This study was supported by the American Heart Association (IRG 14330015, PRE 16850005, PRE 20380386, EIA 0740129N), and the National Institutes of Health (RR 2865AR; P20 RR 016477).

以下で説明するすべての手順を見直し、ウェストバージニア大学健康科学センターの施設内動物管理使用委員会によって承認されています。 動物モデルの1選定注：UCMSモデルで最も一般的に使用される種は、スプラーグ・ドーリー、およびWistarラットおよびBALB / cJのマウスを含みます。他のマウスモデルは、限られた有効性を示した（DBA / 2、C57BL / 6）。しかし、環境ストレスへの遺伝的背景と感受性は種間で大きく変動し、研究11,14,15の成果に影響を与える重要な要因であることが示されているように、慎重な検討が、選択した特定のマウスまたはラット系統に与えられるべきです19。 <ol><li>適応年齢の動物を使用して、このプロトコルはLEAに取るように（希望するエンドポイントが17週齢のラットで評価される場合、例えば 、そのラットは生後〜9週でUCMSプロトコルを起動する必要があります）ST 7-9週間完全に抑うつ症状を誘発します。 </li><li> （そうでない場合は、特定のプロトコルによって必要とされる場合を除く）、食品および標準食自由摂取させて、IACUCが承認したケージや設備にUCMSを受けたシングルハウス動物。 </li><li>コントロールはすべての可能な未知の環境ストレスの影響を低減するために、最小限の活性を有する静かな部屋に収容されていることを確認するために特に注意してください。 </li></ol> 2.予測できない慢性的な軽度のストレスプロトコル注：UCMSを受けた動物は、UCMS操作に使用されるクリーンルームに（&gt;週5日）毎日輸送されます。動物は、毎日の下にリストされているストレス要因の1または2にさらされています。ストレスは、無作為化スケジュールで実行され、それぞれの応力は（ 表1を参照）3-4時間の最小値のために投与されます。 <ol><li> 10〜20オンスを注ぐことにより、ベッドを湿らせます。きれいな水の各標準ケージに。の大きさに応じて水の量を調整します完全にベッドを減衰するが、水のプールを生じさせないためにケージ。 3-4時間、湿ったケージ内の動物が存在します。 </li><li>動物を空のケージ（浅風呂、ケージ傾き）を必要とする他のストレッサーに遷移や新鮮な寝具付きの清潔なケージに入れられた後、3-4時間、各ケージから寝具を削除します。 </li><li> 3-4時間のために（寝具なし）は約45°に傾けてケージ。ケージは動物がケージ内を移動する場所に残ります頑丈なオブジェクトを傾斜させることができます。 </li><li> 8時間に及ぶ30分期間の承継に正常な12時間/ 12時間の明/暗サイクルを変更します。後は、通常のサイクルに動物を返します。 </li><li> 3時間のために除去された近隣の動物のケージのそれにそのホームケージから各動物を転送します。 </li><li>各ケージからすべての寝具を取り外し、〜0.25のマウスについてインチ〜約4時間のラットのための0.5インチの深さまで水を加えます。 hypothermを最小限に抑えるため、RT（〜30℃）よりも暖かい水を使用してくださいIAの可能性。きれいなケージに配置前に水曝露の終わりに柔らかいタオルで簡単に説明すると、乾燥動物。 </li><li>捕食者に動物を公開匂い/ 3-4時間ケージに天敵の種から毛皮または尿（10〜20 ml）をランダムに分布サンプル房を追加することで、音が鳴ります。また、うなるまたは3時間ケージに近接した天敵の種からの捕食音を再生します。 </li><li>毎日のストレス期間の終了時に、清潔なケージにすべての動物を配置し、住宅施設に戻します。 </li></ol> 3.動物のモニタリング/グルーミングスコア<ol><li>獣医の治療を必要とするかもしれない傷やただれの発展のために毎日各動物を検査します。軽度の負傷を開発する動物は、獣医師の同意にUCMSプロトコルで継続することができます。 <ol><li>コー​​トグルーミングを評価する際に訓練を受けた二つの研究者がそれぞれの動物を検査し、0-1の規模で毎週コートスコアを割り当てます。このスコアは抑止され個々のスコアを割り当てることにより、採掘された（0〜きれいな1-汚い）8体regions-頭、首、背のコート、腹側のコート、尾、前肢、後肢、および生殖器領域の各へ。