이 작품은에 의해 지원되었다 그랜트 - 에이드에서 AA에 JSPS 휄로우; 보조금 인 원조 과학 연구에 대한 JSPS 휄로우위한 혁신적인 분야 (제 4501 호 및 제 25132712) TK에에 의해; 와 아자 부 대학. 그림 2에 의해 수여 연구 프로젝트 부여하여 키하라, 토니, 하라다, T., Koshida, N. 웨이퍼 호환 제조 및 나노 결정 실리콘 열 유도 초음파 이미 터의 특성에서 다시 인쇄됩니다. 센서 및 액추에이터 :에서 물리적 볼륨 (125), 엘스 비어, P. (426), (2006), 엘스 비어의 허가.

<html> 모든 절차가 아자 부 대학의 윤리위원회에 의해 승인되었다. 모든 실험은 방음 실에서 실시 하였다. 1. 동물 준비 <ol><li> 녹음을위한 남성 <ol><li> 짝짓기 경험이 성적으로 성숙한 수컷 마우스를 가져옵니다. </li></ol></li><li> 여성 주제 <ol><li> (- 오래된 12 주 보통 8) 케이지 당 2-5 한배 새끼와 함께 보관 된 처녀 암컷 마우스를 가져옵니다. </li><li> 매 클린 (21)에 따라, 검사 전 발정주기의 위상을 결정하기 위해 매일 질 얼룩을 얻습니다. 주의 pseudopregnancy 않도록 질 자극을 최소화하기 위해주의해야한다. </li><li> 넬슨 (22)에있어서, 김사와 얼룩 얼룩 및 존재 또는 백혈구, 각질화 된 상피 세포, 상피 세포 및 핵의 유무에 기초하여 발정주기의 위상을 결정한다. </li></ol></li></ol> 2. 장치 (그림 1) _content &quot;&gt; <img alt="그림 1" src="/files/ftp_upload/53074/53074fig1.jpg" /> 그림 두 선택 테스트 상자 및 장치의 도식. 마우스는 그들과 중립 지역 ​​사이에있는 작은 문을 통해 방 A와 B에 액세스 할 수 있습니다. 두 선택 상자와 두 초음파 방사체는 방음 실의 바닥에 배치된다. 마이크는 방음 실 내부에 일시 중단됩니다. 약기 :. PC, 개인용 컴퓨터 <a href="https://www.jove.com/files/ftp_upload/53074/53074fig1large.jpg" target="_blank">이 그림의 더 큰 버전을 보려면 여기를 클릭하십시오.</a> <ol><li> 기록 <ol><li> USB의 A / D 컨버터를 이용하여 PC에 초음파 콘덴서 마이크로폰을 연결한다. </li><li> &quot;7&quot;에 대한 A / D 컨버터의 이득 레벨을 설정한다. </li><li> 오디오 캡처 소프트웨어를 사용하여 400 kHz의 샘플링 레이트를 갖는 16 비트 포맷 파일의 레코드 데이터 탄성. </li></ol></li><li> 재생 <ol><li> D / 사용하여 PC에 컨버터를 연결USB 인터페이스. </li><li> 감쇠기를 통해 D에 두 앰프 / A 컨버터를 연결합니다. (후자의 디바이스는 과부하를 방지하는 역할을한다.) </li><li> 각 앰프에 초음파 이미 터를 연결합니다. </li></ol></li></ol> 3. USV 기록 <ol><li> 측벽에 6cm 직경의 구멍과 0.5 CM 철망으로 덮여 구멍에 소형 (17 × 10 X 11cm)에 남성 마우스 또는 중간 크기 (21 X 14 X 13cm) 케이지를 놓습니다. 중형 케이지를 사용할 때, 면적을 감소시키는 케이지의 중심을 가로 질러 파티션을 구성. </li><li> 녹음을 오염시킬 수 부스럭 거리는 소음을 방지하기 위해 새장에 침대를 사용하지 마십시오. </li><li> 메시 옆에 마이크를 배치합니다. </li><li> 2.1 단계에 표시된 녹화 설정과 초음파 발성 모니터링을 시작합니다. </li><li> 남성 마우스와 케이지에 metestrus 또는 diestrus 중 하나의 성적으로 성숙한 여성 마우스를 놓습니다. </li><li> 5 분 - 3 녹음 USVs. </li></ol> 4. TEST 박스 <ol><li> 아크릴 보드 (그림 1)를 사용하여 테스트 상자를 구축 할 수 있습니다. 참고 : (× 20 ​​(35), 및 X 20cm 높이)를 세 구획으로 나누어 져 시험 상자 : 중립 지역 (15 X 20cm), 객실 A와 B (20 × 10cm, 각각). 방 및 B는 각각 0.5 ㎝ 철망으로 덮여 4cm 직경 홀을 갖는다. 구멍 메시는 중립 지역 및 객실 A와 B 사이의 객실 A와 B의 각 끝에 확인 게이트 (5 × 5cm 각)에 위치 </li><li> 사운드 누설은 각 메시의 둘레 상자로 밀봉 고무 플레이트를 사용하여, 메쉬에 대향 측면을 제외한 각 초음파 방사기를 절연 방지한다. </li><li> 방음을위한 객실 A와 B 사이의 벽의 양쪽에 배치 리드 씰. </li></ol> 5. 사운드 재생을위한 준비 <ol><li> 재생 사운드의 창조 <ol><li> 디지털 오디오 편집 소프트웨어를 사용하여 기록 된 파일로부터 USV의 20 초 세그먼트를 캡처. </li><li> 선택하지 않고 배경을 캡처이세 20 초 USV 세그먼트 파일에서 (0.35 초). 소음 세그먼트를 반복하여 20 초 배경 잡음 파일을 만듭니다. </li><li> &quot;왼쪽 귀&quot;를 위해 다른 파일을 하나의 파일로, 하나의 스테레오 사운드 파일로 내보낼, 동시에 두 개의 사운드 파일을 재생하려면 &quot;오른쪽 귀.&quot; </li><li> 40 kHz의 컷오프와 같은 고역 통과 필터를 사용하여 사운드 파일을 필터링. </li><li> 후 처리 소프트웨어의 노이즈 감소 도구를 사용하여 소음을 줄일 수 있습니다. 선택하고 노이즈 감소 프로파일과 더 USVs가없는 파일의 세그먼트를 캡처하고, 40 레벨과 노이즈 감소를 실행합니다. </li></ol></li><li> 재생 사운드의 교정 <ol><li> 초음파 이미 터와 마이크 사이의 공간이 10cm 있는지 확인합니다. </li><li> 에미 터에서 초음파의 음압 레벨을 교정하는 단계 2.1에 기재된 구성을 사용하여 초음파 모니터링을 수행한다. 데시벨 마이크 (dB)로 음압 레벨을 측정합니다. </li><li> USI감쇠기 및 증폭기 겨, 3 단계에서 기록한 남성 USVs 동일한 음압 레벨 초음파 에미 터에 의해 발생 USVs의 볼륨을 조정한다. </li><li> 두 초음파 에미 터 및 USVs 및 배경 잡음으로 구성된 사운드 파일을 사용하는 경우, 두 초음파 방출기 행동 시험을 수행하기 전에 보정 사운드 (예를 들어, 75 kHz의 순음)를 사용하여 동일한 음압 레벨을 나타내는 것을 확인한다. </li><li> USVs 두 스트림으로 구성된 사운드 파일을 재생하는 경우, 실험 전에, 동일한 레벨로 생성 된 두 음압 레벨을 조정한다. </li></ol></li></ol> 6. 