この作品は、費補助金JSPSフェローのためのAAにすることによってサポートされていました。 ;費補助金のために日本学術振興会科学研究員のための革新的な分野にTKへ（第4501号及び25132712）によりますそして、麻布大学。 図2によって授与研究プロジェクトの助成金によって木原、T.、原田、T.、＆越田、N.ウェハー互換製造およびナノ結晶シリコン熱誘起超音波エミッタの特性から転載されました。 センサおよびアクチュエータA：では、物理 、ボリューム125、エルゼビア、P。 426、（2006）、エルゼビアからの許可を得て。

<html>すべての手順は、麻布大学の倫理委員会によって承認されました。全ての実験は、防音室で行いました。 1.動物の準備<ol><li>記録のための男性<ol><li>交配の経験が性的に成熟した雄マウスを入手します。 </li></ol></li><li>女性被験者<ol><li> （ - 12週齢通常8）ケージあたり2〜5同腹子に収容されてきた処女雌マウスを入手します。 </li><li>マクリーン21によれば、試験前の発情周期の位相を決定するために、毎日の膣スメアを取得します。ケアは、偽妊娠を避けるために、膣の刺激を最小限に抑えるよう注意が必要です。 </li><li>ギムザで塗抹標本を染色し、白血球、角化上皮細胞、および有核上皮細胞の有無に基づいて発情周期の位相を決定し、ネルソン22によります。 </li></ol></li></ol> 2. [デバイス（図1） _content &quot;&gt; <img alt="図1" src="/files/ftp_upload/53074/53074fig1.jpg" /> 二つの選択肢のテストボックスとデバイスの図1の回路図。マウスは、彼らとニュートラルゾーンの間に位置する小さなゲートを介して部屋AとBにアクセスすることができます。二選択ボックスと2つの超音波エミッタは防音室の床に置かれています。マイクは防音チャンバー内に吊り下げられています。略称：PC、パーソナルコンピュータである。 <a href="https://www.jove.com/files/ftp_upload/53074/53074fig1large.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> <ol><li>記録のために<ol><li> USB型A / Dコンバータを使用してPCに超コンデンサーマイクを接続します。 </li><li> 「7」にA / D変換器のゲインレベルを設定します。 </li><li>オーディオキャプチャソフトウェアを使用して、400 kHzでのサンプリングレートで16ビット形式のファイル内の音響データを記録します。 </li></ol></li><li>再生のための<ol><li>使用してPCにD / A変換器を接続しますUSBインターフェース。 </li><li>減衰器を介してD / Aコンバータへの2つのアンプを接続します。 （後者のデバイスは、過負荷を防止するためのものです。） </li><li>各アンプに超音波エミッタを接続します。 </li></ol></li></ol> 3. USV録音<ol><li>側壁で直径6cmの穴0.5センチ金網で覆われた穴小（17×10×11センチ）または中型（21×14×13 cm）のケージにオスのマウスを置きます。中型ケージを使用する場合は、面積を減少させるために、ケージの中央を横切って、パーティションを構成します。 </li><li>録音を汚染する可能性がカサカサ音を回避するためにケージ内のベッドを使用しないでください。 </li><li>メッシュの横にマイクを置きます。 </li><li>ステップ2.1に示す記録セットアップと超音波発声の監視を開始します。 </li><li>雄マウスとケージの中に発情後期または発情間期のいずれかで性的に成熟した雌のマウスを置きます。 </li><li> 5分 - 3のレコードUSVs。 </li></ol> 4. TESTボックス<ol><li>アクリル板（ 図1）を使用して、テストボックスを作成します。 注：ニュートラルゾーン（15×20 cm）であり、部屋AとB（20×10センチ、それぞれ）：試験箱（35×20、および×20センチの高さ）は、3つの区画に分割されています。客室AおよびBは、それぞれ0.5センチメートル金網で覆われている4 cmで径孔を有します。