この作業によって、国家機関の健康/国立研究所の神経疾患をサポートされていたし、ストロークを与える 1R01NS096225 01A1、アメリカの心臓協会許可 AHA 13SDG1395001413、AHA 17GRNT33660336、AHA-17POST33660174、ルイジアナ州立大学助成援助研究評議会、マルコム ・ ファイスト心血管研究フェローシップ、およびエベリン F. マクナイト脳研究所で。

すべて実験プロシージャが健康の国民の協会のガイドラインに従って行われ、機関動物ケアおよび使用委員会 (ルイジアナ州立大学健康科学センター-シュリーブ ポート) 男性のスプレイグ ドーリー ラット (の使用が承認300-350 g、9-10 週齢)。ラットが ACA 手術前に一晩絶食。
1. における装置の設計と設定
注: の執事およびローリンズの 2006年モデル31基におけるデザイン。
<ol><li>SketchUp32を利用した迷路、3 D 構造を設計します。におけるの 3 D 構造を作成するには、外長さ 200 mm、幅 165 mm、高さ 148 mm 3 D プリンターの印刷寸法に合わせて開始腕を構築します。肉厚 5.5 mm と 8 mm の床厚を使用します。</li>
	<li>3 D プリンターを使用して迷路を印刷 (材料の表参照)32。研究室では、3 D プリンターが一覧にない場合は、木、媒体密度の有機質繊維板、または改築の店から購入することができますプラスチック (ポリ塩化ビニールすなわち) など他の材料を使用します。<ol>
<li>印刷領域の高さ制限のため 2 つの独立した 3 D プリントの迷路の壁を構築し、迷路アセンブリに結合 (つまり、2 番目の壁の高さが 140 mm、280 mm の総壁の高さによって高さを増加する迷路セクションに追加された)。それぞれ独立した 3 D 印刷拠点には、"T"の形のロック機構は、1 つのセクションが、次に接続が含まれています。</li>
		<li>開始の目標腕くんでの交差点、その目標腕開始アームの幅に参加する 165 mm の広いセクションを作成します。開始腕として同じような設計法を用いた目標武器を構築します。ただし、執事およびローリンズのデザインごと 100 mm アームの幅を減らします。</li>
		<li>におけるの回路図/寸法の詳細につきましては、図 1を参照してください。</li>
		<li>開始腕と目標腕の接合部で設計に中央のパーティションを含んでいます。目標の腕に分割する開始の腕に、におけるのと 200 mm 後ろの壁からこのパーティションを拡張します。このパーティションはまた迷路 (図 1) の高さを拡張します。</li>
	</ol>
</li>
</ol>2. 仮死心不全 (ACA)
<ol><li>手術の開始前にオートクレーブの手術器具の (121 ° C で 15 分)。手術のサイトで動物の毛 70% のエタノール 15 分ひげをそるが手術台を消毒します。外科手術のための表面の皮を betadine ソリューションを適用します。</li>
	<li>Anesthetization
	<ol>
<li>4% イソフルランと無段階の混合物と O2と N2O (300 mL/分 O2と 700 mL/分 N2O) マスクを介してラットを麻酔します。</li>
		<li>与えるラット換気用気管内挿管 (挿管後ラットが人工呼吸器に接続されて)。</li>
		<li>O2と N2O を使用ピンチ応答法麻酔の深さを決定するための無段階の混合物 2% に 4% からイソフルランを下げることによって麻酔を維持します。</li>
		<li>麻酔中の乾燥を防ぐための目に軟膏を塗る。Regulae 加熱温度基準として肛門プローブ パッド齧歯動物によって体の温度。</li>
	</ol></li>
	<li>気管内挿管
	<ol>
<li>誘導室にネズミを配置します。4% のイソフルランと無段階の混合物の O2と N2o. ラットを麻酔します。</li>
		<li>誘導室からラットを削除します。場所は、ネズミの顔に向かって麻酔マスクで仰臥位で動物を麻酔しました。</li>
		<li>そっと左の親指と人差し指の動物の左右どちらかに向けて舌を移動します。</li>
		<li>17 ゲージ鈍先端ピペッティング針 (93 mm の長さの針の先端に 10 度の角度で) 上 14 ゲージ フレキシブル静脈カテーテル (49 mm 長い) を滑空します。17 ゲージ鈍チップ分注針を気管に挿入します。</li>
		<li>気管から 17 g の分注針をそっと引き出します。14 ゲージ カテーテルのハブを人工呼吸器に接続します。0.67 mL/100 g と 60 呼吸/分の呼吸を人工呼吸器ストローク ボリュームを調整します。</li>
		<li>加熱温度基準として肛門プローブ パッド齧歯動物によって全体の手順中に 37 ° C で頭と体の温度を維持します。</li>
	</ol></li>
	<li>大腿動脈と静脈のカテーテル法
	<ol>
<li>(両側) 鼠径部付近の毛を剃るし、皮膚の外科手術のための表面に betadine を適用します。</li>
		<li>ラットを仰臥位に配置されます。手術用ハサミで、鼠径部の切開 (10 mm) を行います。</li>
		<li>鼠径靭帯の公開まで結合組織を鈍先端の鉗子に区切ります。鼠径靭帯を把握するため、止血を使用します。大腿動脈および静脈は、鼠径靱帯の下です。</li>
		<li>大腿動脈および静脈が公開されるまでは、結合組織を分離するのに鈍い先端の鉗子を使用します。</li>
		<li>ファインチップ鉗子を介して大腿動脈に沿って実行される大腿神経を優しきます。慎重に大腿動脈を分離、ファインチップ鉗子を介してユニットとして静脈。</li>
		<li>ファインチップ鉗子を使用して、静脈から大腿動脈を区切ります。</li>
		<li>5-0 絹糸 (1 脚と体の方へ他の方) 静脈の下の 2 個を配置します。</li>
		<li>体に近い側に緩やかな結び目を作る。保持し、体の反対側に向かって可能な限り引っ張って、止血を使用します。</li>
		<li>脚に近い側の緩い結び目を作る。押しながら静脈血でいっぱいにできるように止血を介して足の方に引っ張ってください。</li>
		<li>(45 ° の角度) でミクロ解剖はさみによって静脈 (約 0.1 mm) の小切開を作る。滅菌ガーゼで血を吸収します。</li>
		<li>PE 50 カテーテル (20 U/mL と生理食塩水で満ちている) 鈍い先端針注射器に接続します。20 U/mL と生理食塩水で PE 50 カテーテルを入力します。解剖はさみで PE 50 カテーテルをポイントを作成または鋭い端を 45 ° の角度でカットします。PE 50 カテーテルの端を保持するのに鈍い先端の鉗子を使用します。そっと PE 50 カテーテルを大腿静脈に挿入します。</li>
		<li>カテーテルを完全に挿入すると後は、ゆっくりと 0.1 mL のヘパリン/食塩水漏れがないことを確認するを管理します。PE 50 カテーテルを安定させるためにしっかり縫合結び目 (シングル ノット) を結ぶ。様々 な薬の連続投与 (IV) 用 PE 50 カテーテルをしてください。</li>
		<li>1 mL 注射器で使用、23 ゲージのプロシージャ全体でラットを動けなく大腿静脈経由でルアー スタブ アダプター ベクロニウム臭化物 (0.67 mg/kg、10 分毎に投与) を管理します。</li>
		<li>5-0 絹糸 (1 脚と体の方へ他の方)、動脈の下の 2 個を配置します。</li>
		<li>脚に近い側の緩い結び目を作る。脚に向かって可能な限り縫合をプルして、止血を使用します。</li>
		<li>体に近い側に緩やかな結び目を作る。動脈の血液で充満を許可する止血を介して体の方に引っ張って押しします。</li>
		<li>(45 ° の角度) でミクロ解剖はさみで動脈 (約 0.1 mm) の小切開を作る。</li>
		<li>PE 50 カテーテル (20 U/mL と生理食塩水で満ちている) 鈍い先端針注射器に接続します。20 U/mL と生理食塩水で PE 50 カテーテルを入力します。解剖はさみで PE 50 カテーテルをポイントまたは鋭い端を作成する 45 度の角度でカットします。PE 50 カテーテルの端を保持するのに鈍い先端の鉗子を使用します。PE 50 カテーテルの端を保持するのに鈍い先端の鉗子を使用します。優しく PE 50 カテーテルを大腿動脈に挿入します。</li>
