이 작품은 베일러 대학 연​​구위원회의 승인에 의해 간질 재단의 연구 보조금 지원됩니다.

다음 실험에 사용 된 생쥐는 14 시간 빛과 10 시간의 어둠 (20시부터 6시까지 시간) 낮의주기와, 22 ° C의 주변 온도에서 생성 베일러 대학에서 지내게되었다. 쥐들이 음식과 물에 임의로 액세스를 부여했다. 마우스에 대한 모든 절차는 실험 동물의 관리 및 사용을위한 건강 지침의 국립 연구소에 부합했고, 동물의 프로토콜은 베일러 대학의 동물 관리 및 사용위원회에 의해 승인되었습니다. 개요 추적 에어컨 공포의 작업은 Wiltgen 및 동료 (9)에 의해 설명 된 절차에 근거한다. 1. 장비의 제조 공포 조절 장치 실 (26 센티미터 X 22cm X 18cm)은 아크릴 두 가지 측면, 금속 두 측면과 가벼운 발 충격을 전달하는 데 사용되는 격자 마루 바닥으로 구성되어 있습니다. 시험 챔버는 집소리 감쇠 챔버 D. 챔버는 또한 동작 감지 소프트웨어에 영향을 미치는 외부에서 빛을 방지하기 위해 빛을 꽉. <ol><li> 테스팅 챔버에 대한 충격 레벨, 광 레벨 및 소리 세기 레벨을 보정. 테스트 챔버의 배경 수준을 측정한다. 이 챔버의 배경 소음은 65dB이다. 이 수준을 측정하는 사​​운드 미터를 사용합니다. 미터는 70 dB로 설정 느린 설정 C에, 검출로 설정해야합니다. <ol><li> 0.5 mA에 충격 수준을 보정합니다. 제대로 충격 레벨 ​​(재료 표 참조)을 보정하기 위해 외부 교정 장치를 사용하십시오. 충격 발생기의 내부 둘레는 정확하지 않다. 충격 발생기 정확하게 표준 A 측정기에 의해 측정 할 수없는 스크램블링 충격을 관리한다. </li><li> 그리드 바의 하나 하나에 리드를 배치하고 다른 리드 3 또는 4 개의 바 위에 놓습니다. 충격을 관리하는 충격 생성기를 사용합니다. 올바른 수준이 달성 될 때까지 충격 레벨을 조정합니다. 개에 대해이 작업을 수행채널 조절 챔버를 우려하고있다. </li><li> 실의 조명 수준을 측정 할 때 실에 문을 닫습니다. 약 1.0 광량을 보정합니다. 이 실험에 사용 된 FreezeFrame 소프트웨어에 고유 번호입니다. 외부 빛 미터는 2 룩스로이 글을 읽을 것입니다. 광 레벨은 광 집의 위치를​​ 이동시킴으로써 또는 렌즈의 응축기를 조정함으로써 조정될 수있다. 조정 후 렌즈의 조정 나사를 조여 있는지 확인합니다. </li><li> 85dB에 소리의 데시벨 수준을 보정합니다. 데시벨 (재료 표 참조) 교정 시험 케이지 내부에 외부 데시벨 측정기를 사용합니다. 제시 소리는 2,700 Hz 톤이 될 것입니다. 참고 : 6 개월 이전의 마우스를 사용하는 경우는 옛 마우스가 적자를 듣고 한 수 있기 때문에 백색 잡음을 사용하는 것이 좋습니다 수 있습니다. </li></ol></li><li> 장치는 별도의 보관실에 과목을 준비 후. 참고 : 테스트 룸 같은 방에 마우스를 수용하지 마십시오. </li><li> 레이블 일테스트를 위해 테스트됩니다 생쥐의 전자 꼬리. 이 테스트를하기 전에 과도한 취급을 감소하는 것이 가장 좋습니다. 선택적으로, 꼬리는 실험 전날 스트레스를 줄이기 위해 처리 표지 될 수있다. 마우스는 그 30 분 동안 방에 적응 할 수 있도록 표시 한 후. 테스트를 완료 한 후 마우스를 주택에 대한 별도의 깨끗한 새장이있다. </li></ol> 2. 컨디셔닝 1 일 추적 <ol><li> 케이지에서 각 마우스를 제거하고 공포 조건화 챔버에 전송에 대한 개별 케이지에 넣습니다. 각 케이지 깨끗한 침구를 사용합니다. 마우스는 정확한 테스트의 순서를 유지하기 위해 전송 케이지에 스티커 메모를 배치합니다. 참고 : 마우스가 단독으로 보관하는 경우, 그들은 자신의 홈 케이지에 수송 될 수있다. </li><li> 시험 챔버에 마우스를 놓고 문을 닫습니다. 