穴やメッシュは中立ゾーンと部屋AとBの間の部屋A、Bの各端作るゲート（5×5センチメートルそれぞれ）に配置されています</li><li>音漏れを防止するためには、各メッシュの周囲のボックスにそれらをシール、ゴム板を使用して、メッシュに面する側を除いて、各超音波エミッタを絶縁します。 </li><li>遮音のために部屋AとBの間の壁の両側にリードシールを置きます。 </li></ol> 5.サウンド再生の準備<ol><li>再生音の作成<ol><li>デジタルオーディオ編集ソフトウェアを使用して記録されたファイルからUSVの20秒のセグメントをキャプチャします。 </li><li>背景なしを選択し、キャプチャ20秒USVセグメントファイルからISE（0.35秒）。雑音区間を繰り返すことにより、20秒のバックグラウンドノイズファイルを作成します。 </li><li>同時に2つのサウ​​ンドファイルを再生するには、「左耳」とのために、他のファイルに1つのファイルで、単一のステレオサウンドファイルとして書き出す」右耳。」 </li><li> 40kHzのカットオフを用いて、高域通過フィルタを使用して音声ファイルをフィルタ。 </li><li>後処理ソフトウェアでノイズリダクションツールを使用してノイズを低減します。選択し、ノイズ低減プロファイルとして何のUSVsを含まないファイルのセグメントをキャプチャし、レベル40とノイズリダクションを実行します。 </li></ol></li><li>再生音のキャリブレーション<ol><li>超音波エミッタとマイクの間のスペースは10cmであることを確認してください。 </li><li>超音波エミッタからの音圧レベルを較正するために、ステップ2.1で説明した構成を用いた超音波モニタリングを行います。デシベル（dB）でマイクで音圧レベルを測定します。 </li><li> USIアッテネータとアンプをngの、ステップ3で記録された男性のUSVsと同じ音圧レベルに超音波エミッタによって生成されたUSVsの音量を調整します。 </li><li> 2つの超音波エミッタとUSVs及び背景雑音からなる音声ファイルを使用する場合、2つの超音波エミッタは、行動試験を行う前にキャリブレーション音（例えば、75 kHzの純音）を使用して、同じ音圧レベルを示すことを確認します。 </li><li> USVsの二つの流れから成るサウンドファイルを再生する場合、テストの前に、同じレベルに生成された2つの音圧レベルを調整します。 </li></ol></li></ol> 6.二選択テスト<ol><li>試験箱のメッシュの後ろに超音波エミッタを配置します。 </li><li>アクリル板付きの客室、A、Bにゲートを閉じ、30分間のニュートラルゾーンに女性対象を慣らします。マウスが逃げることができることを避けるために、アクリル板で箱をカバー。 </li><li> CCを使用して録画を開始しますDカメラは、ボックスの上に取り付けられました。カメラ（ 図1には図示せず）は、全ての3つの区画の面積をカバーします。 </li><li>ゲート及びカバーボードを取り外し、女性が自由に音のゾーンを含む試験箱を探索することができます。 </li><li>女性が両方のメッシュを調査しており、ニュートラルゾーンに戻った後は、5または10分間繰り返し、20秒のサウンドファイルの再生を開始。 </li><li> 5または10分間の行動監視を行っています。 </li><li>前の対象から堆積望ましくない嗅覚を減らすために、70％エタノールを用いた試験の間に試験ケージを清掃してください。 </li><li>ランダム部屋AまたはBに向けて固有の側バイアス効果を避けるために、超音波エミッタAとBの場所を切り替えます</li></ol> 7.統計分析<ol><li>行動イベントスコアのソフトウェアを使用して、次のパラメータを分析：各部屋内のエントリの総数を、各中間ゾーンでの滞在時間の合計。各サウンドゾーンの滞在時間の合計。と超音波エミッタの前に各メッシュの検索の合計時間。 図1は 、「ミドルゾーン」とが何を意味するのかを示して、「サウンドゾーン&quot;。 注意：マウスは、2つのゾーン間の境界線上にとどまる場合には（ 図1、破線は）ヘッドの方向によって位置を決めます。ヘッドメッシュ側を向いている場合には、音声区間での滞在として記録されます。 </li><li>各行動パラメータについては、ウィルコクソンの符号順位検定を使用して部屋AとBの応答を比較し、または0.05の有意水準でのt -testsをペアリング。 </li></ol>