		<li>後カテーテルを完全に挿入すると、ゆっくりと引くそのカテーテルが機能ように注射器。PE 50 カテーテルを安定させるためにしっかり縫合結び目 (シングル ノット) を結ぶ。PE 50 カテーテル動脈圧と血液ガス連続記録を保持します。</li>
	</ol></li>
	<li>仮死の心不全 (ACA) 手順
	<ol>
<li>ストローク量、O2 N2O のレベルを調節することにより、必要に応じて (つまり pO2pCO2血圧、および pH 値) 生理学的パラメーターを調整します。これらのパラメーターの正常な生理学的な範囲を使用: pO2: 100 mmHg、pCO2: 35 40 mmHg、血圧: 100 mmHg と pH: 7.4。</li>
		<li>ACA を実行する前に約 100 mmHg または大腿静脈経由でベクロニウム臭化物 (0.67 mg/kg、静注) を管理し、2 分待つ 23 ゲージ ルアー スタブ アダプターを接続して 1 mL 注射器、血圧は、ことを確認使用します。</li>
		<li>人工呼吸器から気管内チューブ (14 ゲージ カテーテル ハブ) を切断することによって無呼吸 (6 分) を誘発します。さらに完全な無呼吸を確保するための 1 mL シリンジで気管内チューブをブロックします。 注: 6 分仮死時間人工呼吸器切断と蘇生の初め間の期間として定義されます。完全な心停止は、平均動脈圧 10 mmHg より低いとして定義されます。</li>
		<li>無呼吸の最後の分の間に 80 の呼吸/分、人工呼吸器の呼吸を調整し、O2 0% N2O を 2 L/分に増加この操作は、任意の残りのイソフルランと N2O は、人工呼吸器の残りが爆破されます。</li>
		<li>次の無呼吸の分は、気管内チューブから 1 mL の注射器を削除します。人工呼吸器を気管内チューブを接続し直します。</li>
		<li>ルアー スタブ アダプター大腿静脈からエピネフリン (0.005 mg/kg、静注) を管理でき x 光円運動で動物の胸に指と親指、中指、によって手動の胸部圧迫を管理するゲージを使用、23 の 1 mL 注射器が接続されていると自然循環 (平均動脈圧 ≥ 50 mmHg)33,34,35のリターンまでの z 軸 (200/分) です。</li>
		<li>使用別 1 mL 注射器、23 ゲージ自然循環へのリターンの直後に大腿静脈経由で重炭酸ナトリウム 1 meq/kg、静注) を管理するルアー スタブ アダプター (50 mmHg 以上)33,34,35呼吸性アシドーシスを軽減するために。</li>
		<li>測定血液ガス再度 10 分蘇生 (pH ACA 約 7.35 に 7.40 後) の酸塩基状態を確認後</li>
		<li>大腿動脈および静脈を揃えるため、止血を使用します。ゆっくりと優しく鈍い先端の鉗子を用いた動脈と静脈のカテーテルを削除します。大腿動脈/静脈の出血を防ぐために 5-0 絹糸を結紮します。3-0 絹糸を使用して手術部位を覆う皮膚を閉じます。中断縫合テクニックを使用すると、傷の再開の可能性を最小限に抑えます。</li>
		<li>ラット呼吸自体 (蘇生後 60 分通常 30 分) まで待ち、ベンチレーターからラットを切断、気管内チューブを慎重に取り外します。</li>
		<li>場所赤ちゃんインキュベーター (27 ° C、湿度 50%) でラットの一夜します。(水に浸漬製) 軟化食品を置き、赤ん坊の定温器に一晩水します。</li>
		<li>個々 のケージにラットを転送し、通常の食事と水の動物施設にラットを返します。におけるテストは、ACA の後 3 日間を開始します。</li>
	</ol></li>
</ol>3. における
<ol><li>
動物の準備
	<ol>
<li>(偽または ACA) は、手術の前日は、5 分ごとのラットを処理します。決してとき彼らのおり (480 mm × 250 mm × 200 mm、プラスチックの透明ケージ) からラットを昇格処理 (図 2)。</li>
		<li>ラットを処理した後、そっと拾うラット他の手のサポートで片手で尻尾でその ' 足。5 倍 (100 mm) ケージに手からジャンプさせてください。彼らは食糧および/または戦いの支配していないので、個々 のケージに各ラットを分けます。</li>
		<li>シャムや ACA 手術 (図 2) 後、3 日間は、最初の実行の開始前に静かで暗い部屋にケージでラットを転送します。だけ低消費電力デスク ランプをオンに、最低被写体照度を維持するために実験室の隅に置きます。10 分闇に適応するラットを許可します。</li>
		<li>午後にはラットのパフォーマンスに日内変動の効果を避けるためにすべての実験を実行します。オペレーターは受信したラット偽または ACA 手術がお勧めしません。</li>
	</ol>
</li>
	<li>
自発交替
	<ol>
<li>迷路の全体の床をカバーする (〜 10 mm 厚) を寝具の薄い層を広げます。それぞれの出発点です開始腕 ("T"の下) でラットの実行、および許可し、場所はラット右または左の目標腕を探索する 3 分です。</li>
		<li>ラット (ネズミのすべての 4 つの足は目標腕にした) 特定の目標腕にコミットされると、"T"接合開始腕と反対の目標腕ラットが反対の目標腕を入力できないようにする (図 1) はブロックします。30 秒間、迷路の中でネズミを残すし、ラットをピックアップし、最小限の時間 (~ 30 秒) についてそのケージに戻ってそれを配置します。"T"ジャンクションを削除、におけるから (125 mm X 230 mm X 65 mm、3 D プリンターによって作られた) をブロックします。</li>
		<li>開始腕にラットを置き、3.2.2 を繰り返します。交替として定義されて: ラットと比較して36を実行前の反対側の腕に入ったとき。1 日あたりの実行の通りを 4 ラットがあります。 1stを実行 2ndを実行します。 10 分休憩 3rdを実行します。 4回実行します。</li>
		<li>10 分休憩の間に、香りのバイアスを除去するために動物の寝具を変更します。における実験的毎日の終わりに蒸留水が続く 75% エタノールできれいに。</li>
		<li>3.2.1 の手順を繰り返します。-3.2.4。図 2のように 2 つ以上日 (合計 12 の実行)。</li>
	</ol>
</li>
	<li>
交代率と側優先率の計算
	<ol>
<li>交替 % と % 側の好みが、計算、 L: ラットが左の腕を選択します。 R: ラットを選択右の腕 正解: (各セットは、2 つの実行を含んでいる) 与えられたセットの 1stと 2ndを実行は違います 不適切な選択: ラットが以前の実行と同様同じ腕を選択 <img alt="Equation 1" src="/files/ftp_upload/56694/56694eq1.jpg"> <img alt="Equation 2" src="/files/ftp_upload/56694/56694eq2.jpg"> 例: 1 日目: L L/L 2 日目: L L/L R 3 日目: R L/L 交代: 2 (正しい選択肢) 6 (合計セット実行) * 100 = 33.33% 側の好み: 10 (L、優先側) 12 (合計実行実行) * 100 = 83.33%</li>
	</ol>
</li>
	<li>
手術後のケア:
	<ol>
<li>与えるは、手術後 2 日間 12 時間間隔ブプレノルフィン (0.01 mg/kg IP) をラットします。心停止後 1 h までのラットを観察します。</li>
		<li>それは胸骨の横臥を維持するために十分な意識を取り戻したまで人工呼吸器や加熱パッドにラットを取り付けます。術後後の動物の体温を維持するために (27 ° c、湿度 50% 設定) 赤ちゃんのインキュベーターでラットを配置します。</li>
		<li>手術後最初の 24 時間の動物に軟化チョウ (水浸漬製) を提供します。ラットの飲料水がなかったら場合、管理静菌性生理食塩水 (100 mL/kg/日、i. p.) まで動物が回復し、自由に水を飲むのです。</li>
		<li>すべての傷に痛み (バシトラシンとリドカイン軟膏) をラットの局所用抗生物質を与えます。完全に回復した後、ラットを動物施設に戻って移動します。</li>
	</ol>
</li>
	<li>
安楽死の方法
	<ol>
<li>5% イソフルランと 100% N2O は、実験の終わりに動物の安楽死を使用します。</li>
	</ol>
</li>
</ol>

