Kriss Franklin, Amanda Pollard and Justine Shaffer assisted in animal training and data collection. Sarah Freeman and Alisa Ivanova contributed to data analysis. This study is supported by WVU School of Medicine Start-Up, NIH/NIGMS U54GM104942, and NIH CoBRE P20GM109098.

次のトレーニングパラダイムは、平均的な成人のスプラーグドーリーラットの位相調整の分析を採用しています。本明細書に記載されたプロトコルは、あなたの機関の動物のケアのガイドラインに従っていることを確認してください。本研究におけるすべての手順は、医学のウェストバージニア大学での実験動物福祉（OLAW）のための制度的動物実験委員会（IACUC）およびOfficeに基づいて行われた実験の使用のための国立衛生研究所のガイドラインを遵守していました動物。 1.機器のセットアップ<ol><li>アルミニウムとブレースオープントップのプラスチックの箱は155センチメートルX 104センチメートル（ 図1）を測定し 、各コーナーにサポートとして、非対称歩道を構築します。同じ側​​に連続する各ペグは歩幅を定義するように、ボックスの周囲に沿って、別のペグの配置を可能にするために、両側の溝付きアルミバー付きのボックスの上部エッジを引き締めます。 </li&gt; <li>それぞれの側に表さ条件を分離し、各コーナーに20センチメートル×20センチプラットフォーム（合計4）を配置します。この距離は、1匹のラットのステップサイクルによって横断距離を含めるために十分であるべきです。 <ol><li> 20センチメートルX 1センチメートル×0.5センチ寸法のアルミ製のペグを使用してください。ベンドそれぞれの上部には、足配置プラットフォームを生成するために先端から2.5センチメートルペグ。 </li><li>レベルの水平配置を確実にするために、同じ距離で機械加工穴を通ってスライドする内側ブラケットを使用して、溝付きのバーにペグを固定します。ドライバーと定規を使って位置を調整します。平均ラットの足の大きさにほぼ対応1cmのペグ幅を使用します。シンナーや幅広ペグのいずれか不快であるか、足配置の変動を増加させます。 </li></ol></li><li> 3正確なステッピングチャレンジのいずれかの条件を生成するためにそれぞれの側にペグの配置を操作します。 <ol><li>左の私を設定することで、15cmのストライド長（SL15）対称運動タスクを生成しますNTER-歩幅（ リットル ISL）と歩幅（7.5センチ）の半分に右間のストライド長（r個の ISL）。 </li><li> 6.0センチメートルにリットルの ISLおよびR ISLの長さを変更することにより、追加の対称条件（SL12）を課しています。 </li><li>左右にペグの間の距離を変化させることにより、非対称なタスクを生成する、インター歩幅と呼ばれます。非対称モータシステムに挑戦するには、左側（L6R9条件）または右（L9R6）側のいずれか短い間ストライド長を課すことを20％Lの ISL と r の ISLを変更します。 1.5センチメートル摂動を6cm の L ISLおよび9 L6R9条件のCM、または9センチの L ISLおよびL9R6条件で6センチの R ISL の R ISLを課します</li></ol></li><li>ラットでは、好ましい15センチメートルでSL12を除くすべての条件に歩幅を保ちます。 </li><li>便宜上、歩道の長辺を左またはrのどちらかに有利な非対称の条件を割り当てます対象のIGHT側、対称制御条件のための2つの短辺を確保しています。 </li><li>ようにセットアップは、少なくとも60ヘルツのサンプリングレートでの高精細カメラペグの手足の配置は、カメラは約7手順をカバーする視野と歩道に垂直に指しで遮られていません。プラットフォームに近接した最初と最後のステップは無視されます。 </li></ol>装置2.トレーニング<ol><li> 例えば 、標準的なトレーニングリソースを使用してください。基本BiomethodologyでNIHのトレーニング実験用ラットのために、げっ歯類の一般的な行動訓練を理解します。 </li><li>トレーニングの初めに、少なくとも5分間、20×20センチのプラットフォーム上に配置し、報いることにより、被験者を順応させます。その後、食品の報酬を提示することによって、次のプラットフォームに1センチメートル間歩幅でペグ配置全体の動物を導きます。口頭およびプラットフォームに到達するためのふれあいを動物に報います。 </li><li> AFTER 5のトレーニングを実行すると、スペースの余分な1ペグ - 2センチメートル離れて、次の5トレーニングランを行います。 （ - 35ステップ20）、本明細書に記載された繰り返し数は、統計学的に適切なサンプルサイズを生成するのに十分です。 <ol><li>動物がよりゆっくりと足踏み（無停止）と姿勢（バックアーチ）の一貫性によって判断されるようにタスクを取得した場合、（長いストライドでトレーニングを再開する前に、短いストライド長（S12）で、これらのスキルの強化に関する訓練を集中最終的に希望の歩幅に近づいS15）。 </li><li>新しい間隔はタスクに不安や不快感を誘発する場合、以前の設定にペグを再調整し、訓練パラダイムを繰り返します。 </li><li>適切なインターストライドの長さが4つの条件と運動基準が満たされているために達成されるまで、このトレーニングを続行します。我々の経験では、ラットは、トライアルを開始するための手がかりとして、ボーカル励ましによく反応します。トレーニングが提供されるテストは、同じ日に行うことができます被験者は、タスクを実行するために動機づけられています。 注：次のように運動の基準は以下の通りです。ウォーキングは一貫しており、停止または失策を必要としません。ヘッドボビングは最小です。バックはアーチ型で、尾が運動中に発生します。各肢は明らかに発症時の歩道の直交ビューから見えると立脚期のオフセットです。本研究は、他の織り付けの動作ではなく、歩くだけに焦点を当てて、この選択プロセスが不可欠です。 </li></ol></li></ol> 3.テストとデータ分析<ol><li>無作為化セッションのデザインを使用して（セクション1.3で説明）、S12、S15、L9R6およびL6R9タスクの試験動物。タスク内の適応を回避するために改行を使用します。 </li><li>少なくとも60ヘルツのサンプリングレートでの高精細カメラで録音セッション。ビデオ編集ソフトウェアに再サンプリングして、さらなる分析のための唯一の歩行発作を選択せず​​にインポートビデオ録画。 </li><li>マークが発症し、運動学的相のオフセット各被験者からのビデオ録画インチ</li><li>ここでは、手動でスタンスの開始時間を特定し、スタンス開始が上の手足の配置に関連したモーションブラーの損失によって示されているフレーム単位で各肢のために相殺するためにMATLABで書かれたカスタムソフトウェアと呼ばれるvideoaを使用四肢リフトオフの開始時に発生するオフセットペグ、及び姿勢は、モーションブラーの最初の証拠によって示されています。 </li><li>二つの連続動スタンスオンセット間の残り時間として遊脚相の持続時間を計算します。歩行は、ダブルスイング相（地面からの両方前肢や後肢）が含まれている場合例えば 、地上四足歩行と一致していない任意の動作を除外します。、進むから分析します。 </li><li>対応するステップのサイクル持続時間の関数として、各フェーズの期間をプロットします。線形回帰モデル（TPHASE = B1 + B2 * TC）との関係はTPHASEが電子である、Tcはサイクル期間で各肢のために取得したキャプチャither テ伸筋関連スタンスまたはTfと 、屈筋関連のスイングで、B1、B2が回帰モデルの実験定数（オフセットとスロープ）です。 注：傾き（B2）は、移動速度の変化に伴う位相持続時間の変化量を表します。 </li><li>非対称性指数（AI）を計算するために、各肢のための式1および2（ 図2C）を使用します。どちらの式はそれらの合計の2つの値の違いを正規化し、単純な比率の同じ形を持っています。 <ol><li>式1を用いて、左スタンス変調（L）と右（R）の手足の傾きの差を使用して水平方向の差（AI H）を算出します。同様に、フロント/前方（a）とバック/後方（P）手足からスロープを使って上下非対称（AIのV）を算出します 。これらの二つの式を適用した結果を&lt;1に対応するXY 4 点 ）前肢の非対称のデータセットでありますem&gt;のAAIの時間; 2）後肢非対称、 パイの H; 3）前肢・後肢非対称性を残し、 ライ V; 4）右前肢・後肢非対称、RAIの V。 </li><li>すべての四肢全体の非対称性を視覚的に表現するためのパッチ（ 図2B）としてこれらの値をプロットします。 </li></ol></li><li>前肢のパラメータとその反対側の後肢（式3、 図2C）との間に斜めの結合を評価するために、対角指数（DI）を計算します。 </li><li>手段分析19の事後比較で一方向ANOVAを用いて統計的有意性について| - |（AIL6R9 AIL9R6ΔAI=）、DIと同様に非対称性を、対向の条件との間に4のAIの違いをテスト</li></ol>

