This work was supported by a CIHR operating grant (MOP-102624) to Q. Y. We thank Dr Carolyn Harley for helpful discussions throughout the study, Dr. Qinlong Hou, Amin Shakhawat, and Andrea Darby-King for technical support.

남녀의 스프 라그 돌리 쥐 (찰스 리버) 새끼가 사용된다. 새끼는 (출생 PD0 인) PD1에 12 도태된다. 댐은 음식과 물에 대한 광고 무제한으로 액세스 할 수있는 12 시간 명 / 암주기에 유지됩니다. 실험 절차는 기념 대학의 기관 동물 관리위원회에 의해 승인되었습니다. 1 코 플러그 건설 참고 :.이 절차는 적응 커밍스 외로부터 수정 (1997) 35. <ol><li> Aquire 폴리에틸렌 20 튜브 3-0 실크 봉합사 실. </li><li> 약 0.8 mm에 폴리에틸렌 20 튜브의 작은 조각을 잘라. </li><li> 스레드는 튜브 섹션의 양쪽에 존재하도록 제조 된 튜브를 통해 달기 실크 봉합사. </li><li> 플러그의 외부 실의 한쪽 끝에서 스레드에서 매듭을 묶어. </li><li> 실의 매듭을 통해 아래로 튜브의 섹션을 당깁니다. 매듭은 튜브 내부에 제출해야한다.</li><li> 스레드 ~ 2mm가 튜브의 일단 부로부터 돌출되도록 실의 양단을 트림 (도 1a 참조). </li></ol> 교육 전에 2 나리스 폐쇄 <ol><li> 일반 침구로 덮여 안전한 접시에 댐과 장소에서 강아지를 제거합니다. </li><li> 콧 구멍에 국소 마취제 젤리, 2 % Xylocaine을 가볍게 두 드리하는면 팁 응용 프로그램을 사용하는 폐색합니다. </li><li> 강아지가 ~ 3 분 동안 접시에 휴식을 허용합니다. </li><li> 비 지배적 인 손에 부드럽게 안전하게 강아지를 잡으십시오. </li><li> 지배적 인 손을 사용하여, 코 플러그를 픽업 스레드가 돌출되어 있지 않은부터 팁 주위 같은​​ 국소 마취제 젤리를 두드려. 이 플러그 삽입과 관련된 약간의 통증이 모두 마취제 역할을하고 콧 구멍 안쪽에 윤활제 역할을합니다. 참고 : Xylocaine의 효과는 몇 분 내에 시작하고 20 ~ 30 분 동안 지속됩니다. xylocaine 젤리 사용 후 플러그 삽입의 일반적인 쇼 좋은 내성 새끼 (minimum 어려움을 겪고와 발성). </li><li> 조심스럽게 단단히 강아지를 들고와 플러그가 완전히 삽입 만 2mm 스레드가 콧 구멍에서 돌출 될 때까지 천천히 아주 부드러운 푸시와 플러그를 회전하여 코 플러그를 삽입합니다 (그림 1B 참조). 이 과정에서 두 콧 구멍에서 더 출혈이 없어야합니다. 코 플러그 삽입시 출혈 새끼 제외하고 댐으로 돌아갑니다 </li><li> 동물이 플러그에 길들하기 위해 5 분 동안이 접시에 휴식을 허용합니다. </li><li> 습관화 요리에서 강아지를 제거하고 에어컨 패러다임 (24)를 시작합니다. </li></ol><img alt="그림 1" fo:content-width="5in" src="/files/ftp_upload/51808/51808fig1highres.jpg" width="500" /> 코 플러그의 그림 1 건설. A) 회로도 코 플러그의 제조 단계를 도시. 