同研究者らは、コートのスコアを割り当てる際に一貫性を維持するために、UCMSプロトコル全体でこのタスクを実行することです。 </li><li> （ すなわち 、&gt; 15％）の体重の急激な減少を経験する任意の動物は、獣医スタッフに報告されるべきであり、プロトコルから除去されなければなりません。 </li></ol></li></ol> 4.行動試験と結果の測定注：評価のいくつかの方法は、げっ歯類のUCMSプロトコルからの行動の変化を定量化するために使用することができます。 <ol><li>累積コートグルーミングスコア。 <ol><li>上述したように、毎週コートグルーミングスコアは、各動物に割り当てられます。 </li></ol></li><li>スクローススプラッシュテスト： <ol><li>寝具を防ぐために、綿パッドやタオルを敷いた清潔なケージに動物を置きますそのコートに付着します。各動物の頭と背側のコートに10％ショ糖溶液をスプレーし、5分間の合計グルーミング行動を記録します。 </li><li> （なめたり、傷をつけたりすることによって毛の洗浄として定義される）活性をグルーミングすると周波数（特定の身体部分のグルーミングの回数）レイテンシ（最初のスプレーやグルーミングの開始間のアイドル時間）によって測定されます。 </li></ol></li><li>スクロース嗜好テスト： <ol><li>試験前（w / v）の72時間、1％ショ糖溶液を飲むために動物を慣らすして、2つのボトル（異なるボトル内の水道水に対して1％スクロース溶液）への暴露を提供しています。馴化後、3時間のショ糖液と水道水を自由に摂取アクセスを有する動物を提供しています。 100×[（シ​​ョ糖消費量）/（水消費量+ショ糖消費）]：3時間レコードショ糖液の消費量を、水をタップし、スクロース好みを計算した後。 </li></ol></li><li>尾懸垂試験（マウスのみ）： <ol><li>蘇（尾の先端2 cm）の粘着テープを使用して、安定したロッド（床から20cm）から尾によりマウスを費やしています。 5分以上かけて録音不動時間。彼らは受動的に、完全に動かハングする場合にのみ、マウスを不動と考えられています。 </li></ol></li><li>強制水泳試験： <ol><li>コンテナに入れ動物（マウス〜15〜20センチ、直径;〜ラット55〜60センチ直径）26±0.5℃の温度の水で満たされました。モビリティの最初の試合と5分間の試験期間中に不動に費やした時間の待ち時間を記録します。試験終了時、乾燥するまで吸収紙タオルで覆われて正常な寝具を使用して加熱パッド（30〜35℃）に保持ケージにぬれた動物を配置します。 </li></ol></li><li>高架式十字迷路。 <ol><li>各動物を5分間自由に迷路を探索することができます。レコードの各アームへの全エントリ、および総時間は先験的基準20によれば 、各アームで過ごしました。 </li></ol></li><li> LocomoTORアクティビティ： <ol><li>全体的に動き、細かい動き、動きを飼育し、非アクティブ、急性（30分）の両方のための、および/または慢性（12時間）の期間として定義されるレコードの活性レベル、。カメラの映像を再生し、分析することにより、観察者または事後によってリアルタイムに（一貫性が維持されなければならないが）個々の環境に応じて、ホームケージまたは新しい環境を使用してこれを行います。 </li></ol></li></ol> UCMS議定書の5拡大/バリエーション<ol><li> （早期に開始されている場合）UCMSによって誘発される成果の発展を鈍らするか、または（後半開始されている場合）設立UCMSによって誘発される結果の重大性を逆転させるために彼らの能力を研究するために、適切な治療薬や課題を管理します。これは、個々の研究室やプロトコル11に依存しています。 </li></ol> 6.最終結果アセスメント<ol><li> UCMSプロトコルおよび行動試験の終了後、後続のEXPERの動物を利用特定の仮説、目的や個々の研究室の目標に対処する方法で、形態。 注：例として、これは慢性ストレスに心血管、炎症性または内分泌応答を含むことができます。あるいは、治療的介入は、その有効性と機構的基盤を評価するために、この時点で導入することができます。 </li></ol>

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The authors have nothing to disclose.