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T., Phillips, C. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Striatal+and+pyramidal+lesions+in+the+cat.">Striatal and pyramidal lesions in the cat.</a> Brain. 69 (4), 264-279 (1946).</li><li>Beloozerova, I. N., Sirota, M. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+Role+of+the+Motor+Cortex+in+the+Control+of+Accuracy+of+Locomotor+Movements+in+the+Cat.">The Role of the Motor Cortex in the Control of Accuracy of Locomotor Movements in the Cat.</a> Journal of Physiology. 461, 1-25 (1993).</li><li>Hill, K. D., Goldie, P. A., Baker, P. A., Greenwood, K. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Retest+reliability+of+the+temporal+and+distance+characteristics+of+hemiplegic+gait+using+a+footswitch+system.">Retest reliability of the temporal and distance characteristics of hemiplegic gait using a footswitch system.</a> Archives of Physical Medicine and Rehabilitation. 75 (5), 577-583 (1994).</li><li>Hillyer, J. E., Joynes, R. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+new+measure+of+hindlimb+stepping+ability+in+neonatally+spinalized+rats.">A new measure of hindlimb stepping ability in neonatally spinalized rats.</a> Behavioural Brain Research. 202 (2), 291-302 (2009).</li></ol>

The authors have nothing to disclose.