스레드는 폴리에틸렌 튜브를 당겨; 매듭이 만들어 PUL된다이를 차단하는 튜브 중앙으로 이끄는; 두 끝)가 호스. B 중 한 쪽 끝에서 2mm의 잔류와 함께 전면 및 한 콧 구멍에 코 플러그가있는 쥐의 측면보기를 잘립니다. 3 향기 침구 준비 <ol><li> 새 장갑을 착용하고, 악취 오염을 방지 비닐 봉지에 우드 칩 침구 500 ㎖에 배치 할 흄 후드. </li><li> 0.3 ml의 박하 추출물을 그릴 주사기를 사용하고, 비닐 봉투에있는 침구를 통해이 스프레이. </li><li> , 가방 차단 타이 적극적으로 가방을 흔들어 침구는 5 분 동안 가방에 휴식을 할 수 있습니다. </li><li> 사용하기 전에 5 분 동안 흄 후드에서 발견 명확한, 얕은, 아크릴 훈련 상자 (20 × 20 × 5cm 3, 그림 2A)에서 냄새가 침대를 배치합니다. 침구가​​ 준비되면,이 장갑을 폐기하고,이 장갑은 동물과 접촉하지 않도록하십시오. </li><li> 동일한 투명한 플라스틱 상자에 무향 침대를 놓고 엉이 향수 침구 또는 사용 장갑에 접촉하지 않는지 확인하십시오. </li></ol> 4 냄새 컨디셔닝 패러다임 (그림 2A에서 그림 참조) 새끼는 PD 6, 또는 여러 시험 세션 (하루에 하나의 세션, PD 3-6)에, 하나 하나의 조화 세션을받을. <ol><li> 냄새가 침대에 길들여 강아지를 놓습니다. 제어 냄새 만 (O / S -)의 경우 새끼, 다음 4.5 단계로 이동, 10 분 동안 침대에서이 새끼를 둡니다. 실험 냄새 + 스트로크 (O / S +) 새끼는,이 섹션에서 다음 단계를 계속합니다. </li><li> 스트로크 30 초 동안 강아지는 작은 붓을 사용. 주로 강아지의 뒷다리 지역 주위에 빠른 원형 동작을 사용합니다. </li><li> 강아지가 30 초 동안 휴식을 허용합니다. </li><li> 반복 4.2 및 10 분의 총 4.3 (쓰다듬어 + 냄새 즉, 10 페어링) 단계를 반복합니다. </li><li> 컨디셔닝 상자에서 강아지를 제거 코 플러그를 제거하고 댐에 강아지를 반환합니다. </li></ol> &lt;P 클래스 = &quot;jove_title&quot;&gt; 5. 편측 냄새 환경 테스트 (그림 2B에서 그림 참조) 테스트는 최종 트레이닝 세션 다음 다양한 시점 (예를 들면, 24 또는 48 시간)에서 발생한다. 테스트는 2cm 중성 영역으로 구분이 훈련 박스 (훈련 박스 3.4에서 설명된다)의 상단에 배치되는 스테인리스 테스트 실 (30 X 20 X 18cm 3)에서 수행된다. 다른 상자가 깨끗하고, 무향의 침대를 포함하는 동안 하나의 훈련 상자가 박하 향기 침구가 포함되어 있습니다. 테스트 챔버의 바닥은 플라스틱 메쉬 분리형 시트 (도 2b)에 의해 커버되는 금속 격자이다. <ol><li> 한 박하와 제 3 당 한 무향 침구를 준비하고, 2cm 간격 테스트 챔버의 양측에서 각 상자를 배치합니다. 테스트 챔버의 금속 격자 층에있는 플라스틱 메시를 놓습니다. </li><li> 댐에서 강아지를 제거하고 콧 구멍에 무취 실리콘 그리스의 회사 명인를 배치그는 훈련 기간 동안 폐색된다. 재 적용 필요에 따라 제 1 검사 절차 내내 그리스. 참고 : 교육 및 테스트하는 동안 임의 콧 구멍 폐쇄는 편견을 피하기 위해 고려 될 수있다. </li><li> 테스트 챔버의 중립 영역에 강아지를 놓습니다. </li><li> 강아지가 챔버의 양측 지출 한 시간을 기록, 강아지가 1 분 동안 챔버를 탐험 할 수 있도록 허용 (예 : 박하 또는 중립적 향기 침구 이상). </li><li> 강아지가 덮여 플라스틱 보유 챔버에서 1 분 동안 휴식을 허용합니다. </li><li> 반복은 10 분의 총 5.2 &amp; 5.3 단계 (즉, 5 나머지 실험에 의해 분리 실험 5 실험) 방향 선호도에 대해 제어하기 위해 챔버 내에서 강아지의 초기 방향 전환. </li><li> 즉시 시험 다음, 콧 구멍에서 그리스를 닦아. </li><li> 2 절에 따라 반대 콧 구멍에 폴리에틸렌 noseplug를 삽입하고 동물이 10 분 동안 휴식을 할 수 있습니다. </li><li> 에서와 같이 다시 한번 강아지를 테스트<html> 5.3 -5.6는 플러그를 제거하고 댐에 강아지를 반환합니다. 