<ol>
	<li>Writing Group, M., 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=Heart+Disease+and+Stroke+Statistics-2016+Update:+A+Report+From+the+American+Heart+Association.">Heart Disease and Stroke Statistics-2016 Update: A Report From the American Heart Association.</a> Circulation. 133, e38-e360 (2016).</li><li>Beuret, P., 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=Cardiac+arrest:+prognostic+factors+and+outcome+at+one+year.">Cardiac arrest: prognostic factors and outcome at one year.</a> Resuscitation. 25, 171-179 (1993).</li><li>Kim, Y. J., 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=Long-term+neurological+outcomes+in+patients+after+out-of-hospital+cardiac+arrest.">Long-term neurological outcomes in patients after out-of-hospital cardiac arrest.</a> Resuscitation. 101, 1-5 (2016).</li><li>Earnest, M. P., Yarnell, P. R., Merrill, S. L., Knapp, G. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Long-term+survival+and+neurologic+status+after+resuscitation+from+out-of-hospital+cardiac+arrest.">Long-term survival and neurologic status after resuscitation from out-of-hospital cardiac arrest.</a> Neurology. 30, 1298-1302 (1980).</li><li>Cerchiari, E. L., Safar, P., Klein, E., Cantadore, R., Pinsky, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cardiovascular+function+and+neurologic+outcome+after+cardiac+arrest+in+dogs.+The+cardiovascular+post-resuscitation+syndrome.">Cardiovascular function and neurologic outcome after cardiac arrest in dogs. The cardiovascular post-resuscitation syndrome.</a> Resuscitation. 25, 9-33 (1993).</li><li>Petito, C. K., Feldmann, E., Pulsinelli, W. A., Plum, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Delayed+hippocampal+damage+in+humans+following+cardiorespiratory+arrest.">Delayed hippocampal damage in humans following cardiorespiratory arrest.</a> Neurology. 37, 1281-1286 (1987).</li><li>Schmidt-Kastner, R., Freund, T. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Selective+vulnerability+of+the+hippocampus+in+brain+ischemia.">Selective vulnerability of the hippocampus in brain ischemia.</a> Neuroscience. 40, 599-636 (1991).</li><li>Corkin, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=What's+new+with+the+amnesic+patient+H.M.?.">What's new with the amnesic patient H.M.?.</a> Nat Rev Neurosci. 3, 153-160 (2002).</li><li>Scoville, W. B., Milner, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Loss+of+recent+memory+after+bilateral+hippocampal+lesions.+1957.">Loss of recent memory after bilateral hippocampal lesions. 1957.</a> J Neuropsychiatry Clin Neurosci. 12, 103-113 (2000).</li><li>Smith, T. D., Calhoun, M. E., Rapp, P. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Circuit+and+morphological+specificity+of+synaptic+change+in+the+aged+hippocampal+formation.">Circuit and morphological specificity of synaptic change in the aged hippocampal formation.</a> Neurobiol Aging. 20, 357-358 (1999).</li><li>Gage, F. H., Dunnett, S. B., Bjorklund, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Spatial+learning+and+motor+deficits+in+aged+rats.">Spatial learning and motor deficits in aged rats.</a> Neurobiol Aging. 5, 43-48 (1984).</li><li>Tulving, E., Markowitsch, H. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Episodic+and+declarative+memory:+role+of+the+hippocampus.">Episodic and declarative memory: role of the hippocampus.</a> Hippocampus. 8, 198-204 (1998).</li><li>Neigh, G. N., 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=Cardiac+arrest+with+cardiopulmonary+resuscitation+reduces+dendritic+spine+density+in+CA1+pyramidal+cells+and+selectively+alters+acquisition+of+spatial+memory.">Cardiac arrest with cardiopulmonary resuscitation reduces dendritic spine density in CA1 pyramidal cells and selectively alters acquisition of spatial memory.</a> Eur J Neurosci. 20, 1865-1872 (2004).</li><li>Volpe, B. T., Davis, H. P., Towle, A., Dunlap, W. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Loss+of+hippocampal+CA1+pyramidal+neurons+correlates+with+memory+impairment+in+rats+with+ischemic+or+neurotoxin+lesions.">Loss of hippocampal CA1 pyramidal neurons correlates with memory impairment in rats with ischemic or neurotoxin lesions.</a> Behav Neurosci. 106, 457-464 (1992).</li><li>Astur, R. S., Taylor, L. B., Mamelak, A. N., Philpott, L., Sutherland, R. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Humans+with+hippocampus+damage+display+severe+spatial+memory+impairments+in+a+virtual+Morris+water+task.">Humans with hippocampus damage display severe spatial memory impairments in a virtual Morris water task.</a> Behav Brain Res. 132, 77-84 (2002).</li><li>Lee, R. H., 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=Fatty+acid+methyl+esters+as+a+potential+therapy+against+cerebral+ischemia.">Fatty acid methyl esters as a potential therapy against cerebral ischemia.</a> OCL. 23, D108 (2016).</li><li>Lee, R. H., Porto, L. F., 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=Chapter+1.">Chapter 1.</a> Palmitic acid: occurrence, biochemistry and health effects. , 1-15 (2014).</li><li>Tolman, E. C., Gleitman, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Studies+in+spatial+learning;+place+and+response+learning+under+different+degrees+of+motivation.">Studies in spatial learning; place and response learning under different degrees of motivation.</a> J Exp Psychol. 39, 653-659 (1949).</li><li>Koopmans, G., Blokland, A., van Nieuwenhuijzen, P., Prickaerts, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Assessment+of+spatial+learning+abilities+of+mice+in+a+new+circular+maze.">Assessment of spatial learning abilities of mice in a new circular maze.</a> Physiol Behav. 79, 683-693 (2003).</li><li>Barnes, C. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Memory+deficits+associated+with+senescence:+a+neurophysiological+and+behavioral+study+in+the+rat.">Memory deficits associated with senescence: a neurophysiological and behavioral study in the rat.</a> J Comp Psychol. 93, 74-104 (1979).</li><li>Paul, C. M., Magda, G., Abel, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Spatial+memory:+Theoretical+basis+and+comparative+review+on+experimental+methods+in+rodents.">Spatial memory: Theoretical basis and comparative review on experimental methods in rodents.</a> Behav Brain Res. 203, 151-164 (2009).</li><li>Vorhees, C. V., Williams, M. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Assessing+spatial+learning+and+memory+in+rodents.">Assessing spatial learning and memory in rodents.</a> ILAR J. 55, 310-332 (2014).</li><li>Sharma, S., Rakoczy, S., Brown-Borg, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Assessment+of+spatial+memory+in+mice.">Assessment of spatial memory in mice.</a> Life Sci. 87, 521-536 (2010).</li><li>Poon, T. P., 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=Spinal+cord+toxoplasma+lesion+in+AIDS:+MR+findings.">Spinal cord toxoplasma lesion in AIDS: MR findings.</a> J Comput Assist Tomogr. 16, 817-819 (1992).</li><li>Sunyer, B., Patil, S., H&#246;ger, H., Lubec, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Barnes+maze,+a+useful+task+to+assess+spatial+reference+memory+in+the+mice.">Barnes maze, a useful task to assess spatial reference memory in the mice.</a> Nat Protoc. 390, (2007).</li><li>Shoji, H., Hagihara, H., Takao, K., Hattori, S., Miyakawa, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=T-maze+forced+alternation+and+left-right+discrimination+tasks+for+assessing+working+and+reference+memory+in+mice.">T-maze forced alternation and left-right discrimination tasks for assessing working and reference memory in mice.</a> J Vis Exp. , (2012).</li><li>Seeger, T., 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=M2+muscarinic+acetylcholine+receptor+knock-out+mice+show+deficits+in+behavioral+flexibility,+working+memory,+and+hippocampal+plasticity.">M2 muscarinic acetylcholine receptor knock-out mice show deficits in behavioral flexibility, working memory, and hippocampal plasticity.</a> J Neurosci. 24, 10117-10127 (2004).</li><li>Olton, D. S., Feustle, W. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hippocampal+function+required+for+nonspatial+working+memory.">Hippocampal function required for nonspatial working memory.</a> Exp Brain Res. 41, 380-389 (1981).</li><li>Hayashida, K., 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=Hydrogen+inhalation+during+normoxic+resuscitation+improves+neurological+outcome+in+a+rat+model+of+cardiac+arrest+independently+of+targeted+temperature+management.">Hydrogen inhalation during normoxic resuscitation improves neurological outcome in a rat model of cardiac arrest independently of targeted temperature management.</a> Circulation. 130, 2173-2180 (2014).</li><li>Dember, W. N., Richman, C. 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> Spontaneous alternation behavior. , (1989).</li><li>Deacon, R. M., Rawlins, J. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=T-maze+alternation+in+the+rodent.">T-maze alternation in the rodent.</a> Nature protocols. 1, 7-12 (2006).</li><li>Wong, K. V., Hernandez, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+review+of+additive+manufacturing.">A review of additive manufacturing.</a> ISRN Mechanical Engineering. 2012, (2012).</li><li>Lin, H. W., 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=Derangements+of+post-ischemic+cerebral+blood+flow+by+protein+kinase+C+delta.">Derangements of post-ischemic cerebral blood flow by protein kinase C delta.</a> Neuroscience. 171, 566-576 (2010).</li><li>Lin, H. W., 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=Fatty+acid+methyl+esters+and+Solutol+HS+15+confer+neuroprotection+after+focal+and+global+cerebral+ischemia.">Fatty acid methyl esters and Solutol HS 15 confer neuroprotection after focal and global cerebral ischemia.</a> Transl Stroke Res. 5, 109-117 (2014).</li><li>Lee, R. H., 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=Interruption+of+perivascular+sympathetic+nerves+of+cerebral+arteries+offers+neuroprotection+against+ischemia.">Interruption of perivascular sympathetic nerves of cerebral arteries offers neuroprotection against ischemia.</a> Am J Physiol Heart Circ Physiol. 312, H182-H188 (2017).</li><li>Bali, Z. K., 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=Differential+effects+of+alpha7+nicotinic+receptor+agonist+PHA-543613+on+spatial+memory+performance+of+rats+in+two+distinct+pharmacological+dementia+models.">Differential effects of alpha7 nicotinic receptor agonist PHA-543613 on spatial memory performance of rats in two distinct pharmacological dementia models.</a> Behav Brain Res. 278, 404-410 (2015).</li><li>McDonald, J. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Handbook of biological statistics. 2, (2009).</li><li>Castellano, M. A., Diaz-Palarea, M. D., Rodriguez, M., Barroso, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Lateralization+in+male+rats+and+dopaminergic+system:+evidence+of+right-side+population+bias.">Lateralization in male rats and dopaminergic system: evidence of right-side population bias.</a> Physiol Behav. 40, 607-612 (1987).</li><li>Andrade, C., Alwarshetty, M., Sudha, S., Suresh Chandra, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effect+of+innate+direction+bias+on+T-maze+learning+in+rats:+implications+for+research.">Effect of innate direction bias on T-maze learning in rats: implications for research.</a> J Neurosci Methods. 110, 31-35 (2001).</li></ol>