Extensive clinical and epidemiological evidence has indicated that chronic stress is one of the most potent precipitating factors for depression. Repeated exposure to stressors may potentiate individual vulnerability to depression and other neuropsychiatric disorders as a consequence of the psychological and physical demands within the body that accumulate over time. The use of validated preclinical animal models offers a valuable translational tool for studying depression and comorbid diseases. Firstly, an ideal animal ...

うつ病の精神疾患は、現在、世界的に障害や病気の負担の主要な原因として認識さである複合体神経障害です。 NIMHは、アメリカ人の約12％が男性1対の影響を受け倍の多くの女性に、うつ病に罹患していると報告しています。米国だけでは、うつ病は、ドルの直接医療費を数十億を占め及び（利益を低下させ、生産性を失った）間接費で推定$ 193 billionより2。うつ病の症状は、無快感症、体重、睡眠サイクルの変化、身体活動や個人衛生の減少、絶望や罪悪感、および/または死または自殺の再発思考が含まれます。この十年間、疫学および臨床研究は、うつ病は、心臓血管疾患（CVD）の罹患率および死亡率3の独立したリスク因子であり、心血管病理のより重度の予後を予測する、を含むことを示していますかかわらず、明白なCVD 4の既往歴のtherosclerosis、高血圧、心筋梗塞、冠動脈疾患、。増加罹患率とうつ病の有害公衆衛生への影響にもかかわらず、この疾患の病因と関連する病態生理学は十分に理解されていないと、様々な要因（例えば、遺伝的、生物学的、および環境要素）に障害の不均一性はに臨床診断が困難になっています定義されています。 証拠は解決できない心理的ストレスがうつ病の病気を開発するための主要な要因であり、また、原因5,6軸視床下部-下垂体-副腎（HPA）の破壊および調節不全に一部、うつ病とCVDを結ぶ強力な病原因子であり得ることを示しています。 HPA軸の機能障害は、うつ病やいくつかのCVD危険因子の開発において観察された行動や生理的な変化に関連している主要なメカニズムであるなど、脂質異常症、肥満、糖尿病7。うつ病の複数の前臨床モデルは、臨床的うつ病の改変されたHPA軸活性特性のメカニズムを再現する試みで開発されてきました。このようなモデルは、動物における慢性及び急性ストレスに関連する行動、神経学的および生理学的変化を調査するための検証手段を提供します。疾患の動物モデルの妥当性は、モデルの設計と解剖学、神経生理学を再現する能力、およびヒト疾患において観察された行動の特徴の病因および進行の関連性に基づいています。また、（などのSSRIなど）の治療に前臨床応答は、臨床現場で観察されたものと同様の結果が得られるはず。 ストレス誘発性うつ病のいくつかの動物モデルは、現在、学習性無力感、早期の生活上のストレス、および社会的敗北ストレスとして、研究に利用されています。しかしながら、これらのモデルの各々は、固有の欠点を有していますそれは、彼らの翻訳効能8を軽減 。過去数十年の中では、予測できない慢性マイルドストレス（UCMS）プロトコルは、げっ歯類9におけるうつ病の病態生理学を研究するための最も翻訳関連モデルの一つとして浮上しています。このモデルは、ストレス要因への慢性曝露はストレス応答システムを破壊し、最終的には抑うつ障害の発症につながる基本的な概念に基づいています。 UCMSプロトコルの間、動物は、日常的に穏やかな環境的・社会的ストレス要因のランダム化された一連のにさらされています。人間の状況に、このモデルの妥当性を向上させる重要な要因は、ストレス要因の予測不可能性と制御不能の高いだけでなく、それらが導入される時間です。さらに、UCMSプロトコルではなく、人生の早い段階または積極的な物理的刺激に頼るので、軽度のストレス因子を使用しています。 UCMS暴露の期間にわたり、抑うつ行動が開発し、CLINに匹敵しますiCalの症状は、報酬（無快感症）、身体活動と調査行動（無力感と絶望）、コート状態の悪化と変更された性的行為10の変化に対する応答性を低下含みます。ほとんどすべての抑うつ症状の実証は、このモデルを使用して報告されており、研究では、これらの行動は、ストレスの停止後数週間持続することが実証されています。密接にヒトでのこれらの薬剤の臨床アクションと変数の有効性を反映治療的改善の類似した神経学的効果を示唆し、また、これらのUCMS誘発性うつ病の行動は徐々に慢性によって尊敬することができますが、特に抗うつ薬ではない急性、治療11,14- 18。ここでは、UCMSプロトコルの詳細な説明を報告し、マウスでの典型的な行動や血管転帰について説明します。