제거하고 95 % 에탄올로 테스트 챔버의 플라스틱 메쉬를 청소하고 액체가 증발 할 수있다. 다음 강아지를 테스트하기 전에 다시 메시를 놓습니다. 참고 : 테스트 중 첫 번째 콧 구멍 폐쇄에 실리콘 그리스를 적용 출혈과 코의 플러그 삽입과 관련된 스트레스의 가능성을 방지 할 수 있습니다. </li></ol><img alt="그림이" fo:content-width="5in" src="/files/ftp_upload/51808/51808fig2highres.jpg" width="500" /> 그림 2 초기 악취 환경 교육 및 테스트. 패러다임을 쓰다듬어 + 냄새를 사용하여 A) 초기 악취 환경 교육. B) 한면에 박하 침구와 함께 두 개의 선택 냄새 환경 테스트, 반대편에 무향 침대를 제어 할 수 있습니다. 2cm 중립 영역은 사이에 배치됩니다. (6) 단일 나리스 폐쇄의 효과를 테스트 NT는 &quot;&gt;이 실험은 하나의 콧 구멍 폐쇄가 후각 시스템의 편측 활성화로 연결 여부를 결정하기 위해 수행된다. <ol><li> 2 절에 설명 된대로 PD 6 또는 7 새끼에 일방적 콧 구멍의 폐색을 수행합니다. </li><li> ~ 5 분의 습관화 후, 해당 플라스틱 용기에 강아지를 배치하고 10 분 동안 조직의 조각에 담가 30 ㎕의 순수 박하유에 노출. </li><li> 즉시 박하 냄새에 노출 된 후, 복강 내 (IP) 클로 랄 전신 마취로 수화물 (400 ㎎ / ㎏), 또는 pentobarbitol (150 ㎎ / ㎏)와 강아지를 주입. </li><li> 완전히 (꼬리 부분 또는 발 핀치에 대한 응답을 보여주는) 마취 후 transcardially 0.1 M 인산 완충 용액, PBS에 용해 파라 포름 알데히드 (4 % 다음에 ~ 1 분 식염수 (0.9 %)의 얼음처럼 차가운 솔루션 강아지, 관류 ). </li><li> 다음 트랜스, 파라 포름 알데히드 관류 10 분 후 뇌를 수집하고 4 ° C에서 하룻밤 파라 포름 알데히드에 배치추가로 24 시간 동안 자당 용액 (PBS에서 20 %)로 뇌를 FER. </li><li> 저온 유지 장치와 30 μm의 두께로 관상 뇌 조각을 잘라. OB와 PC 조각을 수집하고 pCREB 항체 21,25,30 표준 면역 조직 화학 염색 뒤에 젤라틴 코팅 슬라이드에 탑재합니다. </li></ol> 7 단일 나리스 폐쇄의 가역성 테스트 차단 효과는 코 플러그를 제거 다음 24 시간에서 뒤집을 수 있는지 여부를이 실험 테스트합니다. <ol><li> 2 절에 설명 된대로 PD 6 또는 7 새끼에 일방적 콧 구멍의 폐색을 수행합니다. </li><li> (훈련 중에 콧 구멍 폐쇄의 기간에 해당 - 5 분​​의 습관화 + 10 분 교육) 15 분 후, 코 플러그를 제거하고 댐에 강아지를 반환합니다. </li><li> 6.2에 설명 된대로 24 시간 후, 10 분 동안 덮여 플라스틱 용기에 박하 냄새에 강아지를 노출합니다. </li><li> 종파에서 동일한 단계를 수행6.3-6.6를 이온. </li></ol>