著者が明らかに何もありません。

執事とローリンズのプロトコル31と比較して本研究では、変更が行われました。3 D プリンターは、におけるを作成に使用されました。3 D 印刷は、製品化における手頃な価格でコスト効果の高い選択肢を提供します。ラットの不安を減らすためには、テスト中に、におけるは最低被写体照度が暗い部屋で行われました。ラットが目標の腕の 1 つを入力した後我々 は優しく相?...

アメリカ心臓協会 (2017 年)、心不全 (CA) によると-引き起こされた死亡率 4 分ごとに発生し、米国1年ごとの以上 400,000 人に影響します。CA が不十分な血液灌流2,3,4の結果として神経の脳損傷を引き起こす可能性が定評です。海馬5,6、7学習とメモリ8,9、重要な神経細胞に影響を与えるの虚血に敏感な ca1 で CA による脳損傷が発生します。 10、11,12。また、海馬 (すなわち CA1 ニューロン) の虚血条件下での樹状突起スパイン密度の損失は、空間記憶障害13,14,15で重要な役割を果たしています。CA 後これらの病理学的変化による行動障害など: 不安、うつ病、心的外傷後ストレス障害、記憶喪失がより普及しています。(低体温症) を除く神経保護治療のほとんどが CA16 後機能的転帰の改善に失敗する改良された CA 生存率と相関する医療技術 (すなわち効率的な外来サービス) の進歩をされているが、 ,17。CA の生存者は通常貧しい生活質を持っており、16を支出増分の医療負担。
行動テストで脳梗塞の認知状況の評価は、両方の薬剤の有効性を判断し、最終的に成功した臨床試験を開発することが重要。1940 年代に、エドワード ・ トールマンは、空間記憶の海馬ベース18を勉強する最初の動作試験を設計されています。その後、さまざまな迷路 (モリス水迷路、放射状迷路、T または Y-迷路とバーンズ迷路) は海馬空間学習とラット19,20,21,22 でメモリを評価するため開発されました。 ,23。広く使われている行動テストの 1 つが空間学習を調べ、モリス水迷路とラットのメモリ モデル24。ただし、モリス水迷路では、泳ぐし、完全運動機能とコントロールを発揮するラットが必要です。仮死心不全 (ACA、CA のモデルラット) モデルなど虚血実験、大腿動脈/静脈の穿刺がバイタル血圧、血液ガス、様々 な薬物の導入を取得する必要があります。大腿動脈/静脈穿刺は、ラットの正しく泳ぐ能力をレンダリング脚移動を抑制することが、以来、モリス水迷路は ACA の下で認知障害をテストする最も適切なかもしれない。
バーンズ迷路は、他の広く使用されている行動のテスト齧歯動物モデルにおける記憶・学習能を検査するためです。バーンズ迷路は完全運動機能のコントロール、運動を必要としない、こうしてモリス水迷路よりもストレスの少ない。過去には、我々 はバーンズ迷路を使用して間機能学習・記憶違いが発生するかどうかを確認する実験を行ったコントロールまたは ACA 誘発ラット対偽。データ一方バーンズ迷路に参照 (長期) メモリ25,26を測定に広く用いられてバーンズ迷路という事実のために、軽度中程度の ACA から次認知障害をテストするのには解決していないために,ACA 誘発神経認知赤字より密接に関連作業 (短期) メモリ27,28,29,30バーンズ迷路が私たち ACA の記憶機能を評価するためより少なく実行可能であることを示唆するにはモデル。
従って変更における自発交替テストを使用した ACA の後の作業 (短期) メモリの評価を行った。変更後における自発交替テストの主な利点は簡易性および他の行動のテストと比較してラットに最小限のストレスも重い計算として、事前の動物訓練を必要としない修正におけるという事実のために分析やモリス水迷路とバーンズ迷路に必要なサブルーチン (すなわち、ラットのビデオ イメージ投射)。ここで変更における自発交替テストが十分な解像度を正確に検出し、短期メモリ損失を引き起こす疾患における海馬機能の評価を提供することができます単純なまだ非常に効率的な行動試験パラダイムであることを示す(すなわち ACA)。

<table><tbody><tr><td>3D Printer</td><td>MakerBot</td><td>Replicator</td><td>Fifth generation</td></tr><tr><td>3D Printer Filament</td><td>Hatchbox</td><td></td><td>PLA, 1.75 mm filament diameter</td></tr><tr><td>200 Proof Pure Ethanol</td><td>Koptec</td><td>V1005SG</td><td></td></tr><tr><td>Sani-Chips</td><td>PJ Murphy-Forest Products</td><td></td><td>Size: 8 to 20 mesh; 2.2 cubic foot/package; autoclavable bags</td></tr><tr><td>Rat</td><td>Charles River Laboratories</td><td>Sprague-Dawley</td><td></td></tr><tr><td>Vecuronium bromide</td><td>Sun Pharmaceutical</td><td>47335-931-40</td><td>10 mg</td></tr><tr><td>Epinephrine</td><td>Par Pharmaceutical</td><td>42023-103-01</td><td>Adrenalin Chloride Solution 1 mg/mL, 1:1000</td></tr><tr><td>Buprenorphine Hydrochloride Injection</td><td>Pfizer</td><td>00409-2012-32</td><td>0.3mg/mL</td></tr><tr><td>SketchUp</td><td>Trimble Inc.</td><td></td><td>3D modeling software</td></tr><tr><td>VentElite Small Animal Ventilator</td><td>Harvard Apparatus</td><td>55-7040</td><td>Animals raging in size from mouse to guinea pig (10g to 1kg)</td></tr><tr><td>PowerLab 8/35</td><td>Adinstruments</td><td>PL3508</td><td>8 analog input channels &amp;ndash; 4 of which can be used in differential mode.</td></tr><tr><td>Bio Amps</td><td>Adinstruments</td><td>FE132</td><td>The Bio Amp is a galvanically isolated, high-performance differential bio amplifier optimized for the measurement of a wide variety of biological signals such as ECG, EMG and EEG recordings.</td></tr><tr><td>Quad Bridge Amp</td><td>Adinstruments</td><td>FE224</td><td>A four-channel, non-isolated bridge amplifier designed to allow the PowerLab to connect to most DC bridge transducers.</td></tr><tr><td>LabChart 8</td><td>Adinstruments</td><td></td><td></td></tr><tr><td>ABL80 FLEX CO-OX blood gas analyzer</td><td>Radiometer</td><td></td><td>pH / &lt;em&gt;p&lt;/em&gt; CO&lt;sub&gt;2&lt;/sub&gt; / &lt;em&gt;p&lt;/em&gt; O&lt;sub&gt;2&lt;/sub&gt;</td></tr><tr><td>SURFLO Teflon I.V. Catheter</td><td>Terumo</td><td>sc-361556</td><td>Only use the flexible thin wall catheter (49-mm long)</td></tr><tr><td>Pipet/Infusion Needle</td><td>Hamilton</td><td>7748-03</td><td>17-gauge; 93-mm long; 10-degree angle</td></tr><tr><td>Classic T3 Vaporizer</td><td>SurgiVet</td><td>VCT302</td><td>Classic T3 Isoflurane Funnel Fill</td></tr><tr><td>ENVIRO-PURE Charcoal Canister</td><td>SurgiVet</td><td>32373B10</td><td>Designed to absorb waste anesthetic gas</td></tr><tr><td>O&lt;sub&gt;2&lt;/sub&gt; single flowmeter</td><td>SurgiVet</td><td>32375B1</td><td>0-1000 mL</td></tr><tr><td>N&lt;sub&gt;2&lt;/sub&gt;O Flowmeter</td><td>VetEquip</td><td>401721</td><td>0-4LPM</td></tr><tr><td>Clay Adams Intramedic Luer-Stub Adapter (Sterile)</td><td>Becton Dickinson</td><td>427565</td><td>23 gauge</td></tr><tr><td>Micro Forceps</td><td>Black and Black surgical</td><td>B3FRC-18 RM-8</td><td>7 1/4&quot; (18 cm), 8mm RH, counterweight w/ guide pin 2mm, platform 6 x .3 mm, curved.</td></tr><tr><td>Halstead Mosquito Forceps</td><td>Roboz</td><td>RS-7111</td><td>Curved; 5&quot; Length, 1.3 mm tip diameter, 2.1 mm jaw width</td></tr><tr><td>Mixter Forceps</td><td>Roboz</td><td>RS-7291</td><td>5.25&quot; Curved Extra Delicate, 1.1 mm tips</td></tr><tr><td>Castroviejo Micro Dissecting Spring Scissors</td><td>Roboz</td><td>RS-5650</td><td>Straight, Sharp Points; 9 mm Cutting Edge; 0.15 mm Tip Width; 3 1/2&quot; Overall Length</td></tr><tr><td>Mayo-Stille Scissors</td><td>Roboz</td><td>RS-6891</td><td>5.5&quot; Round Curved</td></tr><tr><td>Dumont #5 Forceps</td><td>Roboz</td><td>RS-5058</td><td>45 Deg Dumoxel Tip Size .10 x .06 mm</td></tr><tr><td>Olsen-Hegar Combination Scissor And Needle Holder</td><td>Roboz</td><td>RS-7884</td><td>Cross Serration Tip; 5.5&quot; Length</td></tr><tr><td>Moloney Forceps</td><td>Roboz</td><td>RS-8254</td><td>Serrated; Slight Curve; 4.5&quot; Length</td></tr></tbody></table>