<table border="0" cellpadding="0" cellspacing="0">
	<tbody>
		<tr>
			<td>Temporary animal cages with lids</td>
			<td>n/a</td>
			<td>n/a&#160;</td>
			<td>Provided by your animal care facility</td>
		</tr>
		<tr>
			<td>Rodent bedding</td>
			<td>n/a</td>
			<td>n/a&#160;</td>
			<td>Provided by your animal care facility</td>
		</tr>
		<tr>
			<td>Predator fur or urine</td>
			<td>n/a</td>
			<td>n/a&#160;</td>
			<td>Provided by your animal care facility</td>
		</tr>
		<tr>
			<td>Rodent cage drinking bottles</td>
			<td>n/a</td>
			<td>n/a&#160;</td>
			<td>Provided by your animal care facility</td>
		</tr>
		<tr>
			<td>1000 ml graduated cylinder</td>
			<td>variable</td>
			<td>variable</td>
			<td>This is optional.&#160; Any container from which a known quantiity of water can be poured will be appropriate</td>
		</tr>
		<tr>
			<td>Wooden blocks cut from 2x4</td>
			<td>variable</td>
			<td>variable</td>
			<td>8 inch sections are cut to facilitate cage tilt procedures</td>
		</tr>
		<tr>
			<td>Soft paper towels</td>
			<td>n/a</td>
			<td>n/a&#160;</td>
			<td>Provided by your institution</td>
		</tr>
		<tr>
			<td>Small spray bottle&#160;</td>
			<td>Walmart</td>
			<td>n/a&#160;</td>
			<td>100-200 ml volume is sufficient, used for sucrose splash test</td>
		</tr>
		<tr>
			<td>Medium (mice) or large (rats) plastic tubs for swim testing</td>
			<td>Walmart</td>
			<td>variable</td>
			<td>Should be of sufficient depth that the animals cannot touch bottom (e.g., 2 feet)</td>
		</tr>
	</tbody>
</table>

an unpredictable chronic mild stress protocol for instigating depressive symptoms, behavioral changes and negative health outcomes in rodents

慢性の、未解決の応力が臨床的うつ病の発症の主要な危険因子です。ストレス誘発性うつ病の多くの前臨床モデルが報告されているが、予測不可能なchronic軽度のストレスは（UCMS）は、プロトコルは、無快感症、変更されたグルーミング行動などの一般的に臨床的鬱病に関連する行動症状を誘導するための確立された翻訳関連モデルであり、無力感を学びましたげっ歯類インチUCMSプロトコルはまた、生理的な誘導（E .G。、高コルチゾール血症、高血圧症）および神経学的臨床的うつ病に関連付けられている変更（例えば、無快感症は、無力感を学びました）。重要なことは、UCMS誘発性抑うつ症状は、共通のSSRIと、慢性、急性ではない処理によって改善することができます。このように、UCMSプロトコルは、より極端なストレス要因を利用急性ストレスプロトコルまたはプロトコルよりも多くの利点を提供しています。私たちのプロトコルは、7明確なSTREに無作為化、日常のリスクを伴いますssors：湿った寝具、寝具の除去、ケージチルト、明/暗サイクルの変化、社会的ストレス、浅い水浴、および捕食者は/がにおいが聞こえます。 8週間、3-4時間、毎日これらの軽度のストレス因子にげっ歯類を施すことにより、我々は重要な行動の変化や心血管系への貧しい人々の健康の成果の両方を示しています。このアプローチは、同様に新たな可能性のある治療薬または介入戦略のテストのために、慢性ストレス誘発性のうつ病と関連し、神経学的行動、および生理的な変化の詳細な尋問を可能にします。