<ol>
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The authors have nothing to disclose.

중요한 시간 창 내 쥐 새끼의 편측 냄새 학습과 기억 모델은 첫번째 홀과 동료들에 의해 설립되었습니다. 연구 33,34,36의 시리즈, 그들은 냄새 기본 메모리가 쥐 새끼에서 PD 6에 하나의 콧 구멍으로 냄새 + 우유 짝에 의해 편측 될 수 있음을 보여 주었다. 기본 설정 메모리는 같은 콧 구멍 교육 및 시험 동안 열려있을 때 강력한했지만, 폐쇄 된 콧 구멍이 차단을 해제하고 테스트 때 관찰되지....

후각은 성공적으로 탐색하거나 자신의 환경에서 살아남을 수 없을 것이다없이 설치류의 기본 감각 양상이다. 그것은 일을 공급하기 위해 자신의 어머니를 찾기 위해 후각을 사용하여, 첫번째 산후 주 동안 참조되지도들을 수 있습니다 둘 신생아 새끼에 특히 중요합니다. 결과적으로, 신생아 래트 새끼가 간단한 조작으로 실험을 선호하는 냄새 조절 될 수있다. 자극의 다양한 중첩 환경 2,3, 우유, 유아 4-6, 쓰다듬어 촉각 자극 7-을 포함하여 신생아에서 새로운 냄새에 대한 조건 반응 (조건 자극, CS)를 유도하는 무조건 자극 (UCS)로 사용되어왔다 12, 꼬리 핀치 (13), 산모의 타액 13, 가벼운 발 충격 14-18, 두개 내 뇌 자극 19. 본 연구는 잘 설립 초기 냄새 환경 패러다임을 고용하고있어서,이 경우 박하의 냄새,, 난박하 24 시간 후 10,11,20에 대한 선호를 생산하기 위해 촉각 자극과 결합 될 수있다. 이 냄새의 기억은 주로 후각 전구 (OB) 21 ~ 23과 전방 배 모양의 피질 (APC) (24, 25)를 포함하여, 그대로 후각 회로에 따라 달라집니다. 초기 냄새 환경 학습의 실험 연구 심화 및 포유 동물 메모리의 분자 및 생리 학적 토대에 대한 우리의 이해를 넓혔다. 이 포유 동물 모델은 메모리 메커니즘을 연구하는 여러 가지 장점이 있습니다. 먼저, UCS 신호의 소스는 신경 확인되었다. 위에서 언급 한 바와 같이 다양한 자극 차례로 22,27,28 학습 지원 세포 및 생리 학적 효과를 일으키는 원인이되는 OB와 APC에서 여러 아드레날린 수용체를 활성화 궤적 coeruleus 노르 에피네프린 릴리스 26를 자극한다. 둘째, 메모리지지 메커니즘은 잘 정의 된 층류 신경 구조에서 일어난다.신생아 쥐의 후각 회로의 단순 시냅스 가소성과 관련된 복잡한 프로세스를 발견 할 수와 이상적인 프레임 워크와 연구자를 제공합니다. 다른 구조 29 중 OB와 ipsilaterally 측면 후각 기관 (LOT)를 통해 배 모양의 피질 (PC) 차례 프로젝트에서 이러한 승모판 / 술 세포에서 승모판 / 술 세포 상에 후각 상피 프로젝트의 후각 감각 뉴런 (OSN). OSN의 OB (30, 31)에서 시냅스와 LOT는 APC에서 (24, 25)을 시냅스 모두 시냅스 학습 지원 변경 및 메모리에 중요한 서식처로 확인되었습니다. 셋째, 쥐의 초기 시대에, 후각 입력은 쉽게 편측 할 수 있습니다. 각 APC는 한 번이 흰색 물질이 완전히 산후 12 일 (PD12) (32)에 형성되고, 전방 교련을 통해 양국 간 냄새 정보에 액세스 할 수 있습니다. PD 12 전에 냄새 입력은 하나의 콧 구멍 폐쇄 24,25,31,33,34 &lt;을 통해 OB와 APC를 ipisilateral하기 위해 분리 할 수있다/ SUP&gt;. 단일 콧 구멍 폐쇄 오픈 콧 구멍에서 냄새가 기억 형성을 허용하고, 12 (33) PD하기 전에 가려 콧 구멍에서 동일한 메모리를 방지 할 수 있습니다. 냄새의 기억은 모두 OB 및 APC를 포함한 동측 반구에 격리됩니다. 따라서 각 쥐 강아지 학습 및 생리학의 토대에 대한 자신의 제어 할 수 있습니다. 본 연구에서는, 편측 이른 악취 기본 학습 프로토콜이 도입된다. 이 방법은 이에 필요한 동물의 수와 일반적인 편차 모두를 줄일 인트라 동물 24,25,31 제어를 제공하여 악취를 뒷받침 신경망 학습 메카니즘을 연구하기위한 강력한 도구로서 기능한다. 나리스 폐쇄는 그리스 나 코 플러그가 적용되는 최소한의 스트레스 또는 동물의 손상으로 제거 할 수 있다는 점에서 가역적이다. 여기에서, 첫째, 초기 악취 환경 교육 및 테스트의 자세한 절차는 번호와 함께 하나의 콧 구멍 폐쇄를 사용하여 편측 프로토콜을 중심으로 설명되어 있습니다전자 플러그. 그런 다음 결과는 냄새 입력을 분리하고 편측 냄새 메모리를 생산하는 하나의 콧 구멍 폐쇄의 효과를 보여주기 위해 제공됩니다. 마지막으로, 모두가 학습 및 지원 메모리 표현을 생성 후각 시스템의 생리적 변화를 연구하기 위해 편측 학습 모델을 사용의 가능성이 논의된다.