The rationale for this study was to develop a behavioral task that quantitatively assesses the changes in precise control of asymmetric locomotor behaviors. The existence of the spinal CPG has been functionally demonstrated for some time20, but the anatomical and functional characteristics that describe its mechanism as well as its modulatory inputs from descending or sensory feedback pathways have not been characterized until the past decade6,21,22. The current consensus is that the intrinsic spinal, sensory feedback, and descending commands are tightly integrated in the generation of locomotor behavior1
The asymmetric precise foot placement task presented herein is designed to functionally challenge the control systems responsible for the dexterous asymmetric control of stepping known to require cortical inputs23,24. This performance was assessed relative to the symmetric tasks that are less reliant on the descending cortical and brainstem control. Thus, we have an opportunity to discern the contributions of the spinal and descending pathways. Since the motor cortex is directly involved in the modulation of muscle phases during locomotion, reaching and postural adjustments9,10,25, the analysis of phase modulation in response to imposed asymmetric precise stepping tasks provides a basis for describing changes in volitional control. This is evident in the significant lateralized phase modulation between left- and right-favored tasks, characterized by the differences in asymmetry indices. We have also observed changes in whole body coordination that required diagonal coupling between contralateral forelimbs and hindlimbs, characterized by differences in the diagonal index.
Both focal stroke26,27 and spinal cord hemilesion28,29 animal models cause mild to moderate movement deficits akin to those observed clinically. In animal models, cortical lesioning of the corticospinal tract impedes or prevents precise stepping30,31. The application of our methodology to the characterization of cortical impairment in stroke models is yet to be described, though some of our preliminary data on rats with middle cerebral artery occlusions showed increased AI, and even a negative slope of the stance phase with increasing cycle duration for the limb on the side contralateral to stroke. This may correspond to a delay in the onset of consecutive locomotor phases, which is consistent with an asymmetry in both the step length ratio and the single limb support time observed in post-stroke patients15,32.
One limitation of this method is that it is inappropriate for the analysis of severely affected animals. However, this subgroup is not necessarily the focus of attention in studies of hemiparetic animals. Furthermore, subjective tracking of this type of deficit requires additional sub scales that may also be associated with high inter-rater variability, creating demand for gross computational methodology33. Thus, the challenge remains not in the assessment of deficits in the severely affected animals, but in the assessment of the mild to severe subgroup. Moreover, the ability to distinguish damage to specific hierarchical areas has been virtually impossible in a non-invasive method. The experimental setup presented herein is an effective tool for the evaluation of moderate impairment by monitoring modulatory activity of the motor antagonistic phases that drive the CPG with different speed demands, presumably contributed by higher order factors of the motor control hierarchy6.

生き残った個体群における脳卒中後の罹患率は、両方のヒトでの定量的評価のための課題を提起する神経学的障害1のストロークと動物モデルを投稿粗大運動機能障害が含まれます。臨床の現場では、これらの運動障害は、大多数の患者が示す厳しいのではなく、中程度の障害に対してより敏感である主観的な基準を用いて測定されます。同様に、動物における損傷後の運動行動のような主観的評価は共通であり、 例えば 、バッソ、ビーティー、とブレスナハン（BBB）運動スケール法2,3。これらの主観的な評価方法は、四足動物モデルおよびヒトにおける歩行リハビリテーション研究間の変換を支援しているが、個別の筋肉群の活性に関連する運動障害の詳細は、評価されていません。また、脳血管障害における運動障害の推定犯人として運動に運動皮質の貢献の評価、彼らはオープンフィールドまたはリニア歩行タスクに依存しているとしてだけで、間接的にも、最も新しい自動化された定量的方法4,5を使用して得ることができます 。これらのタスクは、皮質の貢献を必要とせず、脊髄の神経機構、 すなわち 、神経損傷のほとんどの動物モデルで倹約された中枢パターン発生器（CPG）ネットワーク、 例えば 、spinalized動物6によって行うことができる- 。。8 。これらの脊髄のメカニズムに不可欠な皮質の貢献は、実験的に予想される姿勢調整9および10に到達するだけでなく、正確なステッピング10が必要なタスクに関与しています。 さらに、ほとんどの神経損傷は非対称です。例えば、ストロークは、片麻痺の原因となる、すなわち 、非対称歩行11につながる体の片側の弱さ- 。14。片麻痺歩行の非対称性は非対称spatiotemporによって生成されます最も重要な伸筋関連立脚相の短縮と麻痺側15,16のステップサイクルの屈筋関連遊脚相の延長で明らかにアル筋肉の活性化。この傾向は、まだ健康や麻痺の動物における運動速度の範囲で検討されていません。現在の研究では、各工程におけるサイクル持続時間の関数としてのスイングやスタンス相の持続時間との関係を説明した位相期間特性17の分析を採用しました。得られた線形回帰モデルは、その後、さらに、すべての四肢を横切る非対称性の分析を説明しました。 私たちは、正確なステッピング運動タスクに基づいて、四足動物の運動系に皮質入力を降順の活性を評価するための新規な低コストの方法を報告しています。このタスクは、歩行速度の自然な範囲での足の配置の要求を課すことによって運動野に挑戦するように設計されています。加えて、足配置の要件を優先的運動系の左側または右側に挑戦するために操作されています。同様の運動タスクでは、メッツ＆ウィショー（2009）は、ラットにおいて、不規則なラング歩道上で逃したステップ数を故障率を調べました。私たちの方法は、これまでの研究と相補的であり、それは「成功」に位相制御の品質を詳細に18を繰り返します。