utilizing the modified t-maze to assess functional memory outcomes after cardiac arrest

背景: 軽度脳虚血 (すなわち心不全) モデルの認知機能障害中等度から評価手術後悪い歩行困難できます。たとえば、手術を受けるとモリス水迷路にさらされるラットできないことがあります、泳ぐこと従って実験を排尿します。
新しい方法: 変更された行動の自発交替におけるテストを設けています。変更におけるプロトコルの主な利点は、虚血後の学習・記憶機能を評価するために十分に強力なその比較的シンプルなデザインです。また、データの分析は、シンプルでわかりやすいです。ラットの学習/記憶障害は軽度から中程度 (6 分) 仮死心不全 (ACA) の有無の両方を決定する、におけるを使いました。ラット探査のための自然な傾向を持っているし、海馬を破壊したラット海馬関連を明らかに減少の自発的交替率の側の好みを採用する傾向にあるに対し、における代替の腕を探索ACA の有無での学習・記憶機能。
結果: ACA グループ側優先率が高くなり、対照と比べて低い交替があります。
既存のメソッドとの比較: のモリス水とバーンズ迷路学習・記憶機能を評価するためより顕著であります。しかし、モリス水迷路は他の迷路よりもストレスです。バーンズ迷路が ACA 誘起 neurocognitive 欠損より密接に関連しています作業 (短期) メモリへ参照 (長期) メモリを測定するために使用されています。
結論: 我々 の全脳虚血モデル (ACA) のワーキング、における経由 (短期) メモリを記述する、シンプルで効果的な戦略を開発しました。

ACA (グローバル脳虚血) は主に作業 (短期) メモリの赤字28,29を発生します。ACA の後、記憶・学習の機能を評価するには、(短期) メモリ30の作業を評価するのに変更された自発交替テストを使用しました。自発交替のテストからの結果は ACA グループ (26.19 ± 4.96%) で 3 日間の連続から交代率がコントロール群 (...

Watch this Scientific Journal Video about Utilizing the Modified T-Maze to Assess Functional Memory Outcomes After Cardiac Arrest at JoVE.com

Utilizing the Modified T-Maze to Assess Functional Memory Outcomes After Cardiac Arrest

心停止後機能メモリの成果を評価するために変更されたにおけるを活用

このプロトコルでは、仮死の心停止による脳虚血における学習・記憶機能を評価する変更におけるの使用について説明します。

Background: Evaluating mild to moderate cognitive impairment in a global cerebral ischemia (i.e. cardiac arrest) model can be difficult due to poor ...

utilizing-modified-t-maze-to-assess-functional-memory-outcomes-after

Research

JoVE Journal

Behavior

12.0K ビュー.  Louisiana State University Health Science Center. このプロトコルでは、仮死の心停止による脳虚血における学習・記憶機能を評価する変更におけるの使用について説明します。

ビデオ: Utilizing the Modified T-Maze to Assess Functional Memory Outcomes After Cardiac Arrest

Use of an Eight-arm Radial Water Maze to Assess Working and Reference Memory Following Neonatal Brain Injury

Working and reference memory are commonly assessed using the land based radial arm maze.&#160;However, this paradigm requires pretraining, food deprivation, and may introduce scent cue confounds. The eight-arm radial water maze is designed to evaluate reference and working memory performance simultaneously by requiring subjects to use extra-maze cues to locate escape platforms and remedies the limitations observed in land based radial arm maze designs. Specifically, subjects are required to avoid the arms previously used for escape during each testing day (working memory) as well as avoid the fixed arms, which never contain escape platforms (reference memory). Re-entries into arms that have already been used for escape during a testing session (and thus the escape platform has been removed) and re-entries into reference memory arms are indicative of working memory deficits. Alternatively, first entries into reference memory arms are indicative of reference memory deficits. We used this maze to compare performance of rats with neonatal brain injury and sham controls following induction of hypoxia-ischemia and show significant deficits in both working and reference memory after eleven days of testing. This protocol could be easily modified to examine many other models of learning impairment.

The eight-arm radial water maze is designed to evaluate reference and working memory performance simultaneously by requiring subjects to use extra-maze cues to locate escape platforms and remedies the limitations observed in land based radial arm maze designs.

新生児脳損傷後の作業とリファレンス記憶を評価〜8アーム放射状水迷路の使用

Working and reference memory are commonly assessed using the land based radial arm maze.&#160;However, this paradigm requires pretraining, food ...