UCMS手順次の1週間のスケジュールの例を表1。毎週に示され、ストレッサーは歓呼を防止し、毎日ストレス負荷の予測不可能性を確保するために、無作為化しました。 行動のような抑うつの対策 UCMSの8週間後、彼らの非ストレス対照株に比べて慢性的にストレスを受けげっ歯類の両方の行動や生理的な結果に有意な変化がありました。 UCM...

Watch this Scientific Journal Video about An Unpredictable Chronic Mild Stress Protocol for Instigating Depressive Symptoms, Behavioral Changes and Negative Health Outcomes in Rodents at JoVE.com

An Unpredictable Chronic Mild Stress Protocol for Instigating Depressive Symptoms, Behavioral Changes and Negative Health Outcomes in Rodents

The unpredictable chronic mild stress (UCMS) protocol is a validated method for studying behavioral and physiological changes associated with chronic stress and depressive symptoms. Eight weeks of imposition of the UCMS protocol induces behavioral changes and poor health outcomes in rodents of either gender.

げっ歯類での抑うつ症状を扇動、行動の変化と負の健康アウトカムのための予測不可能な慢性的な軽度のストレスプロトコル

Chronic, unresolved stress is a major risk factor for the development of clinical depression. While many preclinical models of stress-induced depression ...

an-unpredictable-chronic-mild-stress-protocol-for-instigating

Research

JoVE Journal

Behavior

22.2K ビュー.  West Virginia University Health Sciences Center. The unpredictable chronic mild stress (UCMS) protocol is a validated method for studying behavioral and physiological changes associated with chronic stress and depressive symptoms. Eight weeks of imposition of the UCMS protocol induces behavioral changes and poor health outcomes in rodents of either gender.

ビデオ: An Unpredictable Chronic Mild Stress Protocol for Instigating Depressive Symptoms, Behavioral Changes and Negative Health Outcomes in Rodents

Using Chronic Social Stress to Model Postpartum Depression in Lactating Rodents

Exposure to chronic stress is a reliable predictor of depressive disorders, and social stress is a common ethologically relevant stressor in both animals and humans. However, many animal models of depression were developed in males and are not applicable or effective in studies of postpartum females. Recent studies have reported significant effects of chronic social stress during lactation, an ethologically relevant and effective stressor, on maternal behavior, growth, and behavioral neuroendocrinology. This manuscript will describe this chronic social stress paradigm using repeated exposure of a lactating dam to a novel male intruder, and the assessment of the behavioral, physiological, and neuroendocrine effects of this model. Chronic social stress (CSS) is a valuable model for studying the effects of stress on the behavior and physiology of the dam as well as her offspring and future generations. The exposure of pups to CSS can also be used as an early life stress that has long term effects on behavior, physiology, and neuroendocrinology.

This article describes the use of chronic resident intruder social stress as an ethologically relevant paradigm to model postpartum depression and anxiety in lactating rodents.

げっ歯類の授乳でモデル産後うつ病への慢性的社会的ストレスを使用

Exposure to chronic stress is a reliable predictor of depressive disorders, and social stress is a common ethologically relevant stressor in both animals ...

行動学

Nest Building as an Indicator of Health and Welfare in Laboratory Mice

The minimization and alleviation of suffering has moral and scientific implications. In order to mitigate this negative experience one must be able to identify when an animal is actually in distress. Pain, illness, or distress cannot be managed if unrecognized. Evaluation of pain or illness typically involves the measurement of physiologic and behavioral indicators which are either invasive or not suitable for large scale assessment. The observation of nesting behavior shows promise as the basis of a species appropriate cage-side assessment tool for recognizing distress in mice. Here we demonstrate the utility of nest building behavior in laboratory mice as an ethologically relevant indicator of welfare. The methods presented can be successfully used to identify thermal stressors, aggressive cages, sickness, and pain. Observation of nest building behavior in mouse colonies provides a refinement to health and well-being assessment on a day to day basis.

