough for stylus to sit securely inside&#160;</td>
		</tr>
		<tr>
			<td>Medical tape</td>
			<td>any</td>
			<td></td>
			<td>To secure EMG electrodes</td>
		</tr>
		<tr>
			<td>PCI-6220 card DAQ</td>
			<td>National Instruments</td>
			<td></td>
			<td>To interface EMG system</td>
		</tr>
		<tr>
			<td>Photodiode Sensor</td>
			<td>Vishay</td>
			<td>BPW21R</td>
			<td>To record timing of task events into EMG trace.</td>
		</tr>
		<tr>
			<td>Rear TMS port</td>
			<td>Magstim</td>
			<td></td>
			<td>Included with TMS machine</td>
		</tr>
		<tr>
			<td>Right-handed polyethylene glove</td>
			<td>any</td>
			<td></td>
			<td>Cut out thumb and index finger of glove to expose FDI muscle</td>
		</tr>
		<tr>
			<td>Sensory Adhesive Interface, 2-slot</td>
			<td>Delsys</td>
			<td>SC-F01</td>
			<td></td>
		</tr>
		<tr>
			<td>Stylus</td>
			<td>Wacom</td>
			<td>Intuos-4 grip pen</td>
			<td></td>
		</tr>
		<tr>
			<td>Tablet-to-Computer USB cable&#160;</td>
			<td>any</td>
			<td></td>
			<td>Included in Tablet purchase</td>
		</tr>
		<tr>
			<td>Task Monitor</td>
			<td>Asus</td>
			<td>VG248</td>
			<td></td>
		</tr>
		<tr>
			<td>TMS coil</td>
			<td>Magstim</td>
			<td>D70 Remote Coil</td>
			<td>7cm diameter, figure-of-eight coil</td>
		</tr>
		<tr>
			<td>TMS machine</td>
			<td>Magstim</td>
			<td>200-2</td>
			<td></td>
		</tr>
		<tr>
			<td>TMS-to-Computer DB9 cable</td>
			<td>any</td>
			<td></td>
			<td>Connects to PCIe Serial Card</td>
		</tr>
		<tr>
			<td>Velcro</td>
			<td>any</td>
			<td></td>
			<td>To be placed on glove and stylus handle</td>
		</tr>
	</tbody>
</table>

assessing corticospinal excitability during goal-directed reaching behavior

リーチは、運動生理学および神経科学の研究で広く研究されている行動です。リーチはさまざまな行動操作を使用して検討されてきましたが、リーチの計画、実行、および制御に関与する神経プロセスの理解には大きなギャップが残っています。ここで説明する新しいアプローチは、2次元の到達タスクと経頭蓋磁気刺激(TMS)および複数の筋肉からの同時筋電図(EMG)記録を組み合わせたものです。この方法は、到達運動の展開中の正確な時点での皮質脊髄活動の非侵襲的検出を可能にする。タスクコード例には、正中線から45°±表示された2つの可能なターゲットを含む遅延応答到達タスクが含まれています。シングルパルスTMSは、準備キューの開始時(ベースライン)または命令的キュー(遅延)の100ミリ秒前のいずれかで、ほとんどのタスク試行で配信されます。このサンプルデザインは、リーチ準備中の皮質脊髄興奮性の変化を調査するのに適しています。サンプルコードには、リーチ準備中の皮質脊髄興奮性に対する適応の影響を調査するための視覚運動摂動(つまり、カーソル±20°回転)も含まれています。タスクパラメータとTMS配信は、到達行動中の運動システムの状態に関する特定の仮説に対処するように調整できます。最初の実施では、TMS試験の83%で運動誘発電位(MEP)が正常に誘発され、すべての試験でリーチ軌道が記録されました。

記載された方法の正常な実行には、錠剤データの記録、EMGトレース、およびMEPの信頼性の高い誘発が含まれる。試験の4/5(216試験)でTMSが実施された270の試験を含む実験が完了した。
データは、25歳から10歳までの16人の参加者(女性8人、男性8人)から収集±、全員が右利きであると自己申告しました。視覚的摂動が行動パフォーマンスに及ぼす有効性を,代表参加者1名の学習?...