두 선택 테스트 <ol><li> 시험 상자의 메쉬 뒤에 초음파 이미 터를 놓습니다. </li><li> 아크릴 보드와 함께 객실 A와 B에 문을 닫고 30 분 동안 중립 영역에 여성의 주제를 길들. 마우스를 탈출 할 수있는 방지하기 위해 아크릴 보드 상자를 커버. </li><li> CC를 사용하여 비디오 녹화를 시작합니다D 카메라는 상자 위에 장착. 카메라 (하지 그림 1)은 세 개의 구획의 영역을 다루고 있습니다. </li><li> 게이트와 커버 보드를 제거하고 여성의 사운드 영역을 포함하여 자유롭게 테스트 상자를 탐색 할 수 있습니다. </li><li> 여성 모두 메쉬를 조사하였으며, 중성 영역으로 돌아 오면, 5 또는 10 분 동안 20 초 반복 사운드 파일을 재생 시작한다. </li><li> 5 또는 10 분 동안 행동 모니터링을 실시한다. </li><li> 이전 대상에서 침착 원하지 않는 후각 신호를 감소 70 % 에탄올과 시험 사이의 테스트 새장을 청소합니다. </li><li> 초음파의 이미 터의 위치를​​ 랜덤하게 전환 B 실 또는 B. 향해 고유 측 바이어스 효과를 피하기 </li></ol> 7. 통계 분석 <ol><li> 다음 매개 변수를 분석, 행동 이벤트 득점 소프트웨어를 사용하여 각 방에있는 항목의 총 수를; 각각의 중간 영역에서 체류 전체 기간; 각 사운드 영역에서 체류 전체 기간; 과초음파 방출의 앞에 각 메시의 검색의 총 시간. 그림 1은 &quot;중간 영역&quot;과 무엇을 의미하는지 설명 &quot;사운드 영역을 설정합니다.&quot; 참고 경우 마우스 두 영역 사이의 분할 선 (도 1, 점선)의 머리 방향으로의 위치 결정을 유지한다. 헤드 측 메시에 직면했을 때, 그것은 사운드 존에서 머물 얻었된다. </li><li> 각각의 행동 매개 변수에 대해 0.05의 유의 수준을 갖춘 객실에 대한 반응과 B 윌 콕슨 부호 순위 테스트를 사용하여, 또는 쌍 T의 -tests를 비교합니다. </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=Hearing+loss+associated+with+the+modifier+of+deaf+waddler+(mdfw)+locus+corresponds+with+age-related+hearing+loss+in+12+inbred+strains+of+mice.">Hearing loss associated with the modifier of deaf waddler (mdfw) locus corresponds with age-related hearing loss in 12 inbred strains of mice.</a> Hear. Res. 154 (1-2), 45-53 (2001).</li><li>Haga, S., 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=The+male+mouse+pheromone+ESP1+enhances+female+sexual+receptive+behaviour+through+a+specific+vomeronasal+receptor.">The male mouse pheromone ESP1 enhances female sexual receptive behaviour through a specific vomeronasal receptor.</a> Nature. 466 (7302), 118-122 (2010).</li><li>Grimsley, J. M., Monaghan, J. J., Wenstrup, J. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Development+of+social+vocalizations+in+mice.">Development of social vocalizations in mice.</a> PLoS One. 6 (3), e17460 (2011).</li><li>Sugimoto, 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=A+role+for+strain+differences+in+waveforms+of+ultrasonic+vocalizations+during+male-female+interaction.">A role for strain differences in waveforms of ultrasonic vocalizations during male-female interaction.</a> PLoS ONE. 6 (7), (2011).</li><li>Hanson, J. L., Hurley, L. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Female+presence+and+estrous+state+influence+mouse+ultrasonic+courtship+vocalizations.">Female presence and estrous state influence mouse ultrasonic courtship vocalizations.</a> PLoS One. 7 (7), e40782 (2012).</li><li>Wang, H., Liang, S., Burgdorf, J., Wess, J., Yeomans, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ultrasonic+vocalizations+induced+by+sex+and+amphetamine+in+M2,+M4,+M5+muscarinic+and+D2+dopamine+receptor+knockout+mice.">Ultrasonic vocalizations induced by sex and amphetamine in M2, M4, M5 muscarinic and D2 dopamine receptor knockout mice.</a> PLoS One. 3 (4), (2008).</li><li>Moles, A., Kieffer, B. L., D'Amato, F. 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...