We demonstrate the utility of nest building behavior in laboratory mice as an indicator of welfare. Nest scoring is a sensitive technique that is altered by temperature, illness, and aggression. The time to integrate into nest test (TINT) is a simple cage-side assessment that can detect postoperative pain.

The minimization and alleviation of  suffering has moral and scientific implications. In order to mitigate this  negative experience one must be able to ...

The Forced Swim Test as a Model of Depressive-like Behavior

The goal of the present protocol is to describe the forced swim test (FST), which is one of the most commonly used assays for the study of depressive-like behavior in rodents. The FST is based on the assumption that when placing an animal in a container filled with water, it will first make efforts to escape but eventually will exhibit immobility that may be considered to reflect a measure of behavioral despair. This test has been extensively used because it involves the exposure of the animals to stress, which was shown to have a role in the tendency for major depression. Additionally, the FST has been shown to share some of the factors that are influenced or altered by depression in humans, including changes in food consumption, sleep abnormalities and drug-withdrawal-induced anhedonia. The main advantages of this procedure are that it is relatively easy to perform and that its results are easily and quickly analyzed. Moreover, its sensitivity to a broad range of antidepressant drugs that makes it a suitable screening test is one of the most important features leading to its high predictive validity. Despite its appeal, this model has a number of disadvantages. First, the issue of chronic augmentation is problematic in this test because in real life patients need to be treated for at least several weeks before they experience any relief from their symptoms. Last, due to the aversiveness of the FST, it is important to take into account possible influences it might have on brain structure/function if brain analyses are to be carried out following this procedure.

This protocol describes the forced swim test, which is used for the study of depressive-like behavior in rodents. This procedure involves placing an animal in a container filled with water that eventually will lead to the exhibition of immobility behavior, which is considered to reflect behavioral despair.

うつ病のような動作のモデルとしての強制水泳試験

The goal of the present protocol is to describe the forced swim test (FST), which is one of the most commonly used assays for the study of depressive-like ...

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 ...

A Mouse Model of Subchronic and Mild Social Defeat Stress for Understanding Stress-induced Behavioral and Physiological Deficits

Stressful life events often increase the incidence of depression in humans. To study the mechanisms of depression, the development of animal models of depression is essential. Because there are several types of depression, various animal models are needed for a deeper understanding of the disorder. Previously, a mouse model of subchronic and mild social defeat stress (sCSDS) using a modified chronic social defeat stress (CSDS) paradigm was established. In the paradigm, to reduce physical injuries from aggressors, the duration of physical contact between the aggressor and a subordinate was reduced compared to in the original CSDS paradigm. sCSDS mice showed increased body weight gain, food intake, and water intake during the stress period, and their social behaviors were suppressed after the stress period. In terms of the face validity of the stress-induced overeating and overdrinking following the increased body weight gain, the sCSDS mice may show some features related to atypical depression in humans. Thus, a mouse model of sCSDS may be useful for studying the pathogenic mechanisms underlying depression. This protocol will help establish the sCSDS mouse model, especially for studying the mechanisms underlying stress-induced weight gain and polydipsia- and hyperphagia-like symptoms.

Here, methods for developing a mouse model of subchronic and mild social defeat stress are described and used to investigate the pathogenic features of depression including hyperphagia- and polydipsia-like symptoms following increased body weight.

ストレスによって誘発される行動や生理障害を理解するための亜慢性マイルド社会敗北ストレスのマウスモデル

Stressful life events often increase the incidence of depression in humans. To study the mechanisms of depression, the development of animal models of ...