Watch this Scientific Journal Video about Assessing Corticospinal Excitability During Goal-Directed Reaching Behavior at JoVE.com

Assessing Corticospinal Excitability During Goal-Directed Reaching Behavior

リーチは、人間が環境と相互作用することを可能にする基本的なスキルです。いくつかの研究は、さまざまな方法論を使用して到達行動を特徴付けることを目的としています。この論文は、タスクパフォーマンスに到達する間の人間の皮質脊髄興奮性の状態を評価するための経頭蓋磁気刺激のオープンソースアプリケーションを提供します。

目標指向到達行動中の皮質脊髄興奮性の評価

Reaching is a widely studied behavior in motor physiology and neuroscience research. While reaching has been examined using a variety of behavioral ...

assessing-corticospinal-excitability-during-goal-directed-reaching

Research

JoVE Journal

Neuroscience

1.6K ビュー.  University of Oregon. リーチは、人間が環境と相互作用することを可能にする基本的なスキルです。いくつかの研究は、さまざまな方法論を使用して到達行動を特徴付けることを目的としています。この論文は、タスクパフォーマンスに到達する間の人間の皮質脊髄興奮性の状態を評価するための経頭蓋磁気刺激のオープンソースアプリケーションを提供します。

ビデオ: Assessing Corticospinal Excitability During Goal-Directed Reaching Behavior

Single Sensillum Recordings in the Insects Drosophila melanogaster and Anopheles gambiae

The sense of smell is essential for insects to find foods, mates, predators, and oviposition sites3. Insect olfactory sensory neurons (OSNs) are enclosed in sensory hairs called sensilla, which cover the surface of olfactory organs. The surface of each sensillum is covered with tiny pores, through which odorants pass and dissolve in a fluid called sensillum lymph, which bathes the sensory dendrites of the OSNs housed in a given sensillum. The OSN dendrites express odorant receptor (OR) proteins, which in insects function as odor-gated ion channels4, 5. The interaction of odorants with ORs either increases or decreases the basal firing rate of the OSN. This neuronal activity in the form of action potentials embodies the first representation of the quality, intensity, and temporal characteristics of the odorant6, 7.
Given the easy access to these sensory hairs, it is possible to perform extracellular recordings from single OSNs by introducing a recording electrode into the sensillum lymph, while the reference electrode is placed in the lymph of the eye or body of the insect. In Drosophila, sensilla house between one and four OSNs, but each OSN typically displays a characteristic spike amplitude. Spike sorting techniques make it possible to assign spiking responses to individual OSNs. This single sensillum recording (SSR) technique monitors the difference in potential between the sensillum lymph and the reference electrode as electrical spikes that are generated by the receptor activity on OSNs1, 2, 8. Changes in the number of spikes in response to the odorant represent the cellular basis of odor coding in insects. Here, we describe the preparation method currently used in our lab to perform SSR on Drosophila melanogaster and Anopheles gambiae, and show representative traces induced by the odorants in a sensillum-specific manner.

Single Sensillum Recordings in the Insects Drosophila melanogaster and Anopheles gambiae

Electrophysiological responses of olfactory sensory neurons to odorants can be measured in insects using single sensillum recordings. In this video article we will demonstrate how to perform single sensillum recordings in the antennae of the vinegar fly (Drosophila melanogaster) and the maxillary palps of the malaria mosquito (Anopheles gambiae).

昆虫における単一感覚子録音キイロショウジョウバエとハマダラカ</em

The sense of smell is essential for insects to find foods, mates, predators, and oviposition sites3. Insect olfactory sensory neurons (OSNs) are enclosed ...