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asymmetric walkway: a novel behavioral assay for studying asymmetric locomotion

行動アッセイは、一般的に中枢神経系（CNS）における感覚運動障害の評価のために使用されます。げっ歯類で運動障害を定量化するための最も洗練された方法は、制約のない歩行地上の微小擾乱を測定することである（ 例えば 、手動のBBBスコアまたは自動キャットウォーク）。しかし、皮質の入力は脊髄中枢パターン発生器（CPG）によって生成される基本的な運動の生成に必要とされません。このように、制約のない歩行タスクは間接的にしかによる運動皮質の障害に運動障害をテストします。本研究では、小説、脊髄CPGに皮質の入力を評価し、正確な足の配置運動タスクを提案します。計装ペグ方が左右で非対称に運動障害を模倣対称および非対称運動タスクを課すために使用されました。私たちは、県と歩行時の前肢スタンス位相特性の20％農産物の変更の等距離間ストライドの長さからずれることを示しています歩幅を誤りました。さらに、我々は非対称の歩道は、皮質の制御信号によって生成された行動の結果の測定を可能にすることを提案します。これらの措置は、皮質損傷後の機能障害の評価に関連しています。

図2は 、単一の代表的な主題のための運動の作業中に非対称性の分析を示します。 （ - 400 gで、 図3 250）の値は、個々に全ての被験者から（ 図2）式1および2を使用して、すべての条件について、図8の雌Sprague-Dawleyラットの複合データから計算しました。一般的に、前肢立脚相の変調は、歩行状態が好まれたために側（短いISL）の優先側（長いISL）上の立脚相がのより大きな部分を占める傾向にあったという概念と一致少ないました歩行減少の速度として好ま手足に比べてステップサイクル。 条件L9R6及びL6R9（ΔAI）から得られた対応する非対称性指数との差はconservativで一方向ANOVA（α= 0.05）、事後t検定を用いて試験しました<html> MATLABでanova1とmultcompare機能を使用して電子ボンフェローニ補正（調整α= 0.0125）。全体的に、群間の差は、た（p = 0.002）に有意でした。前肢間の非対称性に対応する前方水平非対称性指数（Δaを AI h）が左に好ま（L6R9）と右好ま（L9R6）条件（ 図4A）の間（P = 0.006）が有意に異なっていました。右上下非対称指数（ΔRのAI V）の条件の違いは傾向を示したが、それはゼロ（P = 0.031、α= 0.0125）と有意差はなかったです。同様に、2つの非対称の条件（ 図4B）との間に対角指数の大幅な差（p = 0.020、α= 0.05）を発見しました。 ANOVA試験は、異なるタスクでDIの間に有意差は認められなかったが、追加のアルファ補正を必要としない唯一の事後t検定は、ありました。 1 &quot;&gt;このメソッドは、非対称の足の配置を解決するために、動物の自然な能力に依存していたように、いくつかの動物は事後手足がスイングで同時にあったギャロップのような挙動を示すことができる：「キープtogether.withinページ=をFO&quot;トン。この歩行は3匹の動物で観察され、動作がさらなる分析から除外されました。 <img alt="図1" src="/files/ftp_upload/52921/52921fig1.jpg" /> 1.歩道モデル図。歩幅（SL）との関係で（A）対称および非対称歩行タスクに使用歩道の模式図。（B）ペグ配置右 （R の ISL）を設定し、左（L ISL）間のストライドの長さ。 4つの条件は、15センチメートル（SL15）、SLを20％削減を表す対称運動タスクのストライド長（SL）の対称的な制御運動タスクおよび好ましいスピード（SL12）、左肢優先（L9R6）、右が含まれます手足優先（L6R9）運動タスク。 <a href="https://www-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/52921/52921fig1large.jpg" target="_blank">この図の拡大版をご覧になるにはこちらをクリックしてください。</a> <img alt="図2" src="/files/ftp_upload/52921/52921fig2.jpg" /> 非対称性と対角指標の図2.計算。 （A）左肢好ま歩行（L6R9）用姿勢や遊脚期の持続時間（y軸）とサイクル時間（x軸）との関係を回帰分析し、データ点密度のヒートマップにより表されます。位相特性は、傾き切片方程式を用いて、立脚相線形回帰で表されていました。挿入図は、左前肢（LF）、右前肢（RF）、左後肢（LH）および右後肢（RH）ヒートマップに対応する。（B）非対称度算出式に示すように、（1）及び（2）ここで、 R、L、A および p -<html>立脚相線形回帰の傾き、右のために（A）に示すように、左、前方および後方手足、それぞれLAI V、RAIの V、AAI hと PAIの 時間 - 。、垂直左側の縦右、前後方向の水平および後肢水平非対称性指数、それぞれ、 図1で説明したすべての4つの条件について計算（C）対角指数（DIS）式に示すように図1. LF、RF、LH 及び Rhに記載のすべての4つの条件（3）を計算しました。 - 左前肢、右前肢、後肢、右後肢立脚期線形回帰スロープを残した。 <a href="https://www-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/52921/52921fig2large.jpg" target="_blank">この図の拡大版をご覧になるにはこちらをクリックしてください。</a> &quot;&gt; <img alt="図3" src="/files/ftp_upload/52921/52921fig3.jpg" /> すべての8科目から位相特性を用いて、非対称性と対角のために3複合データ図。 （A）スタンスの分布を表すか、左肢好ま歩行（L9R6）のサイクル期間に対するスイングヒートマップ。立脚期の線形回帰の位相特性は、図1（a）のように計算し、傾き切片式差込み図で表されている。 図1（b）に示すように計算された（B）非対称度。ΔlAIV、ΔrAIの V、ΔaAIhおよび ΔpAIH - 、左垂直右、垂直水平前方および後方水平非対称の屈折率差、それぞれ、式（3）に記載のように、対応するasymmを減算することにより、すべての4つの条件について計算左好ま歩行（L9R6）条件から右に好ま歩行（L6R9）のメトリ指数。アスタリスク- 。ブートストラップ法によって計算される統計的有意性は、 <a href="https://www-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/52921/52921fig3large.jpg" target="_blank">この図の拡大版をご覧になるにはこちらをクリックしてください。</a> <img alt="図4" src="/files/ftp_upload/52921/52921fig4.jpg" /> 非対称対策の図4.分析。 （A）。条件L9R6とL6R9間の非対称指数（AI）の絶対差は、複数のテストのためのボンフェローニ補正で調整事後t検定分析を、一方向ANOVAを用いて試験しました。 L9R6とL6R9間の前肢の非対称性（ΔAAIの 時間 ）の変化は有意であった。（B）の分析条件の対角指数（DI）の分布のS15、S12、L9R6及びL6R9は事後T-で一方向ANOVAを用いてのテスト非対称なタスク（黒）との差。 <a href="https://www-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/52921/52921fig4large.jpg" target="_blank">この図の拡大版をご覧になるにはこちらをクリックしてください。</a>