많은 동물 종자 간 통신을위한 발성을 사용합니다. 마우스 뮤스의 musculus에서 통신 신호의 하나의 중요한 유형은 20 kHz의보다 높은 주파수가 초음파 발성 (USVs)이다. 마우스에 의해 방출 USVs는 남성 여성, 1-4 여성 - 여성 1, 5, 남성 - 남성 (1), (6) 상호 작용의 사회 인식의 구성 요소로 간주됩니다. USVs는 그녀의 강아지 - 검색 행동을 증가 어머니로부터 분리되는 경우도 새끼에 의해 방출, 따라서 생존 7 강아지 있습니다. 많은 보고서가 적은 10에 설명 된 분석 마우스 USVs 8, 9, 행동 반응 및 수신 동물의 신경 메커니즘을 분류하고 있지만 11. 후자 USVs의 다양한 특성의 생물학적 의미를 명확하게 할 필요가있다. 이러한 메커니즘을 나타 내기 위해, 재생 실험 효율적인 방법이다. 최근 재생 연구는 여성의 것으로 밝혀졌다마우스는 USVs (12)에 매료되어, 그들은 그들의 부모 (13), (14)에서 다른 남성에서 USVs을 선호. 이 문서에서는 마우스의 USV 환경을 평가하는 데 사용되는 재생 테스트를 설명합니다. 두 선택 테스트 박스가도 1에 도시 된 두 개의 다른 USVs는 테스트 케이스의 두 구획을 동시에 재생할 수있는 개발되었다. 시험 상자의이 유형은 세 개의 서브 - 챔버에서 검사 영역을 분할하여 음성 오염을 방지 납 벽을 사용하여. 초음파 방출은 각 객실 외부에 있습니다. 객실과 초음파 이미 터 사이의 벽에서 철망으로 덮여 구멍이 있습니다. 마우스는 3 방에서 자유롭게 이동하고, 초음파 에미 터에 의해 재생 USVs에 응답하는 것처럼, &quot;메시 검색&quot;동작을 보여줄 수있다. 이 시험에서, 생쥐는 하나의 사운드 이미 터 또는 서로 근접 상이한 지속 기간까지. 이러한 매개 변수는 민감한 measu를 얻을 기록 할 수 있습니다<html> 사운드 환경 설정의 재. 다시 USVs를 재생하려면, 나노 결정 실리콘 열 음향 방출 (즉, &quot;NC-시 터&quot;)는 이전의 연구 15-17에서 사용 하였다. 이러한 장치는 박막 가열기 전극, 나노 다공성 실리콘 층과 단결정 실리콘 웨이퍼로 구성된다. 디지털 사운드 파일을 아날로그 신호로 변환 한 후 히터 전극을 통과한다. 장치는 낮은 왜곡과 상당한 음압으로 인한 전위 의존성 열 신호를 변환한다. 기계적 진동에 의존하는 일반적인 소리 발생기,이 진동판의 필요없이 음향을 재생 달리이 장치는 점에서 독특하다. 이미 터 (20)로부터 160 kHz에서 (도 2)까지의 주파수에서 평면 음압 레벨을 나타내고, 그리고 시간, 주파수 및 음압 레벨 15, 18, ​​19의 측면에서 매우 정확하게 디지털 기록 뮤린 USVs를 재생할 수있다. 도 3에 도시 된 대표적인 실험에서 ve_content은 &quot;&gt;, C57BL / 6 (B6) 암컷 BALB / C (BALB) 사이 남성 USVs 및 배경 잡음을 선택하는 것이 허용되었다. 또한,도 4는 사이 B6 및 BALB 암컷의 선택을 도시 BALB 및 B6 남성에서 동시 USV의 플레이는 이전의 연구에서보고 된 14. 남성 USVs의 특성은 B6 사이의 차이와 BALB 20 종자. 이러한 결과에 의해, USVs의 매력은 본 프로토콜로 평가 될 수있다 바와 같이, 하는 소리, 라이브 개인으로부터 기록 된 음향 분석 및 기타 개인 재생됩니다.