神経科学

T-maze Forced Alternation and Left-right Discrimination Tasks for Assessing Working and Reference Memory in Mice

Forced alternation and left-right discrimination tasks using the T-maze have been widely used to assess working and reference memory, respectively, in rodents. In our laboratory, we evaluated the two types of memory in more than 30 strains of genetically engineered mice using the automated version of this apparatus. Here, we present the modified T-maze apparatus operated by a computer with a video-tracking system and our protocols in a movie format. The T-maze apparatus consists of runways partitioned off by sliding doors that can automatically open downward, each with a start box, a T-shaped alley, two boxes with automatic pellet dispensers at one side of the box, and two L-shaped alleys. Each L-shaped alley is connected to the start box so that mice can return to the start box, which excludes the effects of experimenter handling on mouse behavior. This apparatus also has an advantage that in vivo microdialysis, in vivo electrophysiology, and optogenetics techniques can be performed during T-maze performance because the doors are designed to go down into the floor. In this movie article, we describe T-maze tasks using the automated apparatus and the T-maze performance of &alpha;-CaMKII+/- mice, which are reported to show working memory deficits in the eight-arm radial maze task. Our data indicated that &alpha;-CaMKII+/- mice showed a working memory deficit, but no impairment of reference memory, and are consistent with previous findings using the eight-arm radial maze task, which supports the validity of our protocol. In addition, our data indicate that mutants tended to exhibit reversal learning deficits, suggesting that &alpha;-CaMKII deficiency causes reduced behavioral flexibility. Thus, the T-maze test using the modified automatic apparatus is useful for assessing working and reference memory and behavioral flexibility in mice.

This article presents the protocol of T-maze tests using a modified automated apparatus for assessing the learning and memory functions in mice.

マウスで作業していて、メモリの参照を評価するためのT-迷路強制交替と左右弁別課題

Forced alternation and left-right discrimination tasks using the T-maze have been widely used to assess working and reference memory, respectively, in ...

Nerve Excitability Assessment in Chemotherapy-induced Neurotoxicity

Chemotherapy-induced neurotoxicity is a serious consequence of cancer treatment, which occurs with some of the most commonly used chemotherapies1,2. Chemotherapy-induced peripheral neuropathy produces symptoms of numbness and paraesthesia in the limbs and may progress to difficulties with fine motor skills and walking, leading to functional impairment. In addition to producing troubling symptoms, chemotherapy-induced neuropathy may limit treatment success leading to dose reduction or early cessation of treatment. Neuropathic symptoms may persist long-term, leaving permanent nerve damage in patients with an otherwise good prognosis3. As chemotherapy is utilised more often as a preventative measure, and survival rates increase, the importance of long-lasting and significant neurotoxicity will increase.
There are no established neuroprotective or treatment options and a lack of sensitive assessment methods. Appropriate assessment of neurotoxicity will be critical as a prognostic factor and as suitable endpoints for future trials of neuroprotective agents. Current methods to assess the severity of chemotherapy-induced neuropathy utilise clinician-based grading scales which have been demonstrated to lack sensitivity to change and inter-observer objectivity4. Conventional nerve conduction studies provide information about compound action potential amplitude and conduction velocity, which are relatively non-specific measures and do not provide insight into ion channel function or resting membrane potential. Accordingly, prior studies have demonstrated that conventional nerve conduction studies are not sensitive to early change in chemotherapy-induced neurotoxicity4-6. In comparison, nerve excitability studies utilize threshold tracking techniques which have been developed to enable assessment of ion channels, pumps and exchangers in vivo in large myelinated human axons7-9.
Nerve excitability techniques have been established as a tool to examine the development and severity of chemotherapy-induced neurotoxicity10-13. Comprising a number of excitability parameters, nerve excitability studies can be used to assess acute neurotoxicity arising immediately following infusion and the development of chronic, cumulative neurotoxicity. Nerve excitability techniques are feasible in the clinical setting, with each test requiring only 5 -10 minutes to complete. Nerve excitability equipment is readily commercially available, and a portable system has been devised so that patients can be tested in situ in the infusion centre setting. In addition, these techniques can be adapted for use in multiple chemotherapies.
In patients treated with the chemotherapy oxaliplatin, primarily utilised for colorectal cancer, nerve excitability techniques provide a method to identify patients at-risk for neurotoxicity prior to the onset of chronic neuropathy. Nerve excitability studies have revealed the development of an acute Na+ channelopathy in motor and sensory axons10-13. Importantly, patients who demonstrated changes in excitability in early treatment were subsequently more likely to develop moderate to severe neurotoxicity11. However, across treatment, striking longitudinal changes were identified only in sensory axons which were able to predict clinical neurological outcome in 80% of patients10. These changes demonstrated a different pattern to those seen acutely following oxaliplatin infusion, and most likely reflect the development of significant axonal damage and membrane potential change in sensory nerves which develops longitudinally during oxaliplatin treatment10. Significant abnormalities developed during early treatment, prior to any reduction in conventional measures of nerve function, suggesting that excitability parameters may provide a sensitive biomarker.