Watch this Scientific Journal Video about Asymmetric Walkway: A Novel Behavioral Assay for Studying Asymmetric Locomotion at JoVE.com

Asymmetric Walkway: A Novel Behavioral Assay for Studying Asymmetric Locomotion

Here, we present a protocol to quantify precise stepping in rodents. Cortical and the spinal central pattern generator signals are required for precise foot-placement during obstructed locomotion. We report here the novel constrained walking task that directly examines precise stepping behavior.

非対称遊歩道：非対称歩行研究のための新しい行動アッセイ

Behavioral assays are commonly used for the assessment of sensorimotor impairment in the central nervous system (CNS). The most sophisticated methods for ...

asymmetric-walkway-novel-behavioral-assay-for-studying-asymmetric

Nanyang Technological University

Research

JoVE Journal

Behavior

8.7K ビュー.  West Virginia University School of Medicine. Here, we present a protocol to quantify precise stepping in rodents. Cortical and the spinal central pattern generator signals are required for precise foot-placement during obstructed locomotion. We report here the novel constrained walking task that directly examines precise stepping behavior.

ビデオ: Asymmetric Walkway: A Novel Behavioral Assay for Studying Asymmetric Locomotion

Behavioral Phenotyping of Murine Disease Models with the Integrated Behavioral Station (INBEST)

Due to rapid advances in genetic engineering, small rodents have become the preferred subjects in many disciplines of biomedical research. In studies of chronic CNS disorders, there is an increasing demand for murine models with high validity at the behavioral level. However, multiple pathogenic mechanisms and complex functional deficits often impose challenges to reliably measure and interpret behavior of chronically sick mice. Therefore, the assessment of peripheral pathology and a behavioral profile at several time points using a battery of tests are required. Video-tracking, behavioral spectroscopy, and remote acquisition of physiological measures are emerging technologies that allow for comprehensive, accurate, and unbiased behavioral analysis in a home-base-like setting. This report describes a refined phenotyping protocol, which includes a custom-made monitoring apparatus (Integrated Behavioral Station, INBEST) that focuses on prolonged measurements of basic functional outputs, such as spontaneous activity, food/water intake and motivated behavior in a relatively stress-free environment. Technical and conceptual improvements in INBEST design may further promote reproducibility and standardization of behavioral studies.

Behavioral Phenotyping of Murine Disease Models with the Integrated Behavioral Station INBEST

Prolonged and comprehensive monitoring of mice in a home-cage environment provides a deeper understanding of aberrant behavior in murine models of brain diseases. This paper describes the Integrated Behavioral Station (INBEST) as the key component of contemporary behavioral analysis.

統合された行動ステーション（INBEST）を有するマウス疾患モデルの行動表現型

Due to rapid advances in genetic engineering, small rodents have become the preferred subjects in many disciplines of biomedical research. In studies of ...

