Name: testing animal anxiety in rats: effects of open arm ledges and closed arm wall transparency in elevated plus maze test
Uploaded: 2018-06-29T12:30:00.000+00:00
Duration: 7 min 54 s
Description: Watch this Scientific Journal Video about Testing Animal Anxiety in Rats: Effects of Open Arm Ledges and Closed Arm Wall Transparency in Elevated Plus Maze Test at JoVE.com

저자는 사용자 지정된 장치를 생산에 그의 원조에 대 한 씨 昭 히라노를 감사 하 고 싶습니다. 저자는 그들의 기술 지원에 대 한 다카하시 고스케 씨와 씨 신고 나카지마를 감사 하 고 싶습니다. 이 작품은 화학 물질의 위험, 건강 및 실험실 과학 연구 보조금, 보건, 노동 및 복지, 일본 (H25-과학-IPPANN-003) 및 연구소의 과학 및 기술 메이지 연구 프로젝트 교부 금 A에 대 한 연구에 의해 지원 되었다 대학, 일본입니다.

여기에 설명 된 모든 실험 절차 승인 기관 동물 관리 및 사용 위원회 메이지 대학의 (IACUC 14-0002(1)). 동물 대표 결과에 사용 하는 다음과 같이 준비 되었다: 남성 Sprague-Dawley 쥐 실험 동물 회사에서 구입 하 고 시작 하기 전에 적어도 1 주일에 대 한 표준 쥐 감 금 소 (W200 × D410 × H250 m m) 당 4 동물의 그룹에서 지 내게 했다는 행동 실험입니다. 동물 주택 방 (12: 12와 명암 일정 조명에 10:00 h), 실내 온도 (25.0 ± 0.5 ° C), 습도 (55 ± 10%), 조명과 환기에 대 한 표준 대기 조건에서 유지 되었다 (10 회 / h), 그리고 동물 식품 (ad libitum 접근 했다 오리엔탈 효 모 주식 회사, 도쿄, 일본) 및 물. 동물 종이 침구 (알파-dri, LSG Corporatin, 도쿄, 일본)와 함께 표준 쥐 (W410 x D250 x H200 m m, Toyoriko 주식 회사, 아이치 현, 일본)에 유지 했다.
1. 높은 플러스 미로 테스트
<ol><li>설정 높은 플러스 미로 (W1, 500 × 500 × H2, 000 mm D1) 사용자 지정된 컨테이너에 장치 (그림 1A) 더하기 모양의 금속 프레임 워크 실행을 지원 하기 위해 바닥 위에 500 m m 방법 무기 (그림 1B)에 컨테이너 천정에 조명 밝게 필요한 빛의 강도 (그림 1C), 및 동작 테스트 (그림 1C)의 라이브 비디오 모니터링을 사용 하도록 중앙 천장에 디지털 비디오 카메라에는 미로의 표면.</li>
	<li>더하기 모양의 프레임 워크에 무기 (L500 x W100 m m)를 수정 하 고 다음 4 개의 팔의 중앙에 광장 중앙 플랫폼 (W100 x D100 m m) 수정 (그림 1D -F). 참고: 우리의 실험실에서 우리는 두 팔을 벌려 (L500 x W100 m m)의 두 종류와 닫힌된 팔의 두 가지 유형 (W100 x H450 mm x L500) (그림 1D). 닫힌된 팔 투명 하거나 불투명 한 벽 (그림 1D), 그리고 열려있는 팔 짧은 난 간 (H5 m m, 그림 1E) 없이 설계 되었습니다. 광장 중앙 플랫폼 (W100 × D100 mm)는 4 개의 팔의 가운데에 배치 됩니다 (그림 1D -F). 따라서, 팔의 다음 네 가지 조합 실험 목적에 따라 준비 될 수 있다: 오픈 선반과 무기 + 투명 한 벽 (선반/투명), 팔 레와 + 불투명 벽 (선반/불투명), 닫힌된 팔 닫힌된 팔 열 팔 ledges + 투명 한 벽 (No-선반/투명), 닫힌된 무기와 선반 없이 두 팔을 벌려 + 닫힌된 팔 불투명 하지 않고 (No-선반/불투명) 벽. 팔 활주로의 표면 행동 분석 제 2에서 설명한 대로 자동된 동작 분석 소프트웨어에 의해 추적 하는 더 나은 동물 있도록 검은색 이다.</li>