Feeding Experimentation Device (FED): Construction and Validation of an Open-source Device for Measuring Food Intake in Rodents

Food intake measurements are essential for many research studies. Here, we provide a detailed description of a novel solution for measuring food intake in mice: the Feeding Experimentation Device (FED). FED is an open-source system that was designed to facilitate flexibility in food intake studies. Due to its compact and battery powered design, FED can be placed within standard home cages or other experimental equipment. Food intake measurements can also be synchronized with other equipment in real-time via FED's transistor-transistor logic (TTL) digital output, or in post-acquisition processing as FED timestamps every event with a real-time clock. When in use, a food pellet sits within FED's food well where it is monitored via an infrared beam. When the pellet is removed by the mouse, FED logs the timestamp onto its internal secure digital (SD) card and dispenses another pellet. FED can run for up to 5 days before it is necessary to charge the battery and refill the pellet hopper, minimizing human interference in data collection. Assembly of FED requires minimal engineering background, and off-the-shelf materials and electronics were prioritized in its construction. We also provide scripts for analysis of food intake and meal patterns. Finally, FED is open-source and all design and construction files are online, to facilitate modifications and improvements by other researchers.

Feeding Experimentation Device FED: Construction and Validation of an Open-source Device for Measuring Food Intake in Rodents

Feeding Experimentation Device (FED) is an open-source device for measuring food intake in mice. FED can also synchronize food intake measurements with other techniques via a real-time digital output. Here, we provide a step-by-step tutorial for the construction, validation, and usage of FED.

げっ歯類において食物摂取を測定するためのオープンソースデバイスの構築と検証：実験装置（FED）を給餌

Food intake measurements are essential for many research studies. Here, we provide a detailed description of a novel solution for measuring food intake in ...

An Automated T-maze Based Apparatus and Protocol for Analyzing Delay- and Effort-based Decision Making in  Free Moving Rodents

Many neurological and psychiatric patients demonstrate difficulties and/or deficits in decision making. Rodent models are helpful to produce a deeper understanding of the neurobiological causes underlying the decision-making problems. A cost-benefit based T-maze task is used for measuring decision making in which rodents choose between a high reward arm (HRA) and a low reward arm (LRA). There are two paradigms of the T-maze decision-making task, one in which the cost is a time delay and the other in which it is physical effort. Both paradigms require a tedious and labor-intensive management of experimental animals, multiple doors, pellet reward, and arm choice recordings. In the current work, we invented an apparatus based on traditional T-maze with full automation for pellet delivery, door management and choice recordings. This automated setup can be used for the evaluation of both delay- and effort-based decision making in rodents. With the protocol described here, our lab investigated the decision-making phenotypes of multiple genetically modified mice. In the representative data, we showed that the mice with ablated medial habenular showed aversions of both delay and effort and tended to choose the immediate and effortless reward. This protocol helps to decrease the variability caused by experimenter intervention and to enhance experiment efficiency. In addition, chronic silicon probe or microelectrode recording, fiber-optic imaging and/or manipulation of neural activity can be easily applied during the decision-making task using the setup described here.

This article introduces an automated T-maze apparatus that we invented, and a protocol based on this apparatus for analyzing delay-based decision making and effort-based decision making in free moving rodents.

自動化における遅延および努力ベース無料移動齧歯動物における意思決定を分析するための装置およびプロトコルのベース

Many neurological and psychiatric patients demonstrate difficulties and/or deficits in decision making. Rodent models are helpful to produce a deeper ...

The Unpredictable Chronic Mild Stress Protocol for Inducing Anhedonia in Mice

Depression is a highly prevalent and debilitating condition, only partially addressed by current pharmacotherapies. The lack of response to treatment by many patients prompts the need to develop new therapeutic alternatives and to better understand the etiology of the disorder. Pre-clinical models with translational merits are rudimentary for this task. Here we present a protocol for the unpredictable chronic mild stress (UCMS) method in mice. In this protocol, adolescent mice are chronically exposed to interchanging unpredictable mild stressors. Resembling the pathogenesis of depression in humans, stress exposure during the sensitive period of mice adolescence instigates a depressive-like phenotype evident in adulthood. UCMS can be used for screenings of antidepressants on the variety of depressive-like behaviors and neuromolecular indices. Among the more prominent tests to assess depressive-like behavior in rodents is the sucrose preference test (SPT), which reflects anhedonia (core symptom of depression). The SPT will also be presented in this protocol. The ability of UCMS to induce anhedonia, instigate long-term behavioral deficits and enable reversal of these deficits via chronic (but not acute) treatment with antidepressants strengthens the protocol&#39;s validity compared to other animal protocols for inducing depressive-like behaviors.