<table><tbody><tr><td>Polythylene 20 tubing</td><td>Intramedic</td><td>427406</td><td>Non-radiopaque, Non-toxic</td></tr><tr><td>3-0 Silk suture thread</td><td>Syneture</td><td>Sofsilk</td><td>Non-absorbent</td></tr><tr><td>Silicone grease</td><td>Warner Instrument</td><td>64-0378</td><td>Odorless</td></tr><tr><td>2% Xylocaine gel</td><td>AstraZeneca</td><td>Prod. No 061</td><td>Lidocaine hydrochloride jelly, purchased at local pharmacy</td></tr><tr><td>Paint brush</td><td>Dynasty</td><td>206R</td><td>Similar size/other brands work too</td></tr><tr><td>Peppermint extract</td><td>Sigma-Aldrich</td><td>W284807</td><td>Other brands should be okay too</td></tr><tr><td>Training box</td><td>Custom-made</td><td>N/A</td><td>Acrylic box (20 x 20 x 5 cm&lt;sup&gt;3&lt;/sup&gt;), see &lt;b&gt;Figure 2A&lt;/b&gt;. Parameters and material for the box are not critical and can be modified. Material used should be odorless and does not absorb odors</td></tr><tr><td>Testing chamber</td><td>Custom-made</td><td>N/A</td><td>Stainless steel (30 x 20 x 18 cm&lt;sup&gt;3&lt;/sup&gt;), see &lt;b&gt;Figure 2B&lt;/b&gt;. Parameters and material for the chamber are not critical and can be modified. For example, an acrylic chamber instead of a stainless steel one can be used</td></tr><tr><td>pCREB antibody</td><td>Cell Signaling</td><td>9198</td><td>Ser 133 (87G3) Rabbit mAb</td></tr><tr><td>Chloral hydrate</td><td>Sigma-Aldrich</td><td>C8383</td><td>N/A</td></tr><tr><td>Paraformaldehype</td><td>Sigma-Aldrich</td><td>P6148</td><td>N/A</td></tr><tr><td>Sucrose</td><td>Sigma-Aldrich</td><td>S9378</td><td>N/A</td></tr></tbody></table>

a lateralized odor learning model in neonatal rats for dissecting neural circuitry underpinning memory formation

Rat pups during a critical postnatal period (&#8804; 10 days) readily form a preference for an odor that is associated with stimuli mimicking maternal care. Such a preference memory can last from hours, to days, even life-long, depending on training parameters. Early odor preference learning provides us with a model in which the critical changes for a natural form of learning occur in the olfactory circuitry. An additional feature that makes it a powerful tool for the analysis of memory processes is that early odor preference learning can be lateralized via single naris occlusion within the critical period. This is due to the lack of mature anterior commissural connections of the olfactory hemispheres at this early age. This work outlines behavioral protocols for lateralized odor learning using nose plugs. Acute, reversible naris occlusion minimizes tissue and neuronal damages associated with long-term occlusion and more aggressive methods such as cauterization. The lateralized odor learning model permits within-animal comparison, therefore greatly reducing variance compared to between-animal designs. This method has been used successfully to probe the circuit changes in the olfactory system produced by training. Future directions include exploring molecular underpinnings of odor memory using this lateralized learning model; and correlating physiological change with memory strength and durations.

여기에서 우리는 냄새 입력을 분리하고 하나의 반구 학습에 콧 구멍 폐쇄의 효과 및이 방법의 가역성을 보여주기 위해 이전에 설정된 결과 (24)의 일부를 검토합니다. 초기 냄새 환경 설정 훈련 도중 단일 콧 구멍 폐쇄는 편측 냄새 메모리 (24)에 연결됩니다. 메모리 스페어 나리스 (그림 3)으로 제한됩니다. 강아지가 훈련 도중으로 폐색 같은 콧 구...

Watch this Scientific Journal Video about A Lateralized Odor Learning Model in Neonatal Rats for Dissecting Neural Circuitry Underpinning Memory Formation at JoVE.com

A Lateralized Odor Learning Model in Neonatal Rats for Dissecting Neural Circuitry Underpinning Memory Formation

This protocol introduces lateralized early odor preference learning in rats using acute single naris occlusion. Lateralized learning permits the examination of behavioral outcomes and underpinning biological mechanisms within the same animals, reducing variance induced by between-animal designs. This protocol can be used to investigate molecular mechanisms underpinning early odor learning.