	<li>각 팔과 중앙 플랫폼의 원심 끝 부분의 약 100 럭 스 표면에 조명 강도 조정 합니다.</li>
	<li>각 테스트 전에 70% 에탄올 솔루션 전체 기구를 청소 하 고 증류수로 닦아.</li>
	<li>이미징 분석 섹션 2, 행동 분석에서에서 설명 하는 비디오에 대 한 쥐의 행동을 기록 하는 디지털 비디오 카메라를 시작 하는 하드 디스크 레코더에 "녹화 버튼"을 클릭 합니다.</li>
	<li>오픈 팔을 직면 하 고 중앙 플랫폼에서 동물을 놓고 5 분에 대 한 기구에 무료 탐사를 허용 합니다.</li>
	<li>녹음 완료 하드 디스크 레코더에서 "stop 버튼"을 클릭 합니다.</li>
	<li>시험된 동물 동물 주택 룸에 돌아온 배설물 및 urinations의 수를 계산 합니다.</li>
	<li>1.4 단계에서 실험의 끝까지 반복 합니다. 참고: 건조 테스트 실행은 항상 실행 체크는 모든 실험 설정을 올바르게 설정 되어 전에 어떤 실험 동물 실험 조건에 노출 되는 각 실험을 시작 하기 전에. 그렇지 않으면, 일부 동물, 특히 첫 번째 동물 다른 환경 조건 (예를 들어, 아마도 조작 및 동물 단서 가능성이 이전 시험된 동물에 의해 왼쪽 동안는 실험에 의해 유도 된 소음) 발생할 수 있습니다. 참고:는 경험 컨테이너 근처 동물 테스트 되 고 있습니다. 따라서, 비록는 경험 보이지 않습니다 쥐 여전히 수 있습니다 냄새 / 소리를 통해 그들을.</li>
</ol>2. 행동 분석
참고: 행동 분석 수동 관측에 의해 수행 되 고 자동 분석 소프트웨어.
<ol><li>
수동 관측
	<ol>
<li>동물 선반에 배치 후 다음 관찰: 번호 항목 (팔에 항목 정의 모든 인스턴스로 모든 4 발 팔에 있을 때)의 각 팔에 소요 되는 시간 및 오픈 팔을 직면 수 뻗어 참석의 자세 닫힌된 팔을 향해 중앙 플랫폼에서 백업 하는 동안.</li>
		<li>기간 및 스톱 워치 및 컨테이너의 문을 폐쇄 때부터 시작 하는 5 분에 대 한 수에 의해 위에 정의 된 행동의 주파수를 측정 합니다.</li>
	</ol>
</li>
	<li>
자동된 분석
	<ol>
<li>사용 가능한 소프트웨어 프로그램을 사용 하 여 다음 기록: 팔 (중앙 플랫폼에서 근 위, 중간, 그리고 원심 세그먼트)에 여행, 시간에 보낸된 다른 위치 거리 및 시간 별로 분 분석 오픈에 폐쇄 무기입니다.</li>
		<li>수동 관측 시작으로 동시에 자동된 분석 소프트웨어의 "시작 버튼"을 클릭 (녹음 중지 됩니다 자동으로 설정 된 기간을 기록 하는 경우). 참고:이 연구에서 사용 되는 행동 분석 소프트웨어 배경 필드;와 대조를 통해 동물을 추적 고정된 크기 할당, 동물, 밝은 개체로 식별 하 고 그것의 중심점의 운동 따라서 동물 추적에 사용할 수 있습니다.</li>
	</ol>
</li>
</ol>

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저자는 공개 없다.

정신 장애, 불안 장애를 포함 하 여에서 고통 받는 환자의 수는 증가, 그것은 불안의 기본 메커니즘의 이해를 개선 하기 위해 첨단 연구 개발 하는 것이 중요. 이를 위해, 그것은 또한 실험적인 절차를 최적화 하는 것이 중요입니다. 따라서, 하는 상승의 응용 프로그램 플러스 미로 테스트 개선, 우리는 쥐를 사용 하 여 실험적인 결과에 팔 기능의 효과 증명 하고있다.
높은 플러...

