MSB is supported by the Alcohol Beverage Medical Research Foundation, a Center for Translational Research Award (UL1 TR000058), the National Institutes for Alcohol Abuse and Alcoholism (P50 AA022537), and startup funds provided by the Virginia Higher Education Equipment Trust Fund and the VCU School of Medicine.

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None of the authors have competing or conflicting interests.

여기에 제시된 방법은 마이크로 인젝션 캐뉼라 및 동기 동작의 분자 기판을 밝히는데 도움이 될 microinjectors를 제조하는 효율적인 수단이다. 이 방법은 몇 가지 장점을 제공한다. 첫째, 자신의 임플란트와 microinjectors을 제조하여, 새로운 실험 매개 변수를 신속하게 최적화 할 수있다, 즉, 하나가 도착하기를 맞춤 제작 한 부품을 기다릴 필요가 없습니다. 둘째, 정맥의 작은 직경으로 인해, 더 캐뉼라를 동시에 주입 할 수있다. 이것은 생존을 향상시킬 수 있습니다 필요한 수술 시간을 단축하고, 또한 동물 당 여러 개의 임플란트를 할 수 있습니다. 고정비 패러다임 빠르게 단순히 원하는 보강 일정 포함 된 이벤트 전이 파라미터리스트를 적용하여 프로그레시브 비율 패러다임으로 전환 될 수 있기 때문에, 셋째, 조건화 챔버를 제어하는​​데 사용되는 소프트웨어를 용이 프로그레시브 비 스케줄을 수용한다. 되려고광범위하게 유용한, 일반적인 미세 주입 절차는 현재 거의 모든 시약의 미세 주입을위한 광범위하게 적용해야한다고 제시했다. 따라서, 우리는이 기술은 약간의 수정과 향후 유사한 높은 유틸리티로 계속 될 것으로 예상하고있다. 단지 약간의 변수를 변경함으로써, 이러한 접근 방법은 시약의 다양한 개수에 적용될 수있다. 가장 일반적으로 조작 할 것이다 매개 변수는 마이크로 인젝터는 정맥 돌출 길이, 사출의 볼륨 및 주입 속도를 포함한다. 예를 들어, 하나의 주사기는 일반적으로 만성 임플란트 주위 아교 흉터 형성을 방지하기 위해 캐뉼라 팁에서 더 돌출 할 수있다. 또한, 하나의 더 큰 부피를 주입해도된다. 그 사용에 비하여 (10 분의 추가 시간 확산 자주 7 - - 10 분 및 3) 선조체 바이러스 microinjections 들어, 1 μL의 부피는 통상적으로 사용되며,이 볼륨은 전형적으로 장기간에 걸쳐 주입하여약리 시약 D (일반적으로 0.3 - 3 분 플러스 1 - - 2 이상 0.5 μL 3 분의 추가 확산 시간). 사용자는 문헌을 참조 및 / 또는 경험적으로 자신의 요구에 가장 적합한 매개 변수를 결정해야한다. 주사 전에 마이크로 인젝터의 분사 패턴의 1) 캐 뉼러의 길이, 2) 마이크로 인젝터 길이, 3)의 품질, 및 4) 시스템 무결성에 관계없이,이 절차의 성공은 4 변수 결정적으로 의존한다. 미세 주입 위치가 마이크로 인젝터는 정맥에서 돌출 깊이에 의존하기 때문에 모두 캐 뉼러 (단계 1.2.8) 및 마이크로 인젝터 길이 (포스트 굽힘, 단계 2.2.1)를 모두 정확하게 모든 주제 사이에 알려진 균일 한 것을,이 필수적이다 . 이것은 쉽게 용이라도 그 최종 재 측정에 필요한 길이 아니다 구현 거부에 의해 제어 될 수있다. 이 가이드 뉼러 바로 아래에 발생하는 경우 또한, 주입 위치 만 예측 될 수있다. 따라서, 어떤 마이크로 인젝터를 미상 그테스트에 긴, 미세 스트림을 뿌리지 S (단계 2.4.6)을 거부해야합니다. 품질 주입은 또한 종래 주입 시스템의 무결성에 관한 것이다. 