소프트웨어 프로그램을 시작합니다. </li><li> 훈련 날에, 마우스는 3 분 동안 챔버를 탐험 할 수 있습니다. 소프트웨어는 20 초 톤 (85dB, 2,700 Hz에서) t을 제공합니다동물 오. 20 초의 추적 기간 후 가벼운 충격 (2 초, 0.5 mA)은 동물에게 투여된다. </li><li> 그들은 비디오를보고 혐오 자극을 받았다는 것을 확인하는 피사체의 반응을 기록한다. 200 초 간 시험 간격은 5 조절 시험을 분리한다. 각 시험은 20 초 지연 충격 뒤에 20 초 톤으로 구성되어 있습니다. </li><li> 시험은 동물 시험 케이지에서 분리하기 전에 1 분 동안 테스팅 챔버에 남아 있도록 완료된 후. </li><li> 다시 전송 케이지에 동물을 배치하고 자신의 홈 케이지에 반환. 추가 생쥐는 자신의 홈 케이지에있는 경우 개별적 모두 마우스 테스트를 완료 할 때까지 마우스를 집. 이 테스트되지 않은 다른 쥐에 스트레스를 줄일 수 있습니다. 대체 솔루션은 단독으로 지속적으로 하나의 케이지에서 마우스를 제거의 영향을 줄이기 위해 시험 전에 1 주일 마우스를 모두 수용하는 것입니다. </li><li> EAC 후 30 % 이소프로판올로 테스팅 챔버 청소H 동물은 시험한다. </li><li> 반복 테스트 일대의 모든 마우스에 대한 2.2-2.7 단계를 반복합니다. </li><li> 일대의 마지막 마우스를 테스트 한 후 그들의 식민지 룸으로 마우스를 모두 반환합니다. </li></ol> 3. 추적 공포 컨디셔닝 주 2 : 트레이스 메모리 테스트 <ol><li> 추적 메모리 테스트는 2 일에 발생합니다. 이 프로토콜에서 3 톤의 프리젠 테이션이 있습니다. 추적 컨디셔닝 테스트를위한 새로운 맥락에서 마우스를 놓습니다. </li><li> 세 20 초 톤 프리젠 테이션 다음에 2 분 기준 기간으로 프로그램을 실행할 수있는 소프트웨어를 준비합니다. 각 톤의 프리젠 테이션 사이의 SA 220 초 ITI 있습니다. </li><li> 새로운 상황 조건에 대해, 형상, 조직, 및 조절 챔버의 색깔을 변경하기 위해 상기 챔버의 바닥에 투명 아크릴 인서트를 배치. </li><li> 바닥에서 삽입 무게 보트에 바닐라 추출액을 배치하여 챔버의 냄새를 변경합니다. </li><li> 70 % 에탄올 대신에 30 % 이소프로판올로 챔버를 청소합니다. 참고 :이 도움이 될 것입니다새로운 컨텍스트를 만들 수 있습니다. </li><li> 대기실에 마우스를 가져오고, 필요한 경우 테스트를 위해 자신의 꼬리에 레이블을 재 지정. </li><li> 갈가리 찢긴 종이로 침구를 교체하여 새로운 컨텍스트 전송 새장을 준비한다. 참고 :이 새로운 컨텍스트를 만드는 데 도움이 될 것입니다. </li><li> 다음 프로그램을 시작 테스트 챔버에서 쥐를 놓습니다. 테스트를 완료 한 후 70 % 에탄올로 챔버를 청소합니다. </li><li> 추적 조절이 완료된 후 자신의 홈 케이지에 마우스를 반환합니다. 모든 쥐가 테스트 한 때 그들의 식민지 룸으로 마우스를 모두 반환합니다. </li></ol> 4. 추적 공포 컨디셔닝 3 일 : 문맥 메모리 테스트 <ol><li> 셋째 날 상황에 맞는 조절을 실시한다. 13 분 동안 냉동 동작을 기록하는 프로그램을 실행하는 소프트웨어를 준비한다. </li><li> 테스트하기 전에 각 마우스를 테스트 한 후 30 % 이소프로판올로 챔버를 청소합니다. 문맥은 하루에 1과 동일해야합니다. 전송 케이지는 하루 1과 동일해야합니다. </li><li&gt; 대기실에 마우스를 가져오고 테스트를 위해 자신의 꼬리를 라벨. </li><li> 다음 프로그램을 시작 테스트 실에서 각 마우스를 놓습니다. 테스트를 완료 한 후 30 % 이소프로판올로 챔버를 청소한다. </li><li> 완료되었을 때 그들의 식민지 룸으로 마우스를 모두 반환합니다. </li></ol>