行動学

A Procedure to Observe Context-induced Renewal of Pavlovian-conditioned Alcohol-seeking Behavior in Rats

Environmental contexts in which drugs of abuse are consumed can trigger craving, a subjective Pavlovian-conditioned response that can facilitate drug-seeking behavior and prompt relapse in abstinent drug users. We have developed a procedure to study the behavioral and neural processes that mediate the impact of context on alcohol-seeking behavior in rats. Following acclimation to the taste and pharmacological effects of 15% ethanol in the home cage, male Long-Evans rats receive Pavlovian discrimination training (PDT) in conditioning chambers. In each daily (Mon-Fri) PDT session, 16 trials each of two different 10 sec auditory conditioned stimuli occur. During one stimulus, the CS+, 0.2 ml of 15% ethanol is delivered into a fluid port for oral consumption. The second stimulus, the CS-, is not paired with ethanol. Across sessions, entries into the fluid port during the CS+ increase, whereas entries during the CS- stabilize at a lower level, indicating that a predictive association between the CS+ and ethanol is acquired. During PDT each chamber is equipped with a specific configuration of visual, olfactory and tactile contextual stimuli. Following PDT, extinction training is conducted in the same chamber that is now equipped with a different configuration of contextual stimuli. The CS+ and CS- are presented as before, but ethanol is withheld, which causes a gradual decline in port entries during the CS+. At test, rats are placed back into the PDT context and presented with the CS+ and CS- as before, but without ethanol. This manipulation triggers a robust and selective increase in the number of port entries made during the alcohol predictive CS+, with no change in responding during the CS-. This effect, referred to as context-induced renewal, illustrates the powerful capacity of contexts associated with alcohol consumption to stimulate alcohol-seeking behavior in response to Pavlovian alcohol cues.

A procedure to study the capacity of an alcohol associated environmental context to trigger the renewal of alcohol-seeking behavior in rats is described.

ラットにおけるパブロフ付きアルコール探索行動の状況誘発性のリニューアルを観察する手順

Environmental contexts in which drugs of abuse are consumed can trigger craving, a subjective Pavlovian-conditioned response that can facilitate ...

Performing Behavioral Tasks in Subjects with Intracranial Electrodes

Patients having stereo-electroencephalography (SEEG) electrode, subdural grid or depth electrode implants have a multitude of electrodes implanted in different areas of their brain for the localization of their seizure focus and eloquent areas. After implantation, the patient must remain in the hospital until the pathological area of brain is found and possibly resected. During this time, these patients offer a unique opportunity to the research community because any number of behavioral paradigms can be performed to uncover the neural correlates that guide behavior. Here we present a method for recording brain activity from intracranial implants as subjects perform a behavioral task designed to assess decision-making and reward encoding. All electrophysiological data from the intracranial electrodes are recorded during the behavioral task, allowing for the examination of the many brain areas involved in a single function at time scales relevant to behavior. Moreover, and unlike animal studies, human patients can learn a wide variety of behavioral tasks quickly, allowing for the ability to perform more than one task in the same subject or for performing controls. Despite the many advantages of this technique for understanding human brain function, there are also methodological limitations that we discuss, including environmental factors, analgesic effects, time constraints and recordings from diseased tissue. This method may be easily implemented by any institution that performs intracranial assessments; providing the opportunity to directly examine human brain function during behavior.

Patients implanted with intracranial electrodes provide a unique opportunity to record neurological data from multiple areas of the brain while the patient performs behavioral tasks. Here, we present a method of recording from implanted patients that can be reproducible at other institutions with access to this patient population.

頭蓋内電極を有する被験者に行動タスクの実行

Patients having stereo-electroencephalography (SEEG) electrode, subdural grid or depth electrode implants have a multitude of electrodes implanted in ...

Combined Shuttle-Box Training with Electrophysiological Cortex Recording and Stimulation as a Tool to Study Perception and Learning

Shuttle-box avoidance learning is a well-established method in behavioral neuroscience and experimental setups were traditionally custom-made; the necessary equipment is now available by several commercial companies. This protocol provides a detailed description of a two-way shuttle-box avoidance learning paradigm in rodents (here Mongolian gerbils; Meriones unguiculatus) in combination with site-specific electrical intracortical microstimulation (ICMS) and simultaneous chronical electrophysiological in vivo recordings. The detailed protocol is applicable to study multiple aspects of learning behavior and perception in different rodent species.
Site-specific ICMS of auditory cortical circuits as conditioned stimuli here is used as a tool to test the perceptual relevance of specific afferent, efferent and intracortical connections. Distinct activation patterns can be evoked by using different stimulation electrode arrays for local, layer-dependent ICMS or distant ICMS sites. Utilizing behavioral signal detection analysis it can be determined which stimulation strategy is most effective for eliciting a behaviorally detectable and salient signal. Further, parallel multichannel-recordings using different electrode designs (surface electrodes, depth electrodes, etc.) allow for investigating neuronal observables over the time course of such learning processes. It will be discussed how changes of the behavioral design can increase the cognitive complexity (e.g. detection, discrimination, reversal learning).

Shuttle-box avoidance learning is well-established in behavioral neuroscience. This protocol describes how shuttle-box learning in rodents can be combined with site-specific electrical intracortical microstimulation (ICMS) and simultaneous chronical in vivo recordings as a tool to study multiple aspects of learning and perception.

知覚と学習を研究するためのツールと​​しての電気生理学的皮質の記録と刺激と組み合わせたシャトル・ボックストレーニング

Shuttle-box avoidance learning is a well-established method in behavioral neuroscience and experimental setups were traditionally custom-made; the ...