Here we present the unpredictable chronic mild stress protocol in mice. This protocol induces a long-term depressive-like phenotype and enables to assess the efficacy of putative antidepressants in reversing the behavioral and neuromolecular depressive-like deficits.

マウスで無快感を誘導するため予測不可能な慢性的な軽度ストレス プロトコル

Depression is a highly prevalent and debilitating condition, only partially addressed by current pharmacotherapies. The lack of response to treatment by ...

Stress-Enhanced Fear Learning, a Robust Rodent Model of Post-Traumatic Stress Disorder

Fear behaviors are important for survival, but disproportionately high levels of fear can increase the vulnerability for developing psychiatric disorders such as post-traumatic stress disorder (PTSD). To understand the biological mechanisms of fear dysregulation in PTSD, it is important to start with a valid animal model of the disorder. This protocol describes the methodology required to conduct stress-enhanced fear learning (SEFL) experiments, a preclinical model of PTSD, in both rats and mice. SEFL was developed to recapitulate critical aspects of PTSD, including long-term sensitization of fear learning caused by an acute stressor. SEFL uses aspects of Pavlovian fear conditioning but produces a distinct and robust sensitized fear response far greater than normal conditional fear responses. The trauma procedure involves placing a rodent in a conditioning chamber and administering 15 unsignaled shocks randomly distributed over 90 minutes (for rat experiments; for mouse experiments, 10 unsignaled shocks randomly distributed over 60 minutes are used). On day 2, rodents are placed in a novel conditioning context where they receive a single shock; then, on day 3 they are placed back in the same context as on day 2 and tested for changes in freezing levels. Rodents that previously received the trauma display enhanced levels of freezing on the test day compared to those that received no shocks on the first day. Thus, with this model, a single highly stressful experience (the trauma) produces extreme fear of the stimuli associated with the traumatic event.

Here we describe the detailed methodology required to conduct stress-enhanced fear learning (SEFL) experiments, a preclinical model of post-traumatic stress disorder, in both rats and mice. The model utilizes aspects of Pavlovian fear conditioning and freezing as an index of enhanced fear in rodents.

恐怖のストレス強化学習、心的外傷後ストレス障害の堅牢な齧歯動物モデル

Fear behaviors are important for survival, but disproportionately high levels of fear can increase the vulnerability for developing psychiatric disorders ...

Novel Object Recognition and Object Location Behavioral Testing in Mice on a Budget

Ethologically relevant behavioral testing is a critical component of any study that uses mouse models to study the cognitive effects of various physiological or pathological changes. The object location task (OLT) and the novel object recognition task (NORT) are two effective behavioral tasks commonly used to reveal the function and relative health of specific brain regions involved in memory. While both of these tests exploit the inherent preference of mice for the novelty to reveal memory for previously encountered objects, the OLT primarily evaluates spatial learning, which relies heavily on hippocampal activity. The NORT, in contrast, evaluates non-spatial learning of object identity, which relies on multiple brain regions. Both tasks require an open-field-testing arena, objects with equivalent intrinsic value to mice, appropriate environmental cues, and video recording equipment and the software. Commercially available systems, while convenient, can be costly. This manuscript details a simple, cost-effective method for building the arenas and setting up the equipment necessary to perform the OLT and NORT. Furthermore, the manuscript describes an efficient testing protocol that incorporates both OLT and NORT and provides typical methods for data acquisition and analysis, as well as representative results. Successful completion of these tests can provide valuable insight into the memory function of various mouse model systems and appraise the underlying neural regions that support these functions.

Here we provide a protocol which includes comprehensive instructions for the economical establishment of murine object location and novel object recognition behavioral testing, including the design, cost, and construction of required equipment as well as execution of behavioral testing, data collection, and analysis.