<ol>
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U.S.A. 99, 8980-8985 (2002).</li><li>Crestani, F., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Decreased+GABAA-receptor+clustering+results+in+enhanced+anxiety+and+a+bias+for+threat+cues.">Decreased GABAA-receptor clustering results in enhanced anxiety and a bias for threat cues.</a> Nat. Neurosci. 2, 833-839 (1999).</li><li>Moore, M. D., 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=Trace+and+contextual+fear+conditioning+is+enhanced+in+mice+lacking+the+alpha4+subunit+of+the+GABA(A)+receptor.">Trace and contextual fear conditioning is enhanced in mice lacking the alpha4 subunit of the GABA(A) receptor.</a> Neurobiol. Learn. Mem. 93, 383-387 (2010).</li><li>Cushman, J. D., Moore, M. D., Jacobs, N. S., Olsen, R. W., Fanselow, M. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Behavioral+pharmacogenetic+analysis+on+the+role+of+the+alpha4+GABA(A)+receptor+subunit+in+the+ethanol-mediated+impairment+of+hippocampus-dependent+contextual+learning.">Behavioral pharmacogenetic analysis on the role of the alpha4 GABA(A) receptor subunit in the ethanol-mediated impairment of hippocampus-dependent contextual learning.</a> Alcohol Clin. Exp. Res. 35, 1948-1959 (2011).</li><li>Raybuck, J. D., Lattal, K. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Double+dissociation+of+amygdala+and+hippocampal+contributions+to+trace+and+delay+fear+conditioning.">Double dissociation of amygdala and hippocampal contributions to trace and delay fear conditioning.</a> PLoS ONE. 6, (2011).</li><li>Kwapis, J. L., Jarome, T. J., Schiff, J. C., Helmstetter, F. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Memory+consolidation+in+both+trace+and+delay+fear+conditioning+is+disrupted+by+intra-amygdala+infusion+of+the+protein+synthesis+inhibitor+anisomycin.">Memory consolidation in both trace and delay fear conditioning is disrupted by intra-amygdala infusion of the protein synthesis inhibitor anisomycin.</a> Learn. Mem. 18, 728-732 (2011).</li><li>Gilmartin, M. R., Kwapis, J. L., Helmstetter, F. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Trace+and+contextual+fear+conditioning+are+impaired+following+unilateral+microinjection+of+muscimol+in+the+ventral+hippocampus+or+amygdala,+but+not+the+medial+prefrontal+cortex.">Trace and contextual fear conditioning are impaired following unilateral microinjection of muscimol in the ventral hippocampus or amygdala, but not the medial prefrontal cortex.</a> Neurobiol. Learn. Mem. 97, 452-464 (2012).</li><li>Baysinger, A. N., Kent, B. A., Brown, T. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Muscarinic+receptors+in+amygdala+control+trace+fear+conditioning.">Muscarinic receptors in amygdala control trace fear conditioning.</a> PLoS ONE. 7, (2012).</li><li>Wanisch, K., Tang, J., Mederer, A., Wotjak, C. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Trace+fear+conditioning+depends+on+NMDA+receptor+activation+and+protein+synthesis+within+the+dorsal+hippocampus+of+mice.">Trace fear conditioning depends on NMDA receptor activation and protein synthesis within the dorsal hippocampus of mice.</a> Behav. Brain. 157, 63-69 (2005).</li><li>Smith, D. R., Gallagher, M., Stanton, M. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Genetic+background+differences+and+nonassociative+effects+in+mouse+trace+fear+conditioning.">Genetic background differences and nonassociative effects in mouse trace fear conditioning.</a> Learn. Mem. 14, 597-605 (2007).</li><li>Rudy, J. W., O'Reilly, R. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Contextual+fear+conditioning,+conjunctive+representations,+pattern+completion,+and+the+hippocampus.">Contextual fear conditioning, conjunctive representations, pattern completion, and the hippocampus.</a> Behav. Neurosci. 113, 867-880 (1999).</li><li>Wiltgen, B. J., Sanders, M. J., Anagnostaras, S. G., Sage, J. R., Fanselow, M. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Context+fear+learning+in+the+absence+of+the+hippocampus.">Context fear learning in the absence of the hippocampus.</a> J. Neurosci. 26, 5484-5491 (2006).</li><li>Reijmers, L. G., Perkins, B. L., Matsuo, N., Mayford, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Localization+of+a+stable+neural+correlate+of+associative+memory.">Localization of a stable neural correlate of associative memory.</a> Science. 317, 1230-1233 (2007).</li><li>Huerta, P. T., Sun, L. D., Wilson, M. A., Tonegawa, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Formation+of+temporal+memory+requires+NMDA+receptors+within+CA1+pyramidal+neurons.">Formation of temporal memory requires NMDA receptors within CA1 pyramidal neurons.</a> Neuron. 25, 473-480 (2000).</li><li>Jacobs, N. S., Cushman, J. D., Fanselow, M. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+accurate+measurement+of+fear+memory+in+Pavlovian+conditioning:+Resolving+the+baseline+issue.">The accurate measurement of fear memory in Pavlovian conditioning: Resolving the baseline issue.</a> J. Neurosci. Methods. 190, 235-239 (2010).</li></ol>