신경 회로 토대 메모리 형성을 해부에 대한 신생아 흰쥐의 편측 냄새 학습 모델

Rat pups during a critical postnatal period (&#8804; 10 days) readily form a preference for an odor that is associated with stimuli mimicking maternal ...

a-lateralized-odor-learning-model-neonatal-rats-for-dissecting-neural

Universidad de Cartagena UDEC

Research

JoVE Journal

Neuroscience

8.9K 조회수.  Faculty of Medicine, Memorial University. This protocol introduces lateralized early odor preference learning in rats using acute single naris occlusion. Lateralized learning permits the examination of behavioral outcomes and underpinning biological mechanisms within the same animals, reducing variance induced by between-animal designs. This protocol can be used to investigate molecular mechanisms underpinning early odor learning.

비디오: A Lateralized Odor Learning Model in Neonatal Rats for Dissecting Neural Circuitry Underpinning Memory Formation

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 ...

연구

행동분석학

A Method for Remotely Silencing Neural Activity in Rodents During Discrete Phases of Learning

This protocol describes how to temporarily and remotely silence neuronal activity in discrete brain regions while animals are engaged in learning and memory tasks. The approach combines pharmacogenetics (Designer-Receptors-Exclusively-Activated-by-Designer-Drugs) with a behavioral paradigm (sensory preconditioning) that is designed to distinguish between different forms of learning. Specifically, viral-mediated delivery is used to express a genetically modified inhibitory G-protein coupled receptor (the Designer Receptor) into a discrete brain region in the rodent. Three weeks later, when designer receptor expression levels are high, a pharmacological agent (the Designer Drug) is administered systemically 30 min prior to a specific behavioral session. The drug has affinity for the designer receptor and thus results in inhibition of neurons that express the designer receptor, but is otherwise biologically inert. The brain region remains silenced for 2-5 hr (depending on the dose and route of administration). Upon completion of the behavioral paradigm, brain tissue is assessed for correct placement and receptor expression. This approach is particularly useful for determining the contribution of individual brain regions to specific components of behavior and can be used across any number of behavioral paradigms.

This protocol describes how to temporarily and remotely silence neuronal activity in discrete brain regions while rats are engaged in learning and memory tasks. The approach combines pharmacogenetics (Designer-Receptors-Exclusively-Activated-by-Designer-Drugs) with a behavioral paradigm (sensory preconditioning) that is designed to distinguish between different components of learning.

학습의 개별 단계에서 설치류에서 원격 음소거 신경 활동하는 방법

This protocol describes how to temporarily and remotely silence neuronal activity in discrete brain regions while animals are engaged in learning and ...

Appetitive Associative Olfactory Learning in Drosophila Larvae

In the following we describe the methodological details of appetitive associative olfactory learning in Drosophila larvae. The setup, in combination with genetic interference, provides a handle to analyze the neuronal and molecular fundamentals of specifically associative learning in a simple larval brain.
Organisms can use past experience to adjust present behavior. Such acquisition of behavioral potential can be defined as learning, and the physical bases of these potentials as memory traces1-4. Neuroscientists try to understand how these processes are organized in terms of molecular and neuronal changes in the brain by using a variety of methods in model organisms ranging from insects to vertebrates5,6. For such endeavors it is helpful to use model systems that are simple and experimentally accessible. The Drosophila larva has turned out to satisfy these demands based on the availability of robust behavioral assays, the existence of a variety of transgenic techniques and the elementary organization of the nervous system comprising only about 10,000 neurons (albeit with some concessions: cognitive limitations, few behavioral options, and richness of experience questionable)7-10.
Drosophila larvae can form associations between odors and appetitive gustatory reinforcement like sugar11-14. In a standard assay, established in the lab of B. Gerber, animals receive a two-odor reciprocal training: A first group of larvae is exposed to an odor A together with a gustatory reinforcer (sugar reward) and is subsequently exposed to an odor B without reinforcement 9. Meanwhile a second group of larvae receives reciprocal training while experiencing odor A without reinforcement and subsequently being exposed to odor B with reinforcement (sugar reward). In the following both groups are tested for their preference between the two odors. Relatively higher preferences for the rewarded odor reflect associative learning - presented as a performance index (PI). The conclusion regarding the associative nature of the performance index is compelling, because apart from the contingency between odors and tastants, other parameters, such as odor and reward exposure, passage of time and handling do not differ between the two groups9.

Appetitive Associative Olfactory Learning in Drosophila Larvae

Drosophila larvae are able to associate odor stimuli with gustatory reward. Here we describe a simple behavioral paradigm that allows the analysis of appetitive associative olfactory learning.