기분 장애, 불안 장애, 우울증, 등에서 고통 받는 환자의 수는 증가 하 고 있다. 이러한 장애의 개발 관련 유전과 환경 요인, 비록 자세한 병원 성 기계 장치는 아직 완전히 이해1. 따라서, 연구는 적은 부작용을가지고 효과적인 약물의 개발으로 서 기분 장애의 근본적인 메커니즘을 잠복 근무에 초점을 해야 합니다.
실험실 동물에 대 한 행동 테스트는 뇌 더 높은 기능, 감정 등을 조사 하는 데 유용 합니다. 동물의 행동은 매우 다양, 실험 결과 자주 다릅니다 실험실, 경우에 동일한 타고 난된 긴장 사용된2. 따라서, 연구원은 또한 그들은 실험실에서 안정적이 고 정확한 결과 제공 하기 위해 채택 하는 실험 장치의 기본 기능을 이해 해야 합니다.
높은 플러스 미로 테스트 설치류3,4정서적 반응성 검사 원래 설립 되었다. 전체 기구는 바닥 위에 보통 있는 500 m m 이며 두 오픈 및 두 닫힌된 팔 더하기 모양 수 있도록 이루어져 있다. 팔 탐험 (를 포함 하 여 기간 및 각 팔에 항목의 주파수)의 측정 거리 여행, 및 지정 된 동작 같은 복종된 동물의 뻗어 참석 자세 관측 기간 동안 기록 됩니다. 이러한 조치에 의해 동물 불안 실험 결과로 추정 된다.
높은 플러스 미로 테스트는 세계 전체에서 사용 됩니다 하지만 모든 실험실 사용 같은 장치 디자인5,6, 그리고 이러한 차이 실험 결과 영향을 미칠 수 있습니다. 예를 들어 차이 존재 또는 부재를 들고 따라 ledges의 또는 닫힌된 팔5,6에 대 한 투명 또는 불투명 벽의 사용을 포함 됩니다. 따라 ledges 팔 열고 닫힌된 무기에 대 한 투명 한 벽의 사용은 동물 기구7떨어지를 방지 하 고 닫힌된 팔 각각8,9내부 행동 관찰을 용이 하 게 개발 되었다. 개별 팔 구조 차이 검사 하는 일부 연구 동물 불안1,2,5,10,11 감지 하는 능력에 영향을 미칠 수 있다 지만 , 동물 행동에 (/ 짧은 난 간 없이) 열기 및 폐쇄 (투명/불투명 벽) 팔 기능의 결합된 효과 아직 조사 되지 않는다.
최근 보고서에서 우리는 쥐12불안 같은 행동에 팔 디자인의 이러한 조합의 효과를 논의 했다. 간단히, 오픈 팔 레 크게 증가 쥐, 불안 같은 동작 동안 투명 팔이이 동작 감소도 있습니다. 또한, 높은 검출 감도 없음 ledged 오픈 팔 + ledged 오픈 팔 + 투명 한 벽으로 폐쇄 팔의 조합에 비해 불투명 한 벽으로 폐쇄 팔의 조합에는 완화 약물의 효과의 확인 합니다. 이 보고서에서 우리는 상승에 대 한 우리의 프로토콜을 소개 합니다 플러스 이전 보고서에서 주요 결과의 논의와 함께 테스트, 미로. 우리는이 보고서는 그 누가 고용 또는 누구는 고가 사용 하 여 그들의 연구에서 미로를 계획에 대 한 유용한 정보를 제공할 것입니다 믿습니다.