저자는 공개 아무것도 없어.

추적 공포 조건화의 기초가되는 신경 회로를 규명 한 여러 연구가 있었다. 추적 공포 조건화는 해마 12-14의 CA1을 포함하는 것으로 생각됩니다. 증거는 내측 전두엽 피질 (mPFC가) 추적 눈 깜짝 에어컨 (15)에 큰 역할을하고 mPFC가 추적 공포 조건화에 관여하는 것으로 발견되었다고도 있습니다. 한 연구는 따라서 추적 간격을 17 동안 메모리를 유지 할 수있는 구조를 제공, mPFC ?...

공포 조건화에서 중립 자극 (NS)을 혐오 무조건 자극 (미국)와 결합된다. NS는 일반적으로 톤이며, 미국과 반복 짝을 조건 자극 (CS)가됩니다. CS는 혐오 미국의 부재, 예컨대 냉동 같은 조건 반응 (CR)을 유도 할 수있다. 일반적으로 사용되는 공포 조건화 프로토콜은 지연 조절입니다. 이 프로토콜에서 NS와 미국의 발병은 연속 또는 자극 프레 젠 테이션에서 일부 중복이되고 있습니다. 동시 에어컨, 이전 버전과의 조화, 그리고 추적 에어컨 1 : 지연 공포 조건화가 시간적 연관 조절의 가장 일반적으로 사용되는 유형 중 하나이지만, 연관 조절 시간적 배열의 몇 가지 다른 종류가 있습니다. 추적 공포 조건화에서 &quot;추적&quot;기간의 결과로 몇 초 NS과 미국 사이의 자극이없는 간격이있다. 몇몇 연구는 적자를보고있다추적 공포 조건화에서의 신경 독성 병변이 구조에서 생산되는 때 해마 2-5 또는 약물 에이전트가 해마 수용체의 기능을 차단하는 데 사용됩니다에 입력. 추적 에어컨 및 상황에 맞는 조절에 적자에서 해마의 결과 병변하지만, 지연의 공포 조건화 8을 손상하지 않습니다. 추적 공포 조건화를 사용하면 여러 가지 이점을 얻을가 있습니다. 공포 조건화 프로토콜은 세 가지 일의 테스트 기간 동안 달성 공간적으로 의존하지 해마에 의존하는 기억을 허용 할 수 있습니다. 추적 공포 조건화는 모리스 물 미로, 새로운 물체 인식 시험, 또는 해마에 의존하는 메모리를 조사하는 다른 미로 테스트를 보완하는 검사로 사용할 수 있습니다.