에 Appetitive 연관 강한 학습<em&gt; Drosophila</em&gt; 애벌레

In the following we describe the methodological  details of appetitive associative olfactory learning in Drosophila larvae. The setup, in combination with ...

신경과학

Drosophila Adult Olfactory Shock Learning

Drosophila have been used in classical conditioning experiments for over 40 years, thus greatly facilitating our understanding of memory, including the elucidation of the molecular mechanisms involved in cognitive diseases1-7. Learning and memory can be assayed in larvae to study the effect of neurodevelopmental genes8-10 and in flies to measure the contribution of adult plasticity genes1-7. Furthermore, the short lifespan of Drosophila facilitates the analysis of genes mediating age-related memory impairment5,11-13. The availability of many inducible promoters that subdivide the Drosophila nervous system makes it possible to determine when and where a gene of interest is required for normal memory as well as relay of different aspects of the reinforcement signal3,4,14,16.
Studying memory in adult Drosophila allows for a detailed analysis of the behavior and circuitry involved and a measurement of long-term memory15-17. The length of the adult stage accommodates longer-term genetic, behavioral, dietary and pharmacological manipulations of memory, in addition to determining the effect of aging and neurodegenerative disease on memory3-6,11-13,15-21.
Classical conditioning is induced by the simultaneous presentation of a neutral odor cue (conditioned stimulus, CS+) and a reinforcement stimulus, e.g., an electric shock or sucrose, (unconditioned stimulus, US), that become associated with one another by the animal1,16. A second conditioned stimulus (CS-) is subsequently presented without the US. During the testing phase, Drosophila are simultaneously presented with CS+ and CS- odors. After the Drosophila are provided time to choose between the odors, the distribution of the animals is recorded. This procedure allows associative aversive or appetitive conditioning to be reliably measured without a bias introduced by the innate preference for either of the conditioned stimuli. Various control experiments are also performed to test whether all genotypes respond normally to odor and reinforcement alone.

Drosophila Adult Olfactory Shock Learning

The method to measure adult Drosophila associative memory is described. The assay is based on the ability of the fly to associate an odor presented with a negative reinforcer (electric shock) and then recall this information at a later time, allowing memory to be measured.

초파리 성인 후각 충격 학습

Drosophila have been used in classical conditioning experiments for over 40 years, thus greatly facilitating our understanding of memory, including the ...

A Low-Cost, Odor-Reward Association Task for Tests of Learning and Memory

Robust and simple behavioral paradigms of appetitive, associative memory are crucial for researchers interested in cellular and molecular mechanisms of memory. In this paper, an effective and low-cost mouse behavioral protocol is described for examining the effects of physiological manipulation (such as the infusion of pharmacological agents) on the learning rate and duration of odor-reward memory. Representative results are provided from a study examining the differential role of tyrosine kinase receptor activity in short-term (STM) and long-term memory (LTM). Male mice were conditioned to associate a reward (sugar pellet) with one of the two odors, and their memory for the association was tested 2 or 48 h later. Immediately prior to the training, a tyrosine kinase (Trk) receptor inhibitor or vehicle infusions were delivered into the olfactory bulb (OB). Although there was no effect of the infusion on the learning rate, blockade of the Trk receptors in the OB selectively impaired LTM (48 h), and not short-term memory (STM; 2 h). The LTM impairment was attributed to the diminished odor selectivity as measured by the length of the digging time. The culmination of the results of this experiment showed that Trk receptor activation in the OB is the key in olfactory memory consolidation.

An odor-reward, associative learning task was used to investigate the differential effects of physiological manipulation on long-term and short-term memory.

Robust and simple behavioral paradigms of appetitive, associative memory are crucial for researchers interested in cellular and molecular mechanisms of ...

The Olfactory System as a Model to Study Axonal Growth Patterns and Morphology In Vivo

The olfactory system has the unusual capacity to generate new neurons throughout the lifetime of an organism. Olfactory stem cells in the basal portion of the olfactory epithelium continuously give rise to new sensory neurons that extend their axons into the olfactory bulb, where they face the challenge to integrate into existing circuitry. Because of this particular feature, the olfactory system represents a unique opportunity to monitor axonal wiring and guidance, and to investigate synapse formation. Here we describe a procedure for in vivo labeling of sensory neurons and subsequent visualization of axons in the olfactory system of larvae of the amphibian Xenopus laevis. To stain sensory neurons in the olfactory organ we adopt the electroporation technique. In vivo electroporation is an established technique for delivering fluorophore-coupled dextrans or other macromolecules into living cells. Stained sensory neurons and their axonal processes can then be monitored in the living animal either using confocal laser-scanning or multiphoton microscopy. By reducing the number of labeled cells to few or single cells per animal, single axons can be tracked into the olfactory bulb and their morphological changes can be monitored over weeks by conducting series of in vivo time lapse imaging experiments. While the described protocol exemplifies the labeling and monitoring of olfactory sensory neurons, it can also be adopted to other cell types within the olfactory and other systems.