<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.

USVs는도 3에 도시 된 대표적인 실험에서 사운드 재생으로 사용 하였다 한 BALB-남성 (20 초 당 161 음절)뿐만 아니라 배경 잡음에서 기록 하였다.이 실험에서, 7 여성 B6 마우스는 9 주령에 사용 하였다 . 사운드를 재생하기 위해 여성의 응답 테스트의 가장 시간을 결정하기 위해, 행동 파라미터는 총 10 분의 시험 시간의 처음과 마지막 다섯 분 동안 별도로 분석 하였다. <p class="jove_content"...

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.

두 선택 재생 테스트를 사용하여 마우스의 초음파 발성 환경 설정을 결정

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 조회수.  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.

비디오: 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.

마우스 및 쥐에있는 불안의 연속 골목길 테스트

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.  ...

연구

행동분석학

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.

마우스에서 비디오 분석 시스템을 사용하여 콘텐츠 및 시작됨 공포 컨디셔닝 테스트

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.

수정 된 구멍 보드 - 마우스 및 쥐의 행동,인지 및 사회적 상호 작용을 측정

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).

스트레스의 사회적 차원 : 트리어 사회에서 사회적지지와 사회적 정체성의 실험 노는 시험 스트레스

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.

수컷 마우스 초음파 발성 (USV) 노래 추출 및 분석

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.

정신 물리학 추적 방법은 어린이와 성인의 맛의 기본 설정을 측정하는

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.

설치류 심리 운동 경계 시험 (rPVT) : 쥐에서 신경 행동 학적 성능을 평가하는 방법

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.

학습 및 생쥐에서 메모리의 조사에 대 한 새로운 개체 인식 테스트

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.

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.

식품 환경 설정 행동의 사회적인 전송에 대 한 평가

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