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R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Deficit+in+attachment+behavior+in+mice+lacking+the+mu-opioid+receptor+gene.">Deficit in attachment behavior in mice lacking the mu-opioid receptor gene.</a> Science. 304 (5679), 1983-1986 (2004).</li><li>Hiramoto, 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=Tbx1:+identification+of+a+22q11.2+gene+as+a+risk+factor+for+autism+spectrum+disorder+in+a+mouse.">Tbx1: identification of a 22q11.2 gene as a risk factor for autism spectrum disorder in a mouse.</a> Hum. Mol. Genet. 20 (24), 4775-4785 (2011).</li><li>Ey, E., 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=Absence+of+deficits+in+social+behaviors+and+ultrasonic+vocalizations+in+later+generations+of+mice+lacking+neuroligin4.">Absence of deficits in social behaviors and ultrasonic vocalizations in later generations of mice lacking neuroligin4.</a> Genes Brain Behav. , (2012).</li><li>Roy, S., Watkins, N., Heck, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Comprehensive+analysis+of+ultrasonic+vocalizations+in+a+mouse+model+of+fragile+X+syndrome+reveals+limited,+call+type+specific+deficits.">Comprehensive analysis of ultrasonic vocalizations in a mouse model of fragile X syndrome reveals limited, call type specific deficits.</a> PLoS One. 7 (9), e44816 (2012).</li></ol>

The authors declare that they have no competing interests.

Here, the results of a representative test showed that female mice can discriminate between artificial male USVs and background noise (Figure 3). The conclusion to be drawn from these results is that the discrimination signal is reflected in the duration of stay in the room and sound zone, and in the duration of searching the mesh in the first 5 min of testing, but not in the second 5 min (Figure 3C, E and F). These data indicate that mice become habituated to the playback sounds, possib...

多くの動物は種内の通信のための発声を使用しています。マウスハツカネズミでは、通信信号の一つの重要な種類は、20kHzのより高い周波数を有する超音波発声（USVs）、です。マウスによって放出されたUSVsは男女1-4、メス-メス1、5、およびオス-オス1、6相互作用における社会的認知のコンポーネントと考えられています。彼らは自分の彼女の子犬-検索行動を増加させ、母、ため、仔の生存7から隔離されたときにUSVsも子犬によって放出されます。多くの報告がマウスUSVs 8,9を分析し、分類しているが、受信動物の行動反応と神経機構は少ない10、11を文書化されている。後者はUSVsの諸特性の生物学的意義を明確にする必要があります。これらのメカニズムを明らかにするために、再生実験は、効率的な方法です。最近の再生の研究は、女性のことを明らかにしましたマウスはUSVs 12に引き寄せられ、彼らは彼らの親13、14から異なっている男性からUSVsを好むこと。 この記事では、マウスのUSVの好みを評価するために使用される再生テストを説明します。二選択試験箱を図1に示すように、2つの異なるUSVsは、試験容器の二つの区画に同時に再生することが可能で開発された。試験箱のこのタイプは、三つのサブチャンバ内にテスト領域を分割して音声汚染を防止、鉛の壁を使用。超音波エミッタが各部屋の外に位置しています。部屋や超音波エミッタとの間の壁にはワイヤメッシュで覆われた穴があります。マウスは、3つの部屋の中で自由に移動し、超音波エミッタによって再生USVsに応答するかのように、「メッシュを検索する」の挙動を示すことができます。この試験では、マウスは1音エミッタまたは他の近くに異なる期間の期間にご利用いただけます。これらのパラメータは、敏感measuを得るために記録することができます<html>音の好みの再。 バックUSVsを再生するには、ナノ結晶シリコン熱音響放射体（ すなわち 、「NC-Siのエミッタ」）は、以前の研究15-17のように使用されました。これらのデバイスは、薄膜ヒータ電極、ナノ多孔質シリコン層、及び単結晶シリコンウエハで構成されています。デジタルサウンドファイルは、アナログ信号に変換された後、ヒータ電極を通過します。デバイスは、低歪みと有意な音圧に生じる電圧に依存する熱信号に変換します。この装置は、その中で一意である、機械的な振動に依存する共通の音源とは異なり、振動板を必要とせずに音を再生することができます。エミッタは、20から160キロヘルツ（ 図2）の周波数で平坦な音圧レベルを示し、持続時間、周波数、音圧レベル15、18、19の観点で非常に正確にデジタルで記録されたネズミUSVsを再生することができます。 図3に示す代表的な実験においてve_contentは&quot;&gt;、C57BL / 6（B6）雌のBALB / c（BALB）との間で雄USVsとバックグラウンドノイズを選択させた。また、 図4は、間にB6とBALB女性の選択肢を示していますBALBとB6の男性からの同時USVのプレイバックは、以前の研究で報告されているように14。男性のUSVsの特徴は、B6の間で異なり、BALBは20菌株 。これらの結果により、USVsの魅力が存在するプロトコルを用いて評価することができます示されているように、ここで音は、生きた個体から記録された音響的に分析し、他の個体に再生されます。