<table><tbody><tr><td>Customized container (W1500 &amp;times; D1500 &amp;times; H2000 mm)</td><td>NA</td><td>NA</td><td>Handmade by Mr. Akihiro Hirano</td></tr><tr><td>Lighting</td><td>NA</td><td>NA</td><td>Handmade by Mr. Akihiro Hirano</td></tr><tr><td>Elevated plus maze apparatus</td><td>NA</td><td>NA</td><td>Handmade by Mr. Akihiro Hirano</td></tr><tr><td>Digital video camera</td><td>Watec</td><td>WAT-250D2</td><td>Any suitable video camera can be attached to a hard disk recorder for recording tracking profiles.</td></tr><tr><td>Hard disk recorder</td><td>TOSHIBA</td><td>RD-R200</td><td>Any suitable hard disk recorder can be attached to the digital video camera.</td></tr><tr><td>Sprague-Dawley rats</td><td>Charles River Laboratories Japan, Inc.</td><td>NA</td><td>Any suitable rats can be used for your study</td></tr><tr><td>Standard rat cage</td><td>Toyoriko</td><td>TP-105</td><td>Any suitable rat cage be used for your study</td></tr><tr><td>Instrument to measure light intensity</td><td>AS ONE</td><td>1-5818-02</td><td>Any suitable instrument to measure light intensity</td></tr><tr><td>EthoVision XT version 8.0</td><td>Noldus</td><td>NA</td><td>Any suitable software to measure animal behavior</td></tr><tr><td>Diazepam</td><td>Taiyo Pharmaceutical Industry</td><td>1124402A2061</td><td>Any suitable drug can be used for your study</td></tr><tr><td>Tween 80</td><td>Wako Pure Chemical Industries</td><td>9005-65-6</td><td>Any suitable drug can be used for your study</td></tr></tbody></table>

testing animal anxiety in rats: effects of open arm ledges and closed arm wall transparency in elevated plus maze test

높은 플러스 미로 테스트 설치류 동물 불안을 평가 하기 위한 행동입니다. 이 테스트는 행동 과학의 분야에서 널리 적용 됩니다, 하지만 충돌 결과 종종 다른 실험실에서 제공 됩니다. 다른 결과 대 한 이유를 확인 하려면 우리는 이전 실험실, 대부분 특히 존재/부재의 오픈 팔의 측면과 닫힌된 팔 벽의 투명/불투명 함에 ledges 다 팔 기능에 집중 했다. 이전 보고서에서 우리는 그렇지 않으면 동일 실험 조건에서 열리고 닫힌 팔 디자인의 다른 조합에 쥐 행동을 비교 하는 사용자 지정 설계 된 컨테이너를 사용 하 고 팔 기능에 차이 실험을 방해 했다 결과입니다. 간단히, 오픈 팔 레 크게 증가 쥐, 불안 같은 동작 동안 투명 팔이이 동작 감소도 있습니다. 또한, no ledged 오픈 팔 + ledged 오픈 팔 + 투명 한 벽으로 폐쇄 팔의 조합에 비해 불투명 한 벽으로 폐쇄 팔의 조합에는 완화 약물의 효과의 높은 검출 감도 확인 합니다. 이 보고서에서 우리는 상승에 대 한 프로토콜 플러스 미로 테스트, 이전 보고서와 우리의 실험 경험에서 주요 결과의 논의 함께 소개합니다. 우리는이 보고서는 연구자를 고용 또는 누구는 고가 사용 하 여 그들의 연구에서 미로를 계획에 대 한 유용한 정보를 제공할 것입니다 믿습니다.

이전 연구에서 우리는 오픈 팔 레의 결합된 효과 및 미로12에 예비 동작에 닫힌된 팔 벽의 투명도 비교. 쥐가 크게 ledges과 팔에 보다 선반 없이 팔에 훨씬 적은 시간을 보냈다 ([1, 60] F = 13.49, P = 0.0005) (그림 2A). 비슷한 결과 팔에 항목 수에 대 한 또한 찾아냈다 (F [1, 60] 11.17, P = 0.0014 <)12, ledges 존재 팔에 ...

Watch this Scientific Journal Video about Testing Animal Anxiety in Rats: Effects of Open Arm Ledges and Closed Arm Wall Transparency in Elevated Plus Maze Test at JoVE.com

Testing Animal Anxiety in Rats: Effects of Open Arm Ledges and Closed Arm Wall Transparency in Elevated Plus Maze Test

쥐에서 동물 불안을 테스트: 오픈 팔 레와 닫힌된 팔 벽 투명성 높은 플러스 미로 테스트

쥐 불안 같은 동작 테스트는 높은 플러스 미로 테스트 환경에 따라 매우 다릅니다. 여기 우리가 장치 설정, 특히 팔 구조, 미로에 행동에 미치는 영향을 보여줍니다 그리고 우리 완화 약물의 효과 감지 하는 신뢰할 수 있는 프로토콜을 설명. 이 보고서는 불안 연구에 기여할 것입니다.