Novel Object Exploration as a Potential Assay for Higher Order Repetitive Behaviors in Mice

Restricted, repetitive behaviors (RRBs) are a core feature of autism spectrum disorder (ASD) and disrupt the lives of affected individuals. RRBs are commonly split into lower-order and higher-order components, with lower order RRBs consisting of motor stereotypies and higher order RRBs consisting of perseverative and sequencing behaviors. Higher order RRBs are challenging to model in mice. Current assays for RRBs in mice focus primarily on the lower order components, making basic biomedical research into potential treatments or interventions for higher-order RRBs difficult. Here we describe a new assay, novel object exploration. This assay uses a basic open-field arena with four novel objects placed around the perimeter. The test mouse is allowed to freely explore the arena and the order in which the mouse investigates the novel objects is recorded. From these data, patterned sequences of exploration can be identified, as can the most preferred object for each mouse. The representative data shared here and past results using the novel object exploration assay illustrate that inbred mouse strains do demonstrate different behavior in this assay and that strains with elevated lower order RRBs also show elevated patterned behavior. As such, the novel object exploration assay appears to possess good face validity for higher order RRBs in humans and may be a valuable assay for future studies investigating novel therapeutics for ASD.

Higher order restricted, repetitive behaviors (RRBs) disrupt the lives of affected individuals. These behaviors are challenging to model in rodents, making basic biomedical research into potential treatments or interventions for RRBs difficult. Here we describe novel object exploration as a potential assay for higher order RRBs in mice.

マウスにおける高次反復行動のための電位アッセイとして新規物体探査

Restricted, repetitive behaviors (RRBs) are a core feature of autism spectrum disorder (ASD) and disrupt the lives of affected individuals.  RRBs are ...

The Rodent Psychomotor Vigilance Test (rPVT): A Method for Assessing Neurobehavioral Performance in Rats and Mice

The human Psychomotor Vigilance Test (PVT) is a widely used procedure for measuring changes in fatigue and sustained attention. The present article describes a rodent version of the PVT&#8212;termed the &#34;rPVT&#34;&#8212;that measures similar aspects of attention (i.e., performance accuracy, motor speed, premature responding, and lapses in attention). Data are presented that demonstrate both the short- and long-term usefulness of the rPVT when employed with laboratory rats. Rats easily learn the rPVT, and learning to perform the basic procedure takes less than two weeks of training. Once acquired, rat performances in the rPVT show a high degree of similarity to these same performance measures in the human PVT, including similarities in, lapses in attention, reaction times, vigilance decrements across session time (i.e., the human &#34;time-on-task&#34; effects), and the response-stimulus interval (RSI) effect described for humans. Thus the rPVT can be an extremely valuable tool for assessing the effects of a wide range of variables on sustained attention quite similar to human PVT performances, and thus can be useful for developing novel treatments for neurobehavioral dysfunctions.

The Rodent Psychomotor Vigilance Test rPVT: A Method for Assessing Neurobehavioral Performance in Rats and Mice

A rat version of the human Psychomotor Vigilance Test (PVT) is described that measures aspects of attention similar to those measured with the human PVT, including aspects of human vigilance such as performance accuracy, motor speed, premature responding, and lapses in attention.

げっ歯類精神運動警戒試験（rPVT）：ラットとマウスの神経行動のパフォーマンスを評価するための方法

The human Psychomotor Vigilance Test (PVT) is a widely used procedure for measuring changes in fatigue and sustained attention. The present article ...

Battery of Behavioral Tests Assessing General Locomotion, Muscular Strength, and Coordination in Mice

Behavioral testing is used in pre-clinical trials to assess the phenotypic effects and outcomes that a particular disease or treatment has on the animal&#39;s wellbeing and health. There are numerous behavioral tests that may be applied. We selected a test for general locomotion, the open field test (OFT); a test for muscular strength, the mesh test (MT); and a test for coordination, the rotarod test (RR). Testing can be accomplished on a weekly or monthly basis. As a test for general locomotion, the OFT works by objectively monitoring movement parameters while the mouse is in an open field apparatus. The field is generally a 2&#39; x 2&#39; box, and the movements are recorded through laser sensing or through video capture. The mouse is placed in the center of the open field and allowed to move freely for the test. The MT uses the latency for a mouse to fall off an inverted screen as a measure of muscular strength. A mouse is placed on a screen, which is inverted over a clear box, and is timed for their latency to fall. Three trials are performed, with the best of the three trials scored for that day. A score of 60 s is the maximum time a mouse is left inverted. Mice are given a 5-min rest period between mesh test trials. Lastly, an accelerated protocol on the RR assesses motor coordination and endurance. During a trial, a mouse walks on a rotating rod as it increases in speed from 4 rpm to 40 rpm over 5 min. The trial ends when the mouse touches the magnetized pressure sensor upon falling. Each mouse undergoes three trials, and the best trial is scored for that day. This combined behavioral data allows for the global assessment of mobility, coordination, strength, and movement of the test animals. At least two out of the three behavioral testing measures must show improvement for an animal to qualify as having overall improved motor function.