<table border="0" cellpadding="0" cellspacing="0">
	<tbody>
		<tr>
			<td>Soundproof chamber</td>
			<td>Muromachi Kikai</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Small cage</td>
			<td>CLEA Japan</td>
			<td>CL-0113-1</td>
			<td></td>
		</tr>
		<tr>
			<td>Middle cage</td>
			<td>CLEA Japan</td>
			<td>CL-0103-1</td>
			<td></td>
		</tr>
		<tr>
			<td>Ultrasound condenser microphones</td>
			<td>Avisoft Bioacoustics</td>
			<td>CM16/CMPA</td>
			<td></td>
		</tr>
		<tr>
			<td>A/D converter</td>
			<td>Avisoft Bioacoustics</td>
			<td>UltraSoundGate116H</td>
			<td></td>
		</tr>
		<tr>
			<td>Audio software</td>
			<td>Avisoft Bioacoustics</td>
			<td>RECORDER USGH</td>
			<td></td>
		</tr>
		<tr>
			<td>Adobe Audition 3.0 / Audio editing software</td>
			<td>Adobe Systems</td>
			<td>Adobe Audition 3.0</td>
			<td></td>
		</tr>
		<tr>
			<td>Nc-Si emitter</td>
			<td>Original</td>
			<td></td>
			<td>not commercially available but it is planned to be so in near future</td>
		</tr>
		<tr>
			<td>D/A converter</td>
			<td>National Instruments</td>
			<td>NI USB-6251 BNC</td>
			<td></td>
		</tr>
		<tr>
			<td>Attenuator</td>
			<td>Original</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Amplifier</td>
			<td>Yamatake</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>PC</td>
			<td>Windows 7 professional</td>
			<td></td>
			<td>Intel&#174; core i7-2600K CPU @ 3.4GHz, 8GB RAM, 64-bit operating system</td>
		</tr>
		<tr>
			<td>Event recorder Excel-macro / Event-scoring software</td>
			<td>original</td>
			<td></td>
			<td>Programmed by Naoto Akagawa &#38; Takeru Yamamoto</td>
		</tr>
		<tr>
			<td>CCD Camera</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Rubber plates (made of elastomer resin)</td>
			<td>Tokyo bouon</td>
			<td>TI-75BK B4</td>
			<td>Cut them to the proper size http://www.piano-bouon.jp/shopping/?pid=1329272401-447630&#38;ca=6&#38;p=3</td>
		</tr>
		<tr>
			<td>Giemsa&#39;s azur eosin methylene blue solution</td>
			<td>Merck Millipore</td>
			<td>1.09204.0500</td>
			<td></td>
		</tr>
	</tbody>
</table>

determining ultrasonic vocalization preferences in mice using a two-choice playback test

Mice emit ultrasonic vocalizations (USVs) during a variety of conditions, such as pup isolation and adult social interactions. These USVs differ with age, sex, condition, and genetic background of the emitting animal. Although many studies have characterized these differences, whether receiver mice can discriminate among objectively different USVs and show preferences for particular sound traits remains to be elucidated. To determine whether mice can discriminate between different characteristics of USVs, a playback experiment was developed recently, in which preference responses of mice to two different USVs could be evaluated in the form of a place preference.
First, USVs from mice were recorded. Then, the recorded USVs were edited, trimmed accordingly, and exported as stereophonic sound files. Next, the USV amplitudes generated by the two ultrasound emitters used in the experiment were adjusted to the same sound pressure level. Nanocrystalline silicon thermo-acoustic emitters were used to play the USVs back. Finally, to investigate the preference of subject mice to selected USVs, pairs of two differing USV signals were played back simultaneously in a two-choice test box. By repeatedly entering a defined zone near an ultrasound emitter and searching the wire mesh in front of the emitter, the mouse reveals its preference for one sound over another. This model allows comparing the attractiveness of the various features of mouse USVs, in various contexts.