The elevated plus maze test is a behavioral test for assessing animal anxiety in rodents. Although this test is widely applied in the field of behavioral ...

testing-animal-anxiety-rats-effects-open-arm-ledges-closed-arm-wall

Universidad de Cartagena UDEC

Research

JoVE Journal

Behavior

13.5K 조회수.  Meiji University. 쥐 불안 같은 동작 테스트는 높은 플러스 미로 테스트 환경에 따라 매우 다릅니다. 여기 우리가 장치 설정, 특히 팔 구조, 미로에 행동에 미치는 영향을 보여줍니다 그리고 우리 완화 약물의 효과 감지 하는 신뢰할 수 있는 프로토콜을 설명. 이 보고서는 불안 연구에 기여할 것입니다.

비디오: Testing Animal Anxiety in Rats: Effects of Open Arm Ledges and Closed Arm Wall Transparency in Elevated Plus Maze 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

Barnes Maze Testing Strategies with Small and Large Rodent Models

Spatial learning and memory of laboratory rodents is often assessed via navigational ability in mazes, most popular of which are the water and dry-land (Barnes) mazes. Improved performance over sessions or trials is thought to reflect learning and memory of the escape cage/platform location. Considered less stressful than water mazes, the Barnes maze is a relatively simple design of a circular platform top with several holes equally spaced around the perimeter edge. All but one of the holes are false-bottomed or blind-ending, while one leads to an escape cage. Mildly aversive stimuli (e.g.&#160;bright overhead lights) provide motivation to locate the escape cage. Latency to locate the escape cage can be measured during the session; however, additional endpoints typically require video recording. From those video recordings, use of automated tracking software can generate a variety of endpoints that are similar to those produced in water mazes (e.g.&#160;distance traveled, velocity/speed, time spent in the correct quadrant, time spent moving/resting, and confirmation of latency). Type of search strategy (i.e.&#160;random, serial, or direct) can be categorized as well. Barnes maze construction and testing methodologies can differ for small rodents, such as mice, and large rodents, such as rats. For example, while extra-maze cues are effective for rats, smaller wild rodents may require intra-maze cues with a visual barrier around the maze. Appropriate stimuli must be identified which motivate the rodent to locate the escape cage. Both Barnes and water mazes can be time consuming as 4-7 test trials are typically required to detect improved learning and memory performance (e.g.&#160;shorter latencies or path lengths to locate the escape platform or cage) and/or differences between experimental groups. Even so, the Barnes maze is a widely employed behavioral assessment measuring spatial navigational abilities and their potential disruption by genetic, neurobehavioral manipulations, or drug/ toxicant exposure.

The dry-land Barnes maze is widely used to measure spatial navigation ability in response to mildly aversive stimuli. Over consecutive days, performance (e.g.&#160;latency to locate escape cage) of control subjects improves, indicative of normal learning and memory. Differences between rats and mice necessitate apparatus and methodology changes that are detailed here.

크고 작은 설치류 모델 바네스 미로 테스트 전략

Spatial learning and memory of laboratory rodents is often assessed via navigational ability in mazes, most popular of which are the water and dry-land ...

Use of the Open Field Maze to Measure Locomotor and Anxiety-like Behavior in Mice

Animal models have proven to be invaluable to researchers trying to answer questions regarding the mechanisms of behavior. The Open Field Maze is one of the most commonly used platforms to measure behaviors in animal models. It is a fast and relatively easy test that provides a variety of behavioral information ranging from general ambulatory ability to data regarding the emotionality of the subject animal. As it relates to rodent models, the procedure allows the study of different strains of mice or rats both laboratory bred and wild-captured. The technique also readily lends itself to the investigation of different pharmacological compounds for anxiolytic or anxiogenic effects. Here, a protocol for use of the open field maze to describe mouse behaviors is detailed and a simple analysis of general locomotor ability and anxiety-related emotional behaviors between two strains of C57BL/6 mice is performed. Briefly, using the described protocol we show Wild Type mice exhibited significantly less anxiety related behaviors than did age-matched Knock Out mice while both strains exhibited similar ambulatory ability.

A protocol is provided to use an Open Field Maze to access general locomotor activity, anxiety and emotionality in a laboratory mouse model.