<table border="0" cellpadding="0" cellspacing="0" width="741">
	<colgroup>
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		<col />
		<col />
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	</colgroup>
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		<tr height="22">
			<td height="22" style="height:22px;width:216px;">FreezeFrame</td>
			<td style="width:143px;">Coulbourn</td>
			<td style="width:119px;"></td>
			<td style="width:263px;"></td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;width:216px;">30% Isopropanol</td>
			<td style="width:143px;"></td>
			<td style="width:119px;"></td>
			<td style="width:263px;">Purchase 90% isopropanol and dilute it down to 30%</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">70% Ethanol</td>
			<td style="width:143px;"></td>
			<td style="width:119px;"></td>
			<td style="width:263px;"></td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;width:216px;">Amp-meter</td>
			<td style="width:143px;">Med-Associates</td>
			<td style="width:119px;">ENV-420</td>
			<td style="width:263px;">Windows XP, Vista, and 7 Compatible (32-bit only)</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Digital Sound Level Meter</td>
			<td style="width:143px;"></td>
			<td style="width:119px;">33-2055</td>
			<td style="width:263px;"></td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;width:216px;">Vanilla Extract</td>
			<td style="width:143px;">McCormick Pure Vanilla Extract</td>
			<td style="width:119px;"></td>
			<td style="width:263px;"></td>
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			<td height="21" style="height:21px;width:216px;">Sticky Notes</td>
			<td style="width:143px;">Post-it</td>
			<td style="width:119px;"></td>
			<td style="width:263px;">3X3 inch</td>
		</tr>
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</table>

trace fear conditioning in mice

이 실험에서 우리는 학습 및 기억을 측정하는 기술을 제시한다. 여기에 제시된 추적 공포 조건화 프로토콜에서 중립 자극과 무조건 자극 사이의 다섯 쌍이있다. 각 조절 재판을 분리하는 20 초 추적 기간이 있습니다. 다음날 동결에 조건 자극 (CS) 및 추적 기간의 프레젠테이션 중에 측정한다. 세 번째 날에 상황에 맞는 메모리를 측정하는 8 분의 시험이있다. 대표적인 결과는 무조건 자극하지 않고 톤 프리젠 테이션을받은 쥐에 비해 혐오 무조건 자극 (충격)로 제시했다 생쥐입니다. 추적 공포 조건화가 성공적으로 다른 공포 조건화 방법이 발견되지 않은 생쥐에 미묘한 학습과 기억 적자 및 개선 사항을 감지하는 데 사용되었습니다. 공포 조건화의 유형은 내측 전두엽 피질과 해마 사이의 연결에 의존 할 것으로 생각됩니다. 하나는 현재 논란이 방법은 편도 독립적 인 것으로 생각되어 있는지 여부입니다. 따라서 다른 공포 조건화 시험은 이러한 지연 공포 조건화를 통해 같은 편도체에 의존 학습과 기억 효과를 검토 할 필요가있다.

대표 결과 우리는 중립 자극을받은하지만 무조건 자극 (아무 충격 조건을)받지 않은 쥐에 비해 무조건 자극 (충격 상태)과 중립 자극 짝을받은 컨트롤 C57BL/6J 성인 마우스에서 데이터를 제시한다. 먼저 프로토콜이 올바르게 수행되었는지 여부를 결정하기 위해 이러한 행동 시험을 설정할 때이 조건을 실행하는 것이 중요하다. 도 1의 데이터는 추적 공포 조건화 ?...

Watch this Scientific Journal Video about Trace Fear Conditioning in Mice at JoVE.com

Trace Fear Conditioning in Mice

다음 실험에서 우리는 쥐의 추적 공포 조건화위한 프로토콜을 설명합니다. 연상 메모리의이 유형은 중립 자극과 무조건 자극을 분리 트레이스 기간을 포함한다.

생쥐의 공포 컨디셔닝을 추적

In this experiment we present a technique to measure learning and memory. In the trace fear conditioning protocol presented here there are five pairings ...

Research

JoVE Journal

Behavior

32.6K 조회수.  Baylor University.  다음 실험에서 우리는 쥐의 추적 공포 조건화위한 프로토콜을 설명합니다. 연상 메모리의이 유형은 중립 자극과 무조건 자극을 분리 트레이스 기간을 포함한다. 