USVs は 、図3に示す代表的な実験に聞こえる再生として使用された1 BALB男性（20秒あたり161音節）と同様に、バックグラウンドノイズから記録した。この実験では、7メスのB6マウスは、生後9週で使用しました。サウンドを再生する女性の応答のテストの最高の持続時間を決定するために、行動パラメータは合計10分間の試験時間の最初と最後の5分間別々に分析しまし?...

Watch this Scientific Journal Video about Determining Ultrasonic Vocalization Preferences in Mice using a Two-choice Playback Test at JoVE.com

Determining Ultrasonic Vocalization Preferences in Mice using a Two-choice Playback Test

Ultrasonic vocalizations (USVs) in mice differ depending on age, sex, condition, and genetic background. Using two ultrasound emitters broadcasting simultaneously in different locations, this two-choice test can evaluate murine recognition and preference responses to different characteristics of USVs.

二つの選択肢の再生テストを使用して、マウスにおける超音波発声の設定を決定します

Mice emit ultrasonic vocalizations (USVs) during a variety of conditions, such as pup isolation and adult social interactions. These USVs differ with age, ...

determining-ultrasonic-vocalization-preferences-mice-using-two-choice

Research

JoVE Journal

Behavior

11.0K ビュー.  Azabu University. Ultrasonic vocalizations (USVs) in mice differ depending on age, sex, condition, and genetic background. Using two ultrasound emitters broadcasting simultaneously in different locations, this two-choice test can evaluate murine recognition and preference responses to different characteristics of USVs.

ビデオ: Determining Ultrasonic Vocalization Preferences in Mice using a Two-choice Playback Test

The Successive Alleys Test of Anxiety in Mice and Rats

The plus-maze was derived from the early work of Montgomery. He observed that rats tended to avoid the open arms of a maze, preferring the enclosed ones. Handley, Mithani and File et al. performed the first studies on the plus-maze design we use today, and in 1987 Lister published a design for use with mice. 
Time spent on, and entries into, the open arms are an index of anxiety; the lower these indices, the more anxious the mouse is. Alternatively, a mouse that spends most of its time in the closed arms is classed as anxious. 
One of the problems of the plus-maze is that, while time spent on, and entries into, the open arms is a fairly unambiguous measure of anxiety, time in the central area is more difficult to interpret, although time spent here has been classified as &#x201C;decision making&#x201D;. In many tests central area time is a considerable part of the total test time.
Shepherd et al. produced an ingenious design to eliminate the central area, which they called the &#x201C;zero maze&#x201D;. However, although used by several groups, it has never been as widely adopted as the plus-maze. 
In the present article I describe a modification of the plus-maze design that not only eliminates the central area but also incorporates elements from other anxiety tests, such as the light-dark box and emergence tests. It is a linear series of four alleys, each having increasing anxiogenic properties. It has given similar results to the plus-maze in general. Although it may not be more sensitive than the plus-maze (more data is needed before a firm conclusion can be reached on this point), it provides a useful confirmation of plus-maze results which would be useful when, for example, only a single example of a mutant mouse was available, as, for example, in ENU-based mutagenesis programs.

The plus-maze measures anxiety-like behaviour in rodents. There are two opposite closed and two opposite open arms; anxious rodents avoid the open arms. The central area is neither completely open nor closed, so time spent here is ambiguous and difficult to interpret. Here a modification of the plus-maze protocol eliminating this area is described.

マウスおよびラットにおける不安の連続路地テスト

The plus-maze was derived from the early work of Montgomery. He observed that rats tended to avoid the open arms of a maze, preferring the enclosed ones.  ...