열기 필드 미로의 사용은 마우스의 운동력과 불안 같은 행동을 측정하기 위해

Animal models have proven to be invaluable to researchers trying to answer questions regarding the mechanisms of behavior. The Open Field Maze is one of ...

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

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

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

더블 H 미로 : 설치류의 강력한 행동 학습에 대한 테스트 및 메모리

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

Quantifying Cognitive Decrements Caused by Cranial Radiotherapy

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

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

두개골 방사선 치료에 의한인지 감소를 Quantifying

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

의학

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

Background: Evaluating mild to moderate cognitive impairment in a global cerebral ischemia (i.e. cardiac arrest) model can be difficult due to poor locomotion after surgery. For example, rats who undergo surgical procedures and are subjected to the Morris water maze may not be able to swim, thus voiding the experiment.
New Method: We established a modified behavioral spontaneous alternation T-maze test. The major advantage of the modified T-maze protocol is its relatively simple design that is powerful enough to assess functional learning/memory after ischemia. Additionally, the data analysis is simple and straightforward. We used the T-maze to determine the rats&#39; learning/memory deficits both in the presence or absence of mild to moderate (6 min) asphyxial cardiac arrest (ACA). Rats have a natural tendency for exploration and will explore the alternate arms in the T-maze, whereas hippocampal-lesioned rats tend to adopt a side-preference resulting in decreased spontaneous alternation ratios, revealing the hippocampal-related functional learning/memory in the presence or absence of ACA.
Results: ACA groups have higher side-preference ratios and lower alternations as compared to control.
Comparison with Existing Method(s): The Morris water and Barnes maze are more prominent for assessing learning/memory function. However, the Morris water maze is more stressful than other mazes. The Barnes maze is widely used to measure reference (long-term) memory, while ACA-induced neurocognitive deficits are more closely related to working (short-term) memory.
Conclusions: We have developed a simple, yet effective strategy to delineate working (short-term) memory via the T-maze in our global cerebral ischemia model (ACA).

This protocol describes the use of a modified T-maze to evaluate functional learning/memory in asphyxia cardiac arrest-induced cerebral ischemia.

심장 마비 후 기능 메모리 결과 평가 하기 위해 수정 된 T-Maze를 활용 하 여

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

Elevated Plus Maze for Mice

Although the mouse genome is now completely sequenced, the functions of most of the genes expressed in the brain are not known. The influence of a given gene on a specific behavior can be determined by behavioral analysis of mutant mice. If a target gene is expressed in the brain, behavioral phenotype of the mutant mice could elucidate the genetic mechanism of normal behaviors. The elevated plus maze test is one of the most widely used tests for measuring anxiety-like behavior. The test is based on the natural aversion of mice for open and elevated areas, as well as on their natural spontaneous exploratory behavior in novel environments. The apparatus consists of open arms and closed arms, crossed in the middle perpendicularly to each other, and a center area. Mice are given access to all of the arms and are allowed to move freely between them. The number of entries into the open arms and the time spent in the open arms are used as indices of open space-induced anxiety in mice. Unfortunately, the procedural differences that exist between laboratories make it difficult to duplicate and compare results among laboratories. Here, we present a detailed movie demonstrating our protocol for the elevated plus maze test. In our laboratory, we have assessed more than 90 strains of mutant mice using the protocol shown in the movie. These data will be disclosed as a part of a public database that we are now constructing.
Visualization of the protocol will promote better understanding of the details of the entire experimental procedure, allowing for standardization of the protocols used in different laboratories and comparisons of the behavioral phenotypes of various strains of mutant mice assessed using this test. 


The elevated plus maze test is one of the most widely used tests for measuring anxiety-like behavior in mice. Here, we present a movie showing the detailed procedures for conducting the test.

마우스에 대한 고가 플러스 메이즈

Although the mouse genome is now completely sequenced, the functions of most of the genes expressed in the brain are not known. The influence of a given ...