비디오: Trace Fear Conditioning in Mice

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 ...

연구

행동분석학

Disrupting Reconsolidation of Fear Memory in Humans by a Noradrenergic &#946;-Blocker

The basic design used in our human fear-conditioning studies on disrupting reconsolidation includes testing over different phases across three consecutive days. On day 1 - the fear acquisition phase, healthy participants are exposed to a series of picture presentations. One picture stimulus (CS1+) is repeatedly paired with an aversive electric stimulus (US), resulting in the acquisition of a fear association, whereas another picture stimulus (CS2-) is never followed by an US. On day 2 - the memory reactivation phase, the participants are re-exposed to the conditioned stimulus without the US (CS1-), which typically triggers a conditioned fear response. After the memory reactivation we administer an oral dose of 40 mg of propranolol HCl, a &#946;-adrenergic receptor antagonist that indirectly targets the protein synthesis required for reconsolidation by inhibiting the noradrenaline-stimulated CREB phosphorylation. On day 3 - the test phase, the participants are again exposed to the unreinforced conditioned stimuli (CS1- and CS2-) in order to measure the fear-reducing effect of the manipulation. This retention test is followed by an extinction procedure and the presentation of situational triggers to test for the return of fear. Potentiation of the eye blink startle reflex is measured as an index for conditioned fear responding. Declarative knowledge of the fear association is measured through online US expectancy ratings during each CS presentation. In contrast to extinction learning, disrupting reconsolidation targets the original fear memory thereby preventing the return of fear. Although the clinical applications are still in their infancy, disrupting reconsolidation of fear memory seems to be a promising new technique with the prospect to persistently dampen the expression of fear memory in patients suffering from anxiety disorders and other psychiatric disorders.

Disrupting Reconsolidation of Fear Memory in Humans by a Noradrenergic &amp;#946;-Blocker

Disrupting reconsolidation is a promising approach to dampen the behavioral expression of fear memory in patients with anxiety disorders or posttraumatic stress disorder. In a series of human fear conditioning studies we showed that disrupting reconsolidation by the noradrenergic &#946;-blocker propranolol is very effective in erasing conditioned fear responding.

노르 아드레날린 β 차단제에 의해 인간의 두려움 메모리의 재 통합을 방해

The basic design used in our human fear-conditioning studies on disrupting reconsolidation includes testing over different phases across three consecutive ...

Protocol for Studying Extinction of Conditioned Fear in Naturally Cycling Female Rats

Extinction of conditioned fear has been extensively studied in male rodents. Recently, there have been an increasing number of studies indicating that neural mechanisms for certain behavioral tasks and response behaviors are different in females and males. Using females in research studies can represent a challenge because of the variation of gonadal hormones during their estrous cycle. This protocol describes well-established procedures that are useful in investigating the role of estrogen in fear extinction memory consolidation in female rats. Phase of the estrous cycle and exogenous estrogen administration prior to extinction training can influence extinction recall 24 hr later. The vaginal swabbing technique for estrous phase identification described here aids the examination and manipulation of naturally cycling gonadal hormones. The use of this basic rodent model may further delineate the mechanisms by which estrogen can modulate fear extinction memory in females.

Gonadal hormones such as estrogen modulate memory formation in a number of experimental paradigms including fear extinction memory. This protocol describes a set of methods for investigating the influence of gonadal hormones specifically during extinction in naturally cycling females, including estrous cycle monitoring and exogenous estrogen administration.

당연히 여성 쥐 사이클링에 에어컨 공포의 멸종 유학을위한 프로토콜

Extinction of conditioned fear has been extensively studied in male rodents. Recently, there have been an increasing number of studies indicating that ...