行動学

Quantifying Cognitive Decrements Caused by Cranial Radiotherapy

With the exception of survival, cognitive impairment stemming from the clinical management of cancer is a major factor dictating therapeutic outcome. For many patients afflicted with CNS and non-CNS malignancies, radiotherapy and chemotherapy offer the best options for disease control. These treatments however come at a cost, and nearly all cancer survivors (~11 million in the US alone as of 2006) incur some risk for developing cognitive dysfunction, with the most severe cases found in patients subjected to cranial radiotherapy (~200,000/yr) for the control of primary and metastatic brain tumors1. Particularly problematic are pediatric cases, whose long-term survival plagued with marked cognitive decrements results in significant socioeconomic burdens2. To date, there are still no satisfactory solutions to this significant clinical problem.
We have addressed this serious health concern using transplanted stem cells to combat radiation-induced cognitive decline in athymic rats subjected to cranial irradiation3. Details of the stereotaxic irradiation and the in vitro culturing and transplantation of human neural stem cells (hNSCs) can be found in our companion paper (Acharya et al., JoVE reference). Following irradiation and transplantation surgery, rats are then assessed for changes in cognition, grafted cell survival and expression of differentiation-specific markers 1 and 4-months after irradiation. To critically evaluate the success or failure of any potential intervention designed to ameliorate radiation-induced cognitive sequelae, a rigorous series of quantitative cognitive tasks must be performed. To accomplish this, we subject our animals to a suite of cognitive testing paradigms including novel place recognition, water maze, elevated plus maze and fear conditioning, in order to quantify hippocampal and non-hippocampal learning and memory. We have demonstrated the utility of these tests for quantifying specific types of cognitive decrements in irradiated animals, and used them to show that animals engrafted with hNSCs exhibit significant improvements in cognitive function3.
The cognitive benefits derived from engrafted human stem cells suggest that similar strategies may one day provide much needed clinical recourse to cancer survivors suffering from impaired cognition. Accordingly, we have provided written and visual documentation of the critical steps used in our cognitive testing paradigms to facilitate the translation of our promising results into the clinic.

Cognitive impairment resulting from the radiotherapeutic management of brain tumors represents a clinically intractable condition that adversely impacts quality of life. The capability to critically evaluate potential interventions for ameliorating radiation-induced cognitive decrements ultimately depends on the capability to undertake rigorous quantitative assessments of cognition.

頭蓋放射線療法によって引き起こされた定量認知デクリメント

With the exception of survival, cognitive impairment stemming from the clinical management of cancer is a major factor dictating therapeutic outcome. For ...

医学

The Double-H Maze: A Robust Behavioral Test for Learning and Memory in Rodents

Spatial cognition research in rodents typically employs the use of maze tasks, whose attributes vary from one maze to the next. These tasks vary by their behavioral flexibility and required memory duration, the number of goals and pathways, and also the overall task complexity. A confounding feature in many of these tasks is the lack of control over the strategy employed by the rodents to reach the goal, e.g., allocentric (declarative-like) or egocentric (procedural) based strategies. The double-H maze is a novel water-escape memory task that addresses this issue, by allowing the experimenter to direct the type of strategy learned during the training period. The double-H maze is a transparent device, which consists of a central alleyway with three arms protruding on both sides, along with an escape platform submerged at the extremity of one of these arms.
Rats can be trained using an allocentric strategy by alternating the start position in the maze in an unpredictable manner (see protocol 1; &#167;4.7), thus requiring them to learn the location of the platform based on the available allothetic cues. Alternatively, an egocentric learning strategy (protocol 2; &#167;4.8) can be employed by releasing the rats from the same position during each trial, until they learn the procedural pattern required to reach the goal. This task has been proven to allow for the formation of stable memory traces.
Memory can be probed following the training period in a misleading probe trial, in which the starting position for the rats alternates. Following an egocentric learning paradigm, rats typically resort to an allocentric-based strategy, but only when their initial view on the extra-maze cues differs markedly from their original position. This task is ideally suited to explore the effects of drugs/perturbations on allocentric/egocentric memory performance, as well as the interactions between these two memory systems.

The goal of this protocol is to investigate spatial cognition in rodents. The double-H water maze is a novel test, which is particularly useful to elucidate the different components of learning, consolidation and memory, as well as the interplay of memory systems.

ダブルH迷路：げっ歯類での学習と記憶のための堅牢な行動試験

Spatial cognition research in rodents typically employs the use of maze tasks, whose attributes vary from one maze to the next. These tasks vary by their ...

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 ...

神経科学

Assessing Spatial Memory Impairment in a Mouse Model of Traumatic Brain Injury Using a Radial Water Tread Maze

Despite the recent increase in use of mouse models in scientific research, researchers continue to use cognitive tasks that were originally designed and validated for rat use. The Radial Water Tread (RWT) maze test of spatial memory (designed specifically for mice and requiring no swimming) has been shown previously to successfully distinguish between controlled cortical impact-induced TBI mice and sham controls. Here, a detailed protocol for this task is presented. The RWT maze capitalizes on the natural tendency of mice to avoid open areas in favor of hugging the sides of an apparatus (thigmotaxis). The walls of the maze are lined with nine escape holes placed above the floor of the apparatus, and mice are trained to use visual cues to locate the escape hole that leads out of the maze. The maze is filled with an inch of cold water, sufficient to motivate escape but not deep enough to require that the mouse swim. The acquisition period takes only four training days, with a test of memory retention on day five and a long-term memory test on day 12. The results reported here suggest that the RWT maze is a feasible alternative to rat-validated, swimming-based cognitive tests in the assessment of spatial memory deficits in mouse models of TBI.

Here we present a protocol for a mouse-specific test of cognition that does not require swimming. This test can be used to successfully distinguish controlled cortical impact-induced traumatic brain injury mice from sham controls.

半径方向水迷路迷路を用いた外傷性脳損傷のマウスモデルにおける空間記憶障害の評価

Despite the recent increase in use of mouse models in scientific research, researchers continue to use cognitive tasks that were originally designed and ...

Detecting Behavioral Deficits in Rats After Traumatic Brain Injury

With the increasing incidence of traumatic brain injury (TBI) in both civilian and military populations, TBI is now considered a chronic disease; however, few studies have investigated the long-term effects of injury in rodent models of TBI. Shown here are behavioral measures that are well-established in TBI research for times early after injury, such as two weeks, until two months. Some of these methods have previously been used at later times after injury, up to one year, but by very few laboratories. The methods demonstrated here are a short neurological assessment to test reflexes, a Beam-Balance to test balance, a Beam-Walk to test balance and motor coordination, and a working memory version of the Morris water maze that can be sensitive to deficits in reference memory. Male rats were handled and pre-trained to neurological, balance, and motor coordination tests prior to receiving parasagittal fluid percussion injury (FPI) or sham injury. Rats can be tested on the short neurological assessment (neuroscore), the beam-balance, and the Beam-Walk multiple times, while testing on the water maze can only be done once. This difference is because rats can remember the task, thus confounding the results if repeated testing is attempted in the same animal. When testing from one to three days after injury, significant differences are detected in all three non-cognitive tasks. However, differences in the Beam-Walk task were not detectable at later time points (after 3 months). Deficits were detected at 3 months in the Beam-Balance and at 6 months in the neuroscore. Deficits in working memory were detected out to 12 months after injury, and a deficit in a reference memory first appeared at 12 months. Thus, standard behavioral tests can be useful measures of persistent behavioral deficits after FPI.

The goal of the behavioral tests presented here is to detect functional deficits in rats after traumatic brain injury. Four specific tests are presented that detect deficits in behaviors to reflect the damage to specific brain areas at times extending to one year after injury.

外傷性脳損傷後のラットの行動の赤字を検出

With the increasing incidence of traumatic brain injury (TBI) in both civilian and military populations, TBI is now considered a chronic disease; however, ...