This abstract describes a novel method to assess the development of neurotoxicity in patients receiving chemotherapy treatment. While conventional assessment methods are limited in their ability to detect early changes in nerve function, nerve excitability techniques provide early identification of patients at risk of severe neurotoxicity and insight into pathophysiology.

化学療法誘発性神経の神経興奮性の評価

Chemotherapy-induced neurotoxicity is a serious consequence of cancer treatment, which occurs with some of the most commonly used chemotherapies1,2. ...

Unilateral Pyramidotomy of the Corticospinal Tract in Rats for Assessment of Neuroplasticity-inducing Therapies

The corticospinal tract (CST) can be completely severed unilaterally in the medullary pyramids of the rodent brainstem. The CST is a motor tract that has great importance for distal muscle control in humans and, to a lesser extent, in rodents. A unilateral cut of one pyramid results in loss of CST innervation of the spinal cord mainly on the contralateral side of the spinal cord leading to transient motor disability in the forelimbs and sustained loss of dexterity. Ipsilateral projections of the corticospinal tract are minor. We have refined our surgical method to increase the chances of lesion completeness. We describe postsurgical care. Deficits on the Montoya staircase pellet reaching test and the horizontal ladder test shown here are detected up to 8 weeks postinjury. Deficits on the cylinder rearing test are only detected transiently. Therefore, the cylinder test may only be suitable for detection of short term recovery. We show how, electrophysiologically and anatomically, one may assess lesions and plastic changes. We also describe how to analyse fibers from the uninjured CST sprouting across the midline into the deprived areas. It is challenging to obtain &#62;90% complete lesions consistently due to the proximity to the basilar artery in the medulla oblongata and survival rates can be low. Alternative surgical approaches and behavioural testing are described in this protocol. The pyramidotomy model is a good tool for assessing neuroplasticity-inducing treatments, which increase sprouting of intact fibers after injury.

The corticospinal tract, one of the major sensorimotor tracts, can be lesioned unilaterally in the rodent brainstem in order to test neuroplasticity-inducing therapies for the central nervous system. This surgical procedure (&#8220;pyramidotomy&#8221;) and postoperative assessments are described in this protocol.

神経可塑性を誘導する治療法の評価のためのラットの皮質脊髄路の片側Pyramidotomy

The corticospinal tract (CST) can be completely severed unilaterally in the medullary pyramids of the rodent brainstem. The CST is a motor tract that has ...

A Procedure for Implanting a Spinal Chamber for Longitudinal In Vivo Imaging of the Mouse Spinal Cord

Studies in the mammalian neocortex have enabled unprecedented resolution of cortical structure, activity, and response to neurodegenerative insults by repeated, time-lapse in vivo imaging in live rodents. These studies were made possible by straightforward surgical procedures, which enabled optical access for a prolonged period of time without repeat surgical procedures. In contrast, analogous studies of the spinal cord have been previously limited to only a few imaging sessions, each of which required an invasive surgery. As previously described, we have developed a spinal chamber that enables continuous optical access for upwards of 8 weeks, preserves mechanical stability of the spinal column, is easily stabilized externally during imaging, and requires only a single surgery. Here, the design of the spinal chamber with its associated surgical implements is reviewed and the surgical procedure is demonstrated in detail. Briefly, this video will demonstrate the preparation of the surgical area and mouse for surgery, exposure of the spinal vertebra and appropriate tissue debridement, the delivery of the implant and vertebral clamping, the completion of the chamber, the removal of the delivery system, sealing of the skin, and finally, post-operative care. The procedure for chronic in vivo imaging using nonlinear microscopy will also be demonstrated. Finally, outcomes, limitations, typical variability, and a guide for troubleshooting are discussed.