Quantifying Social Motivation in Mice Using Operant Conditioning

In this protocol, social motivation is measured in mice through a pair of operant conditioning paradigms. To conduct the experiments, two-chambered shuttle boxes were equipped with two operant levers (left and right) and a food receptacle in one chamber, which was then divided from the second chamber by an automated guillotine door covered by a wire grid. Different stimulus mice, rotated across testing days, served as a social stimulus behind the wire grid, and were only visible following the opening of the guillotine door. Test mice were trained to lever press in order to open the door and gain access to the stimulus partner for 15 sec. The number of lever presses required to obtain the social reward progressively increased on a fixed schedule of 3. Testing sessions ended after test mice stopped lever pressing for 5 consecutive minutes. The last reinforced ratio or breakpoint can be used as a quantitative measure of social motivation. For the second paradigm, test mice were trained to discriminate between left and right lever presses in order to obtain either a food reward or the social reward. Mice were rewarded for every 3 presses of each respective lever. The number of food and social rewards can be compared as a measurement of the value placed upon each reward. The ratio of each reward type can also be compared between mouse strains and the change in this ratio can be monitored within testing sessions to measure satiation with a given reward type. Both of these operant conditioning paradigms are highly useful for the quantification of social motivation in mouse models of autism and other disorders of social behavior.

This article describes a new protocol for the quantitative measurement of social motivation in mice using operant conditioning for a social reward. The protocol is useful for the measurement of social motivation in mouse models of autism and other disorders of social behavior.

조건화를 사용하여 마우스 사회적 동기 부여를 정량화

In this protocol, social motivation is measured in mice through a pair of operant conditioning paradigms. To conduct the experiments, two-chambered ...

Whisker-signaled Eyeblink Classical Conditioning in Head-fixed Mice

Eyeblink conditioning is a common paradigm for investigating the neural mechanisms underlying learning and memory. To better utilize the extensive repertoire of scientific techniques available to study learning and memory at the cellular level, it is ideal to have a stable cranial platform. Because mice do not readily tolerate restraint, they are usually trained while moving about freely in a chamber. Conditioned stimulus (CS) and unconditioned stimulus (US) information are delivered and eyeblink responses recorded via a tether connected to the mouse's head. In the head-fixed apparatus presented here, mice are allowed to run as they desire while their heads are secured to facilitate experimentation. Reliable conditioning of the eyeblink response is obtained with this training apparatus, which allows for the delivery of whisker stimulation as the CS, a periorbital electrical shock as the US, and analysis of electromyographic (EMG) activity from the eyelid to detect blink responses.

The preparation presented here for whisker-signaled eyeblink conditioning in head-fixed mice precisely stimulates specific whiskers while allowing mice to ambulate on a cylindrical treadmill. A whisker stimulation conditioned stimulus (CS) paired with a periorbital shock unconditioned stimulus (US) results in reliable associative learning on this apparatus.

머리 고정 쥐의 수염-신호 Eyeblink 클래식 컨디셔닝

Eyeblink conditioning is a common paradigm for investigating the neural mechanisms underlying learning and memory. To better utilize the extensive ...

Reducing State Anxiety Using Working Memory Maintenance

The purpose of this protocol is to explain how to examine the relationship between working memory processes and anxiety by combining the Sternberg Working Memory (WM) and the threat of shock paradigms. In the Sternberg WM paradigm, subjects are required to maintain a series of letters in the WM for a brief interval and respond by identifying whether the position of a given letter in the series matches a numerical prompt. In the threat of shock paradigm, subjects are exposed to alternating blocks where they are either at risk of receiving unpredictable presentations of a mild electric shock or are safe from the shock. Anxiety is probed throughout the safe and threat blocks using the acoustic startle reflex, which is potentiated under threat (Anxiety-Potentiated Startle (APS)). By conducting the Sternberg WM paradigm during the threat of shock and probing the startle response during either the WM maintenance interval or the intertrial interval, it is possible to determine the effect of WM maintenance on APS.

This protocol demonstrates how to measure anxiety-potentiated startle during the Sternberg Working Memory paradigm.

작업 기억 관리를 사용하여 상태 불안감 감소시키기

The purpose of this protocol is to explain how to examine the relationship between working memory processes and anxiety by combining the Sternberg Working ...

Classical Short-Delay Eyeblink Conditioning in One-Year-Old Children

Classical eyeblink conditioning (EBC) refers to the learned association between a conditioned stimulus (an auditory tone) and an unconditioned stimulus (a puff of air to the cornea). Eyeblink conditioning is often used experimentally to detect abnormalities in cerebellar-dependent learning and memory that underlies this type of associative learning. While experiments in adults and older children are relatively simple to administer using commercial equipment, eyeblink conditioning in infants is more challenging due to their poor compliance, which makes correct positioning of the equipment difficult. To achieve conditioning in one-year-old infants, a custom-made or an adapted commercial system can be used to deliver the air puff to the infant's cornea. The main challenge lies in successfully detecting and classifying the behavioral responses. We report that automated blink detection methods are unreliable in this population, and that conditioning experiments should be analyzed using frame-by-frame analysis of supplementary video camera recordings. This method can be applied to study developmental changes in eyeblink conditioning and to examine whether this paradigm can detect children with neurological disorders.