行動学

Contextual and Cued Fear Conditioning Test Using a Video Analyzing System in Mice

The contextual and cued fear conditioning test is one of the behavioral tests that assesses the ability of mice to learn and remember an association between environmental cues and aversive experiences. In this test, mice are placed into a conditioning chamber and are given parings of a conditioned stimulus (an auditory cue) and an aversive unconditioned stimulus (an electric footshock). After a delay time, the mice are exposed to the same conditioning chamber and a differently shaped chamber with presentation of the auditory cue. Freezing behavior during the test is measured as an index of fear memory. To analyze the behavior automatically, we have developed a video analyzing system using the ImageFZ application software program, which is available as a free download at http://www.mouse-phenotype.org/. Here, to show the details of our protocol, we demonstrate our procedure for the contextual and cued fear conditioning test in C57BL/6J mice using the ImageFZ system. In addition, we validated our protocol and the video analyzing system performance by comparing freezing time measured by the ImageFZ system or a photobeam-based computer measurement system with that scored by a human observer. As shown in our representative results, the data obtained by ImageFZ were similar to those analyzed by a human observer, indicating that the behavioral analysis using the ImageFZ system is highly reliable. The present movie article provides detailed information regarding the test procedures and will promote understanding of the experimental situation.

This article presents a protocol for a contextual and cued fear conditioning test using a video analyzing system to assess fear learning and memory in mice.

マウスにおけるビデオ解析システムを用いた文脈と手掛り恐怖条件付け試験

The contextual and cued fear conditioning test is one of the behavioral tests that assesses the ability of mice to learn and remember an association ...

The Modified Hole Board - Measuring Behavior, Cognition and Social Interaction in Mice and Rats

This protocol describes the modified hole board (mHB), which combines features from a traditional hole board and open field and is designed to measure multiple dimensions of unconditioned behavior in small laboratory mammals (e.g., mice, rats, tree shrews and small primates). This paradigm is a valuable alternative for the use of a behavioral test battery, since a broad behavioral spectrum of an animal&#8217;s behavioral profile can be investigated in one single test.
The apparatus consists of a box, representing the &#8216;protected&#8217; area, separated from a group compartment. A board, on which small cylinders are staggered in three lines, is placed in the center of the box, representing the &#8216;unprotected&#8217; area of the set-up. The cognitive abilities of the animals can be measured by baiting some cylinders on the board and measuring the working and reference memory. Other unconditioned behavior, such as activity-related-, anxiety-related- and social behavior, can be observed using this paradigm. Behavioral flexibility and the ability to habituate to a novel environment can additionally be observed by subjecting the animals to multiple trials in the mHB, revealing insight into the animals&#8217; adaptive capacities.
Due to testing order effects in a behavioral test battery, na&#239;ve animals should be used for each individual experiment. By testing multiple behavioral dimensions in a single paradigm and thereby circumventing this issue, the number of experimental animals used is reduced. Furthermore, by avoiding social isolation during testing and without the need to food deprive the animals, the mHB represents a behavioral test system, inducing if any, very low amount of stress.

This protocol describes the modified hole board, which is a behavioral test set-up that comprises the characteristics of an open field and a traditional hole board. This set-up enables the differential analysis of unconditioned behavior of small laboratory mammals as well as the analysis of cognitive abilities.

変更された穴会は - マウスとラットにおける行動、認知と社会的相互作用を測定する

This protocol describes the modified hole board (mHB), which combines features from a traditional hole board and open field and is designed to measure ...

The Social Dimension of Stress: Experimental Manipulations of Social Support and Social Identity in the Trier Social Stress Test

In many situations humans are influenced by the behavior of other people and their relationships with them. For example, in stressful situations supportive behavior of other people as well as positive social relationships can act as powerful resources to cope with stress. In order to study the interplay between these variables, this protocol describes two effective experimental manipulations of social relationships and supportive behavior in the laboratory. In the present article, these two manipulations are implemented in the Trier Social Stress Test (TSST)&#8212;a standard stress induction paradigm in which participants are subjected to a simulated job interview. More precisely, we propose (a) a manipulation of the relationship between different protagonists in the TSST by making a shared social identity salient and (b) a manipulation of the behavior of the TSST-selection committee, which acts either supportively or unsupportively. These two experimental manipulations are designed in a modular fashion and can be applied independently of each other but can also be combined. Moreover, these two manipulations can also be integrated into other stress protocols and into other standardized social interactions such as trust games, negotiation tasks, or other group tasks.