생물학

Elevated Plus Maze Test Combined with Video Tracking Software to Investigate the Anxiolytic Effect of Exogenous Ketogenic Supplements

The overall goal of this study is to describe the methodology of the elevated plus maze (EPM) test in combination with a video tracking software. The purpose of the method is to document the effect of various potential anxiolytic treatments on laboratory rodent models. The EPM test is based on the rodents&#39; proclivity toward protected, enclosed dark spaces and unconditioned fear of open spaces and heights, and their innate intense motivation to explore novel environments. The EPM test is a widely used behavioral test for investigating the anxiolytic or anxiogenic responses of rodents given drugs that are known to affect behavior. Observation demonstrating a decreased proportion of time spent on closed arms, an increased proportion of time spent on open arms, a reduced number of entries to closed arms, and an elevated number of entries to open arms measured by the EPM test may reflect reduced anxiety levels. Using this method, the effect of exogenous ketone supplements on anxiety-related behavior is tested in Sprague Dawley (SPD) rats. Exogenous ketone supplements are chronically fed to the rats for 83 days or subchronically and acutely orally gavaged, daily for 7 days, before conducting the EPM test. Behavioral data collection is performed using the SMART video tracking system by a blinded observer at the end of the treatments. The main findings indicate that the EPM test is an effective method to detect the ketone supplement-induced anxiolytic effect and can be considered a sensitive measure to assess changes in anxiety behavior associated with drug- or metabolic-based therapies.

Here, we present a protocol to investigate changes in the anxiety level of rodent animal models. The elevated plus maze (EPM) test, used together with a video tracking software, provides a reliable method to document the effect of various potential anxiolytic treatments in preclinical laboratory scenarios.

The overall goal of this study is to describe the methodology of the elevated plus maze (EPM) test in combination with a video tracking software. The ...

Utilizing a Reconfigurable Maze System to Enhance the Reproducibility of Spatial Navigation Tests in Rodents

Several maze shapes are used to test spatial navigation performance and behavioral phenotypes. Traditionally, each experiment requires a unique maze shape, thus requiring several separate mazes in different configurations. The maze geometry cannot be reconfigured in a single environment to accommodate scalability and reproducibility. The reconfigurable maze is a unique approach to address the limitations, allowing quick and flexible configurations of maze pathways in a repeatable manner. It consists of interlocking pathways and includes feeders, treadmills, movable walls, and shut-off sensors. The current protocol describes how the reconfigurable maze can replicate existing mazes, including the T-shaped, plus-shaped, W-shaped, and figure-eight mazes. Initially, the T-shaped maze was constructed inside a single experimental room, followed by modifications. The rapid and scalable protocol outlined herein demonstrates the flexibility of the reconfigurable maze, achieved through the addition of components and behavioral training phases in a stepwise manner. The reconfigurable maze systematically and precisely assesses the performance of multiple aspects of spatial navigation behavior.

The present protocol describes a reconfigurable maze, a unique system for testing spatial navigation and behavioral phenotypes in rodents. The adaptability of this maze system enables the execution of various experiments in a single physical environment. The ease of structural rearrangement generates reliable and reproducible experimental results.

재구성 가능한 미로 시스템을 활용하여 설치류에서 공간 탐색 테스트의 재현성 향상

Several maze shapes are used to test spatial navigation performance and behavioral phenotypes. Traditionally, each experiment requires a unique maze ...

행동 테스트를 사용하여 Angelman Syndrome의 Murine 모델 검증

Behavioral Characterization of an Angelman Syndrome Mouse Model

This manuscript describes a battery of behavioral tests available to characterize Angelman syndrome (AS)-like phenotypes in an established murine model of AS. We use the rotarod learning paradigm, detailed gait analysis, and nest building test to detect and characterize animal motor impairments. We test animal emotionality in the open field and elevated plus maze tests, as well as the affect in the tail suspension test. When AS mice are tested in the open field test, the results should be interpreted with care, since motor dysfunctions influence mouse behavior in the maze and alter activity scores.
The reproducibility and effectiveness of the presented behavioral tests has already been validated in several independent Uba3a mouse lines with different knockout variants, establishing this set of tests as an excellent validation tool in AS research. Models with the relevant construct and face validity will warrant further investigations to elucidate the pathophysiology of the disease and grant the development of causal treatments.

저자 스포트라이트: Angelman Syndrome 쥐 모델을 검증하기 위한 고도로 재현 가능한 행동 테스트 배터리 

This manuscript presents a set of highly reproducible behavioral tests to validate an Angelman syndrome mouse model.

Angelman 증후군 마우스 모델의 행동 특성

This manuscript describes a battery of behavioral tests available to characterize Angelman syndrome (AS)-like phenotypes in an established murine model of ...