This protocol describes an eyeblink conditioning paradigm appropriate for experiments with one-year-old infants. Commercial or custom-made equipment can be used to deliver the stimuli, and data collection and analysis should be performed on the video recordings.

Classical eyeblink conditioning (EBC) refers to the learned association between a conditioned stimulus (an auditory tone) and an unconditioned stimulus (a ...

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.

스트레스 강화 된 두려움 학습, 외상 후 스트레스 장애의 강력한 설치류 모델

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

Visual Classical Conditioning in Wood Ants

Several species of insects have become model systems for studying learning and memory formation. Although many studies focus on freely moving animals, studies implementing classical conditioning paradigms with harnessed insects have been important for investigating the exact cues that individuals learn and the neural mechanisms underlying learning and memory formation. Here we present a protocol for evoking visual associative learning in wood ants through classical conditioning. In this paradigm, ants are harnessed and presented with a visual cue (a blue cardboard), the conditional stimulus (CS), paired with an appetitive sugar reward, the unconditional stimulus (US). Ants perform a Maxilla-Labium Extension Reflex (MaLER), the unconditional response (UR), which can be used as a readout for learning. Training consists of 10 trials, separated by a 5-minute intertrial interval (ITI). Ants are also tested for memory retention 10 minutes or 1 hour after training. This protocol has the potential to allow researchers to analyze, in a precise and controlled manner, the details of visual memory formation and the neural basis of learning and memory formation in wood ants.

We present a protocol for the classical conditioning of harnessed ants that permits researchers to study visual learning and memory formation at a level of analysis not possible with freely moving individuals.

나무 개미에 비주얼 고전 컨디셔닝

Several species of insects have become model systems for studying learning and memory formation. Although many studies focus on freely moving animals, ...

Fear Incubation Using an Extended Fear-Conditioning Protocol for Rats

Emotional memory has been primarily studied with fear-conditioning paradigms. Fear conditioning is a form of learning through which individuals learn the relationships between aversive events and otherwise neutral stimuli. The most-widely utilized procedures for studying emotional memories entail fear conditioning in rats. In these tasks, the unconditioned stimulus (US) is a footshock presented once or several times across single or several sessions, and the conditioned response (CR) is freezing. In a version of these procedures, called cued fear conditioning, a tone (conditioned stimulus, CS) is paired with footshocks (US) during the training phase. During the first test, animals are exposed to the same context in which training took place, and freezing responses are tested in the absence of footshocks and tones (i.e., a context test). During the second test, freezing is measured when the context is changed (e.g., by manipulating the smell and walls of the experimental chamber) and the tone is presented in the absence of footshocks (i.e., a cue test). Most cued fear conditioning procedures entail few tone-shock pairings (e.g., 1-3 trials in a single session). There is a growing interest in less common versions involving an extensive number of pairings (i.e., overtraining) related to the long-lasting effect called fear incubation (i.e., fear responses increase over time without further exposure to aversive events or conditioned stimuli). Extended fear-conditioning tasks have been key to the understanding of fear incubation&#8217;s behavioral and neurobiological aspects, including its relationship with other psychological phenomena (e.g., post-traumatic stress disorder). Here, we describe an extended fear-conditioning protocol that produces overtraining and fear incubation in rats. This protocol entails a single training session with 25 tone-shock pairings (i.e., overtraining) and a comparison of conditioned freezing responses during context and cue tests 48 h (short-term) and 6 weeks (long-term) after training.

We describe an extended fear-conditioning protocol that produces overtraining and fear incubation in rats. This protocol entails a single training session with 25 tone-shock pairings (i.e., overtraining) and a comparison of conditioned freezing responses during context and cue tests 48 h (short-term) and 6 weeks (long-term) after training.

쥐에 대 한 확장 된 공포 컨디셔닝 프로토콜을 사용 하 여 공포 잠복

Emotional memory has been primarily studied with fear-conditioning paradigms. Fear conditioning is a form of learning through which individuals learn the ...