Behavioral testing is used in pre-clinical trials to assess the phenotypic effects of a disease or treatment on an animal's wellbeing. In order to globally assess motor functioning, we selected tests for general locomotion, muscular strength, and coordination: the open field test, the mesh test, and the rotarod test, respectively.

行動のバッテリー テスト評価の一般的な運動、筋肉の強さと連携マウス

Behavioral testing is used in pre-clinical trials to assess the phenotypic effects and outcomes that a particular disease or treatment has on the ...

A Wireless, Bidirectional Interface for In Vivo Recording and Stimulation of Neural Activity in Freely Behaving Rats

In vivo electrophysiology is a powerful technique to investigate the relationship between brain activity and behavior at a millisecond and micrometer scale. However, current methods mostly rely on tethered cable recordings or only use unidirectional systems, allowing either recording or stimulation of neural activity, but not at the same time or same target. Here, a new wireless, bidirectional device for simultaneous multichannel recording and stimulation of neural activity in freely behaving rats is described. The system operates through a single portable head stage that both transmits recorded activity and can be targeted in real-time for brain stimulation using a telemetry-based multichannel software. The head stage is equipped with a preamplifier and a rechargeable battery, allowing stable long-term recordings or stimulation for up to 1 h. Importantly, the head stage is compact, weighs 12 g (including battery) and thus has minimal impact on the animal&#180;s behavioral repertoire, making the method applicable to a broad set of behavioral tasks. Moreover, the method has the major advantage that the effect of brain stimulation on neural activity and behavior can be measured simultaneously, providing a tool to assess the causal relationships between specific brain activation patterns and behavior. This feature makes the method particularly valuable for the field of deep brain stimulation, allowing precise assessment, monitoring, and adjustment of stimulation parameters during long-term behavioral experiments. The applicability of the system has been validated using the inferior colliculus as a model structure.

A Wireless, Bidirectional Interface for In Vivo Recording and Stimulation of Neural Activity in Freely Behaving Rats

A wireless, bidirectional system for multi-channel neural recordings and stimulation in freely behaving rats is introduced. The system is light and compact, thus having minimal impact on the animal&#180;s behavioral repertoire. Moreover, this bidirectional system provides a sophisticated tool to assess causal relationships between brain activation patterns and behavior.

ワイヤレス、 In Vivo記録そして自由行動下ラットにおける神経活動の刺激のための双方向インタ フェース

In vivo electrophysiology is a powerful technique to investigate the relationship between brain activity and behavior at a millisecond and micrometer ...

A Behavioral Assay for Investigating the Role of Spatial Memory During Instinctive Defense in Mice

Evolution has selected a repertoire of defensive behaviors that are essential for survival across all animal species. These behaviors are often stereotyped actions elicited in response to innately aversive sensory stimuli, but their success requires enough flexibility for adapting to different spatial environments, which can change rapidly. Here, we describe a behavioral assay to evaluate the influence of learned spatial knowledge on defensive behaviors in mice. We have adapted the widely used Barnes maze spatial memory assay to investigate how mice navigate to a shelter during escape responses to innately aversive sensory stimuli in a novel environment, and how they adapt to acute changes in the environment. This new assay is an ethological paradigm that does not require training and exploits the natural exploration patterns and navigation strategies in mice. We propose that the set of protocols described here are a powerful means of studying goal-directed behaviors and stimulus-triggered navigation, which should be of interest to both the fields of instinctive behaviors and spatial memory.

This protocol describes a behavioral assay, based on the Barnes maze test, to study how instinctive defensive actions are modified by the knowledge of spatial environment.

マウスで本能的な防衛中に空間記憶の役割を調査するため行動分析

Evolution has selected a repertoire of defensive behaviors that are essential for survival across all animal species. These behaviors are often ...

Automated, Long-term Behavioral Assay for Cognitive Functions in Multiple Genetic Models of Alzheimer's Disease, Using IntelliCage

Multiple factors&#8212;such as aging and genes&#8212;are frequently associated with cognitive decline. Genetically modified mouse models of cognitive decline, such as Alzheimer&#39;s disease (AD), have become a promising tool to elucidate the underlying mechanisms and promote the therapeutic advances. An important step is the validation and characterization of expected behavioral abnormality in the models, in the case of AD, cognitive decline. The long-term behavioral investigations of laboratory animals to study the effect of aging demand substantial efforts from researchers. The IntelliCage system is a high-throughput and cost-effective test battery for mice that eliminates the need for daily human handling. Here, we describe how the system is utilized in the long-term phenotyping of a genetic Alzheimer&#39;s disease model, specifically focusing on the cognitive functions. The experiment employs repeated battery of tests that assess spatial learning and executive functions. This cost-effective age-dependent phenotyping allows us to identify the transient and/or permanent effects of genes on various cognitive aspects.

This paper describes a protocol for cognitive assessments for genetic models of the Alzheimer&#39;s disease using the IntelliCage system, which is a high throughput automated behavioral monitoring system with operant conditioning.

アルツハイマー病、IntelliCage を使用しての複数の遺伝モデルにおける認知機能の長期的な行動のアッセイの自動化

Multiple factors&#8212;such as aging and genes&#8212;are frequently associated with cognitive decline. Genetically modified mouse models of cognitive ...