Previous research on the social dimension of stress has focused on two important variables: social identity and social support. This protocol introduces an effective experimental manipulation of these two social variables and describes their implementation in a standard stress induction paradigm (Trier Social Stress Test).

トリーアの社会ストレステストにおけるソーシャルサポートと社会的アイデンティティの実験操作：ストレスの社会的側面

In many situations humans are influenced by the behavior of other people and their relationships with them. For example, in stressful situations ...

Eliciting and Analyzing Male Mouse Ultrasonic Vocalization (USV) Songs

Mice produce ultrasonic vocalizations (USVs) in a variety of social contexts throughout development and adulthood. These USVs are used for mother-pup retrieval1, juvenile interactions2, opposite and same sex interactions3,4,5, and territorial interactions6. For decades, the USVs have been used by investigators as proxies to study neuropsychiatric and developmental or behavioral disorders7,8,9, and more recently to understand mechanisms and evolution of vocal communication among vertebrates10. Within the sexual interactions, adult male mice produce USV songs, which have some features similar to courtship songs of songbirds11. The use of such multisyllabic repertoires can increase potential flexibility and information they carry, as they can be varied in how elements are organized and recombined, namely syntax. In this protocol a reliable method to elicit USV songs from male mice in various social contexts, such as exposure to fresh female urine, anesthetized animals, and estrus females is described. This includes conditions to induce a large amount of syllables from the mice. We reduce recording of ambient noises with inexpensive sound chambers, and present a quantification method to automatically detect, classify and analyze the USVs. The latter includes evaluation of call-rate, vocal repertoire, acoustic parameters, and syntax. Various approaches and insight on using playbacks to study an animal&#39;s preference for specific song types are described. These methods were used to describe acoustic and syntax changes across different contexts in male mice, and song preferences in female mice.

Eliciting and Analyzing Male Mouse Ultrasonic Vocalization USV Songs

Mice produce a complex multisyllabic repertoire of ultrasonic vocalizations (USVs). These USVs are widely used as readouts for neuropsychiatric disorders. This protocol describes some of the practices we learned and developed to consistently induce, collect, and analyze the acoustic features and syntax of mouse songs.

雄マウス超音波発声（USV）曲の抽出と分析

Mice produce ultrasonic vocalizations (USVs) in a variety of social contexts throughout development and adulthood. These USVs are used for mother-pup ...

Psychophysical Tracking Method to Measure Taste Preferences in Children and Adults

The Monell two-series, forced-choice, paired-comparison tracking method provides a reliable measure of sweet taste preferences from childhood to adulthood. The method, which is identical for children, adolescents, and adults, is of short duration (&#60; 15 min), does not rely on sustained attention or place demands on memory (which would yield spurious age differences), and minimizes the impact of language development, making this method amenable to the cognitive limitations of pediatric populations. In this whole-mouth tasting method, subjects are asked to taste (without swallowing) pairs of solutions of different sucrose concentrations and to point to the solution they prefer. Each subsequent pair contains the participant&#39;s preceding preferred concentration and an adjacent stimulus concentration. The procedure continues until the subject chooses either a given concentration of sucrose when paired with both a higher and a lower concentration, or the highest or lowest concentration two consecutive times. Subjects are prevented from reaching response criteria on the basis of first or second position bias by the two-series design of the method, which counterbalances the order of solution presentation within each pair between the series (the weaker concentration is presented first in Series 1, second in Series 2). The geometric mean of the two sucrose concentrations chosen in Series 1 and 2 is an estimate of the participant&#39;s most preferred level of sucrose. Sucrose preference as determined with this laboratory-based measure has been shown to be associated with preference for sugars in foods and beverages and with taste receptor genotype, family history of alcoholism, and race/ethnicity, as well as depressive symptomatology among pediatric populations. The method has real-world relevance and has been applied to determine most preferred level of other tastes (e.g., salt), making it a valuable psychophysical tool.

Sweet taste has powerful hedonic appeal among people of all ages, particularly children. Described herein is a reliable and valid method that can be used to determine the level of sweetness most preferred, making it a valuable psychophysical tool for scientists.

子どもと大人で味の環境を測定するために心理物理追跡方法

The Monell two-series, forced-choice, paired-comparison tracking method provides a reliable measure of sweet taste preferences from childhood to ...