A Procedure for Implanting a Spinal Chamber for Longitudinal In Vivo Imaging of the Mouse Spinal Cord

In this video, we describe a procedure for implanting a chronic optical imaging chamber over the dorsal spinal cord of a live mouse. The chamber, surgical procedure, and chronic imaging are reviewed and demonstrated.

縦断のために脊髄商工を移植するための手順<em&gt;インビボ</emマウス脊髄の&gt;イメージング

Studies in the mammalian neocortex have enabled unprecedented resolution of cortical structure, activity, and response to neurodegenerative insults by ...

Assays to Detect UV-reflecting Structures and Determine their Importance in Mate Preference using the Sailfin Molly Poecilia latipinna

Many organisms use cues and signals beyond human sensitivity during social interactions. It is important to take into account how organisms perceive their worlds when trying to understand their behavior and ecology. Sensitivity to the ultraviolet spectrum (UV; 300 - 400 nm) is found across multiple genera of birds, fish, reptiles, amphibians, and even mammals. This protocol describes a technique for examining organisms for the presence of UV-reflecting structures and a method for testing whether these cues are used as social signals in the context of mate choice. A spectrophotometer is used to detect the presence of UV reflectance and variation in reflective intensity between individuals and sexes. An example of this technique is presented in which a dichotomous mate choice test exposes sexually receptive individuals to opposite sex individuals whose visual appearance can be manipulated by filters that either transmit full spectrum or block UV wavelengths. This system allowed for the determination that female, but not male, sailfin mollies (Poecilia latipinna) were using UV markings as part of their mating decisions. These types of studies serve to expand our knowledge of the range of organisms that utilize UV and provide insight into how UV plays a role in their lives.

Assays to Detect UV-reflecting Structures and Determine their Importance in Mate Preference using the Sailfin Molly Poecilia latipinna

This protocol outlines the use of spectrophotometry to detect ultraviolet-reflecting structures on organisms (in this example, the sailfin molly Poecilia latipinna) and describes dichotomous choice tests for fish that allow inferences to be made on the role of ultraviolet cues during mate selection.

UV反射構造を検出してのSailFinモリーを使用して、メイト優先でその重要性を決定するためのアッセイ<em&gt; Poecilia latipinna</em

Many organisms use cues and signals beyond human sensitivity during social interactions. It is important to take into account how organisms perceive their ...

Operant Protocols for Assessing the Cost-benefit Analysis During Reinforced Decision Making by Rodents

Reinforcement-guided decision making is the ability to choose between competing courses of action based on the relative value of the benefits and their consequences. This process is integral to the normal human behavior and has been shown to be disrupted by neurological and psychiatric disorders such as addiction, schizophrenia, and depression. Rodents have long been used to uncover the neurobiology of human cognition. To this end, several behavioral tasks have been developed; however, most are non-automated and are labor-intensive. The recent development of the open-source microcontroller has enabled researchers to automate operant-based tasks for assessing a variety of cognitive tasks, standardizing the stimulus presentation, improving the data recording and consequently, improving the research output. Here, we describe an automated delay-based reinforcement-guided decision-making task, using an operant T-maze controlled by custom-written software programs. Using these decision-making tasks, we show the changes in the local field potential activities in the anterior cingulate cortex of a rat whilst it performs a delay-based cost-and-benefit decision-making task.

A cost-benefit analysis is a weighing-scale approach that the brain performs during the course of decision making. Here, we propose a protocol to train rats on an operant-based decision-making paradigm where rats choose higher rewards at the expense of waiting for 15 s to receive them.

齧歯動物による補強意思の中に費用便益分析を評価するためのオペラントのプロトコル

Reinforcement-guided decision making is the ability to choose between competing courses of action based on the relative value of the benefits and their ...

Generation of 3-D Collagen-based Hydrogels to Analyze Axonal Growth and Behavior During Nervous System Development

This protocol uses natural type I collagen to generate three-dimensional (3-D) hydrogel for monitoring and analyzing the axonal growth. The protocol is centered on culturing small pieces of embryonic or early postnatal rodent brains inside a 3-D hydrogel formed by the rat tail tendon-derived type I collagen with specific porosity. Tissue pieces are cultured inside the hydrogel and confronted to specific brain fragments or genetically-modified cell aggregates to produce and secrete molecules suitable for creating a gradient inside the porous matrix. The steps of this protocol are simple and reproducible but include critical steps to be considered carefully during its development. Moreover, the behavior of growing axons can be monitored and analyzed directly using a phase-contrast microscope or mono/multiphoton fluorescence microscope after fixation by immunocytochemical methods.