Motor and Hippocampal Dependent Spatial Learning and Reference Memory Assessment in a Transgenic Rat Model of Alzheimer's Disease with Stroke

Alzheimer&#39;s disease (AD) is a debilitating neurodegenerative disease that results in neurodegeneration and memory loss. While age is a major risk factor for AD, stroke has also been implicated as a risk factor and an exacerbating factor. The co-morbidity of stroke and AD results in worsened stroke-related motor control and AD-related cognitive deficits when compared to each condition alone. To model the combined condition of stroke and AD, a novel transgenic rat model of AD, with a mutated form of amyloid precursor protein (a key protein involved in the development of AD) incorporated into its DNA, is given a small unilateral striatal stroke.
For a model with the combination of both stroke and AD, behavioral tests that assess stroke-related motor control, locomotion and AD-related cognitive function must be implemented. The cylinder task involves a cost-efficient, multipurpose apparatus that assesses spontaneous forelimb motor use. In this task, a rat is placed in a cylindrical apparatus, where the rat will spontaneously rear and contact the wall of the cylinder with its forelimbs. These contacts are considered forelimb motor use and quantified during video analysis after testing. Another cost-efficient motor task implemented is the beam-walk task, which assesses forelimb control, hindlimb control and locomotion. This task involves a rat walking across a wooden beam allowing for the assessment of limb motor control through analysis of forelimb slips, hindlimb slips and falls. Assessment of learning and memory is completed with Morris water maze for this behavioral paradigm. The protocol starts with spatial learning, whereby the rat locates a stationary hidden platform. After spatial learning, the platform is removed and both short-term and long-term spatial reference memory is assessed. All three of these tasks are sensitive to behavioral differences and completed within 28 days for this model, making this paradigm time-efficient and cost-efficient.

To investigate the co-morbid Alzheimer&#39;s disease (AD) and stroke condition in a novel model, three behavior tasks are described that assess both motor control and cognitive behaviors. These tasks include the beam-walk task, cylinder task and Morris water maze.

モーターと脳卒中とアルツハイマー病のトランスジェニックラットモデルにおける海馬依存性の空間学習とリファレンスメモリ評価

Alzheimer&#39;s disease (AD) is a debilitating neurodegenerative disease that results in neurodegeneration and memory loss. While age is a major risk ...

A Metric Test for Assessing Spatial Working Memory in Adult Rats Following Traumatic Brain Injury

Impairments to sensory, short-term, and long-term memory are common side effects after traumatic brain injury (TBI). Due to the ethical limitations of human studies, animal models provide suitable alternatives to test treatment methods, and to study the mechanisms and related complications of the condition. Experimental rodent models have historically been the most widely used due to their accessibility, low cost, reproducibility, and validated approaches. A metric test, which tests the ability to recall the placement of two objects at various distances and angles from one another, is a technique to study impairment in spatial working memory (SWM) after TBI. The significant advantages of metric tasks include the possibility of dynamic observation, low cost, reproducibility, relative ease of implementation, and low stress environment. Here, we present a metric test protocol to measure impairment of SWM in adult rats after TBI. This test provides a feasible way to evaluate physiology and pathophysiology of brain function more effectively.

Traumatic brain injury (TBI) is commonly associated with memory impairment. Here, we present a protocol to assess spatial working memory after TBI via a metric task. A metric test is a useful tool to study spatial working memory impairment after TBI.

外傷性脳損傷後の成人ラットの空間ワーキングメモリ評価のための測定基準試験

Impairments to sensory, short-term, and long-term memory are common side effects after traumatic brain injury (TBI). Due to the ethical limitations of ...

A Needle Model for Vascular Cognitive Impairment Studies in Mice

A Mouse Model for Vascular Cognitive Impairment and Dementia Based on Needle-guided Asymmetric Bilateral Common Carotid Artery Stenosis

Vascular cognitive impairment and dementia (VCID) results from vascular brain injury. Given VCID&#39;s high incidence, which is expected to continue rising as the population ages, it is critical to establish a robust animal model for the disease. This paper presents a novel method of creating a mouse model of VCID that is based on asymmetric bilateral common carotid artery stenosis, which mimics human chronic cerebral hypoperfusion caused by carotid atherosclerosis.
Briefly, common carotid arteries (CCAs) are ligated to different gauge needles (32 G for the right CCA and 34 G for the left CCA) using 7-0 silk sutures followed by immediate needle removal. The remaining suture rings cause persistent blood flow reduction and long-term cognitive impairment associated with white matter injury, microinfarcts, and reactive gliosis, thus closely mimicking the pathogenesis of VCID. Importantly, in this needle model, the clinical representations do not revert with time, providing reliable long-term cognitive impairment. Moreover, the survival rate 24 weeks post surgery was 81.6%, which is higher compared to the other established models of VCID with a similar level of blood flow reduction.
Additional advantages include low material cost and compatibility with MRI to monitor brain injury in live animals since no metal is implanted. The main challenge in employing the needle model of VCID is the requirement for developing advanced surgical skills since mouse CCAs are less than 0.6 mm in diameter and are very fragile. High-quality visual representation of the surgery will thus help researchers to master this technique and advance our understanding of VCID, potentially leading to the development of novel therapeutic modalities to decrease the devastating cognitive decline associated with VCID.

The needle method of asymmetric bilateral common carotid artery stenosis is proposed to create a mouse model for vascular cognitive impairment and dementia. It results in longer-term outcomes compared to previously established models and is compatible with live MRI. Visual representation demonstrating the procedure provides guidance for mastering the surgery.

Vascular cognitive impairment and dementia (VCID) results from vascular brain injury. Given VCID&#39;s high incidence, which is expected to continue ...

Anterograde Amnesia

Source: Laboratories of Jonas T. Kaplan and Sarah I. Gimbel&#8212;University of Southern California
Anterograde amnesia is the loss of the ability to form new memories. This can be distinguished from retrograde amnesia, which is the loss of old memories. Anterograde amnesia can result from damage to structures in the brain that are involved in the formation of new memories. Patients who have damage to the structures of the medial temporal lobe, including the hippocampus, amygdala, and the surrounding cortices, often have severe deficits in the formation of certain kinds of memories. These cases can be informative as to how memory is organized in the brain, and how different systems support different kinds of memories. 
In this video, we will test a patient with medial temporal lobe damage on a series of memory tasks designed to distinguish between different forms of memory. First, we will test short-term or working memory, which is the process we use to keep information in mind temporarily. Next, we will test two different forms of long-term memory: explicit and implicit memory. Explicit memories are conscious and easy to verbalize. For example, memories of facts or episodes from our lives are explicit memories. We can easily tell someone what we ate for breakfast, or what city is the capital of France. Implicit memory involves knowledge we gain from experience but that is not easily expressible. For example, knowing how to do things, or becoming habituated to a stimulus are forms of implicit memory. 
These procedures are based in part on studies of the famous patient Henry Molaison, also known by his initials H.M., who had severe anterograde amnesia as a result of a surgery for intractable epilepsy in which parts of both temporal lobes were resected.1 We will perform a test of digit span, which measures short-term memory, a test of paired-associate learning, which measures explicit or declarative memory, and mirror-drawing, a test of implicit skill learning.2

Testing Short- and Long-term Memories in Patients with Brain Injuries

前向性健忘

Source: Laboratories of Jonas T. Kaplan and Sarah I. Gimbel&#8212;University of Southern California
Anterograde amnesia is the loss of the ability to form ...