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 ...

Novel Object Recognition Test for the Investigation of Learning and Memory in Mice

The object recognition test (ORT) is a commonly used behavioral assay for the investigation of various aspects of learning and memory in mice. The ORT is fairly simple and can be completed over 3 days: habituation day, training day, and testing day. During training, the mouse is allowed to explore 2 identical objects. On test day, one of the training objects is replaced with a novel object. Because mice have an innate preference for novelty, if the mouse recognizes the familiar object, it will spend most of its time at the novel object. Due to this innate preference, there is no need for positive or negative reinforcement or long training schedules. Additionally, the ORT can also be modified for numerous applications. The retention interval can be shortened to examine short-term memory, or lengthened to probe long-term memory. Pharmacological intervention can be used at various times prior to training, after training, or prior to recall to investigate different phases of learning (i.e., acquisition, early or late consolidation, or recall). Overall, the ORT is a relatively low-stress, efficient test for memory in mice, and is appropriate for the detection of neuropsychological changes following pharmacological, biological, or genetic manipulations.

The object recognition test (ORT) is a simple and efficient assay for evaluating learning and memory in mice. The methodology is described below.

マウスの記憶・学習の調査のための新規オブジェクト認識テスト

The object recognition test (ORT) is a commonly used behavioral assay for the investigation of various aspects of learning and memory in mice. The ORT is ...

Reinstatement of Drug-seeking in Mice Using the Conditioned Place Preference Paradigm

The present protocol describes the Conditioned Place Preference (CPP) as a model of relapse in drug addiction. In this model, animals are first trained to acquire a conditioned place preference in a drug-paired compartment, and after the post-conditioning test, they perform several sessions to extinguish the established preference. The CPP permits the evaluation of the conditioned rewarding effects of drugs related to environmental cues. Then, the extinguished CPP can be robustly reinstated by the non-contingent administration of a priming dose of the drug, and by exposure to stressful stimuli. Both methods will be explained here. When the animal reinitiates the behavioral response, a reinstatement of the conditioned reward is considered to have taken place.
The main advantages of this protocol are that it is non-invasive, inexpensive, and simple with good validity criteria. In addition, it allows the study of different environmental manipulations, such as stress or diet, which can modulate relapse into drug seeking behaviors. However, one limitation is that if the researcher aims to explore the motivation and primary reinforcing effects of the drug, it should be complemented with self-administration procedures, as they involve operant responses of animals.

This protocol describes the Conditioned Place Preference (CPP) as a model of relapse. This procedure permits the measurement of relapse in laboratory animals, considering the impact of drug-associated environmental cues as craving and relapse in abstaining addicts is currently the focus of drug-abuse treatment programs.

エアコンの場所の好みのパラダイムを使用してマウスの薬物探索の復権

The present protocol describes the Conditioned Place Preference (CPP) as a model of relapse in drug addiction. In this model, animals are first trained to ...

Assessment of Social Transmission of Food Preferences Behaviors

Olfactory recognition deficits are suggested to be able to serve as clinical marker to differentiate Alzheimer&#39;s disease (AD) subjects from healthy aging groups. For example, olfactory dysfunction in AD can present as impairment in olfactory recognition, emerging during early stages of the disease and worsening while the disease progresses. The social transmission of food preferences (STFP) task is based on a rudimentary form of communication between rodents concerning distant foods dependent on the transmission of olfactory cues. Healthy wild-type mice would prefer to eat a novel, flavored food that was previously cued by a conspecific, and this food preference would be hampered in transgenic AD mice, such as the APP/PS1 model. Indeed, a strong preference for the cued food in C57Bl6/J mice of 3 months of age was found, and this was reduced in 3 months old transgenic APP/PS1 mice. In summary, STFP task could be a powerful measure to be integrated in present subclinical detection assays of AD.

This paper presents a protocol for the investigation of social transmission of food preference in mice. The advantages and possible applications for this procedure, for instance, in detecting early changes in AD mouse models, are highlighted. To conclude, interpretation of the results in light of critical details are discussed.

食品の好み行動の社会的伝達の評価

Olfactory recognition deficits are suggested to be able to serve as clinical marker to differentiate Alzheimer&#39;s disease (AD) subjects from healthy ...