Here, we provide a method for analyzing the behavior of growing axons in 3D matrices, mimicking their natural development.

神経系発達中の軸の成長と挙動を解析する3Dコラーゲンベースのヒドロゲルの生成

This protocol uses natural type I collagen to generate three-dimensional (3-D) hydrogel for monitoring and analyzing the axonal growth. The protocol is ...

Measuring Neural Mechanisms Underlying Sleep-Dependent Memory Consolidation During Naps in Early Childhood

Sleep is critical for daily functioning. One important function of sleep is the consolidation of memories, a process that makes them stronger and less vulnerable to interference. The neural mechanisms underlying the benefit of sleep for memory can be investigated using polysomnography (PSG). PSG is a combination of physiological recordings including signals from the brain (EEG), eyes (EOG), and muscles (EMG) that are used to classify sleep stages. In this protocol, we describe how PSG can be used in conjunction with behavioral memory assessments, actigraphy, and parent-report to examine sleep-dependent memory consolidation. The focus of this protocol is on early childhood, a period of significance as children transition from biphasic sleep (consisting of a nap and overnight sleep) to monophasic sleep (overnight sleep only). The effects of sleep on memory performance are measured using a visuospatial memory assessment across periods of sleep and wakeful-rest. A combination of actigraphy and parent report is used to assess sleep rhythms (i.e., characterizing children as habitual or non-habitual nappers). Finally, PSG is used to characterize sleep stages and qualities of those stages (such as frequencies and the presence of spindles) during naps. The advantage of using PSG is that it is the only tool currently available to assess sleep quality and sleep architecture, pointing to the relevant brain state that supports memory consolidation. The main limitations of PSG are the length of time it takes to prepare the recording montage and that recordings are typically taken over one sleep bought. These limitations can be overcome by engaging young participants in distracting tasks during application and combining PSG with actigraphy and self/parent-report measures to characterize sleep cycles. Together, this unique combination of methods allows for investigations into how naps support learning in preschool children.

This protocol describes methods used to examine neural mechanisms underlying sleep-dependent memory consolidation during naps in early childhood. It includes procedures for examining the effect of sleep on behavioral memory performance, as well as the application and recording of both polysomnography and actigraphy.

幼児期の昼寝中の睡眠依存性記憶統合の基礎となる神経機構の測定

Sleep is critical for daily functioning. One important function of sleep is the consolidation of memories, a process that makes them stronger and less ...

Determining the Functional Status of the Corticospinal Tract Within One Week of Stroke

High interindividual variability in the recovery of upper limb (UL) function after stroke means it is difficult to predict an individual&#39;s potential for recovery based on clinical assessments alone. The functional integrity of the corticospinal tract is an important prognostic biomarker for recovery of UL function, particularly for those with severe initial UL impairment. This article presents a protocol for evaluating corticospinal tract function within 1 week of stroke. This protocol can be used to select and stratify patients in trials of interventions designed to improve UL motor recovery and outcomes after stroke. The protocol also forms part of the PREP2 algorithm, which predicts UL function for individual patients 3 months poststroke. The algorithm sequentially combines a UL strength assessment, age, transcranial magnetic stimulation, and stroke severity, within a few days of the stroke. The benefits of using PREP2 in clinical practice are described elsewhere. This article focuses on the use of a UL strength assessment and transcranial magnetic stimulation to evaluate corticospinal tract function.

This protocol is for evaluating corticospinal tract function within 1 week of stroke. It can be used to select and stratify patients in trials of interventions designed to improve upper limb motor recovery and outcomes and in clinical practice for predicting upper limb functional outcomes 3 months after stroke.

脳卒中1週間以内のコルチコ脊柱管の機能状態の判定

High interindividual variability in the recovery of upper limb (UL) function after stroke means it is difficult to predict an individual&#39;s potential ...