The Olfactory System as a Model to Study Axonal Growth Patterns and Morphology In Vivo

We describe a protocol for in vivo labeling of olfactory sensory neurons by electroporation and subsequent confocal laser-scanning or multiphoton microscopy to visualize neuronal morphology and its development over time.

모델로 후각 시스템은 축삭 성장 패턴 및 형태를 연구하는<em&gt; 생체</em

The olfactory system has the unusual capacity to generate new neurons throughout the lifetime of an organism. Olfactory stem cells in the basal portion of ...

<meta charset="utf8"></head><body>설치류 후각 실험을 위한 유연하고 비용 효율적인 후각계 설계

Constructing an Olfactometer for Rodent Olfactory Behavior Studies

The use of olfactometers to study rodent behavior and brain activity during olfactory tasks is crucial for understanding brain circuits. These sophisticated devices allow researchers to precisely control and deliver odor stimuli, enabling the investigation of complex olfactory processes in rodents. Although commercially available olfactometers are convenient, they present challenges when technical issues arise, often requiring costly assistance and potentially disrupting research timelines. This article details the construction of a custom olfactometer specifically designed for mouse olfactory behavior experiments, providing a comprehensive list of parts and step-by-step instructions. The olfactometer is controlled through MATLAB, offering a user-friendly interface for researchers. Importantly, the open-source code allows users to modify and adapt the system, tailoring behavioral tasks to meet specific experimental needs. Building a customized olfactometer empowers users with the knowledge and capability to perform custom experimental design and troubleshooting independently, saving both time and resources. This approach not only enhances research flexibility but also fosters a deeper understanding of the equipment's functionality, ultimately leading to more robust and reliable olfactory studies in rodents.

<meta charset="utf8"></head><body>설치류의 후각 행동 연구를 위한 후각계 구축, 근적외선 분광법, 심리 상담에서의 하이퍼스캐닝 연구

This protocol describes the construction of an olfactometer for go/no-go olfactory behavior experiments. Step-by-step instructions, along with images, are provided to ensure the successful construction of the olfactometer. Information for troubleshooting issues encountered during the process is also included.

설치류 후각 행동 연구를 위한 후각계 구축

The use of olfactometers to study rodent behavior and brain activity during olfactory tasks is crucial for understanding brain circuits. These ...

An Objective and Reproducible Test of Olfactory Learning and Discrimination in Mice

Olfaction is the predominant sensory modality in mice and influences many important behaviors, including foraging, predator detection, mating, and parenting. Importantly, mice can be trained to associate novel odors with specific behavioral responses to provide insight into olfactory circuit function. This protocol details the procedure for training mice on a Go/No-Go operant learning task. In this approach, mice are trained on hundreds of automated trials daily for 2&#8211;4 weeks and can then be tested on novel Go/No-Go odor pairs to assess olfactory discrimination, or be used for studies on how odor learning alters the structure or function of the olfactory circuit. Additionally, the mouse olfactory bulb (OB) features ongoing integration of adult-born neurons. Interestingly, olfactory learning increases both the survival and synaptic connections of these adult-born neurons. Therefore, this protocol can be combined with other biochemical, electrophysiological, and imaging techniques to study learning and activity-dependent factors that mediate neuronal survival and plasticity.

Here, we train mice on an associative learning task to test odor discrimination. This protocol also allows for studies on learning-induced structural changes in the brain.

신경 회로 토대 메모리 형성을 해부에 대한 신생아 흰쥐의 편측 냄새 학습 모델

요약

더 많은 비디오 탐색

이 비디오의 챕터

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

학습의 개별 단계에서 설치류에서 원격 음소거 신경 활동하는 방법

에 Appetitive 연관 강한 학습

초파리 성인 후각 충격 학습

A Low-Cost, Odor-Reward Association Task for Tests of Learning and Memory

모델로 후각 시스템은 축삭 성장 패턴 및 형태를 연구하는

설치류 후각 행동 연구를 위한 후각계 구축

설치류 후각 행동 연구를 위한 후각계 구축

설치류 후각 행동 연구를 위한 후각계 구축

후 각 학습 및 쥐에 있는 차별의 객관적이 고 재현성 시험