여러 관광 명소 랩 닦아에서 관찰 된 인젝터에서 모든 물을 분배 후 (전 시약과 충전에) 경우, 누수가 (단계 2.4.8에 대한 참고) 해결 될 필요가있다. PE20 튜브 내의 물에서 약물을 분리 버블 (단계 2.4.9)가없는 한, 하나의 기포 (마이크로 인젝터 시약을 충전 한 후) 인 경우, 또한, 후 분사가 부분적으로 막혀있다. 이 방해하거나 예방 주사를 전환 할 수 중 하나. 이 너무 쉽게 해결 될 수있다 (참고 단계 2.4.8에서). 하나는 세 가지 대안이있는 동물이 고정대에있는 동안 microinject하고자합니다. 첫째, 하나는 정위 매니퓰레이터 의해 단단히 유지되고 또한 PE20 튜브에 연결을 허용하도록 충분히 연장 될 수 있도록 마이크로 인젝터 칼라의 길이를 증가시킬 수있다. 둘째,에전자는 일시적으로 정맥을 이식하고 여기에 제시된 표준 마이크로 인젝터를 사용할 수 있습니다. 셋째, 하나 그린과 광택 유리 피펫 사용할 수 있습니다. (16, 17) 여기에 제시된 방법의 중요한 한계는 최상의 절차에 익숙 잘 처리 래트에서 수행된다는 점이다. 동일한 조사자 2 개월 이상 매일 랫트를 취급하기 때문에 결과 절에 설명 된 데이터에 사용되는 래트 특별한 처리 절차가 필요 없다. 이것은 최소 2 주 동안 외과 임플란트의 매일 관찰 및 조작을 포함했다. 그러나, 래트 급격히 응력에 의해 영향을받을 수 프리 펄스 억제 분석 이전에 사용되는 다양한 기술에 의해 순응된다. 이러한 특수 습관화 기술들이 앞서 상세히 좋게되어있다. (43)이 절차 이외에, 그것은 쥐 단축 microinjectors가 뒤 사용되는 마이크로 인젝션 절차에 순응하는 것이 바람직하다반지 &#39;가짜&#39;주사. 이러한 허위 주사 동안, 마이크로 인젝터가 조직 손상을 제한하기 위하여 조직으로 돌출하지 않는 것이 중요하다. 즉, 마이크로 인젝터가 더 이상 14mm 이상 절곡되어서는 안된다. 따라서, 본 기술의 구현에 필요한 최적의 철저한 습관화는 제한으로 간주 될 수있다. 여러 행동 패러다임 동기를 측정하기 위해 존재하지만, 일반적으로는 비 진보적 피사체 보강제를 수득 발휘 기꺼이 노력을 정량화하는데 사용된다. 누진 비율 패러다임은 종종 최종 완료된 비율 레버 프레스의 최대 수로서 정의되는 브레이크 포인트로 알려진 측정 값을 만들어 낸다.. 즉, 최대 그 보강재를 생성 응답 21 프로그레시브 비가 크기 보강제 민감하다. 예를 들어, 높은 코카인 (또는 수 크로스) 투여 량은 더 높은 브레이크 코카인 및 하부 (또는 수 크로스) 복용 breakp 낮은 수율을 생산OINT. (21, 22) 따라서, 브레이크 포인트는 동기 부여 및 / 또는 보강 효과에 대한 일상적으로 사용하는 프록시입니다. 21,23-26 중단 점의 의도는 동물이 응답하지 않을 때 결정하기 때문에, 진보적 인 비율 패러다임의 중요한 매개 변수입니다 세션 길이. 세션 길이가 유한 한 브레이크 포인트는 거짓 값에 캡을 넣을 수 있으며, 이는 비정상적자가 투여 또는 증가 포스트 보강 일시의 속도를 감소 사전 치료에 의해 악화 될 수있다. .이 혼란은 접근 방법의 수에 의해 극복 될 수있다. 예를 들어, 세션 동물은 평균 간 투여 간격의 일부 복수로 응답을 원천 징수 한 경우 종료 (44)이 방법의 더 일반적으로 적용 변형이 응답 한 번 세션을 종료하는 것입니다 주제에 걸쳐 일정하게 유지된다 일부 경험적으로 결정된 값으로 원천 징수하고. 우리는 단계 4.4.9.11에이 방법을 적용 할 수있는 방법을 제공하고 있습니다.

Rodents and humans respond in remarkably similar ways to behaviorally relevant stimuli1-3. This suggests that rodents are appropriate subjects for elucidating the molecular substrates of behavior and complex psychiatric conditions4. Understanding the molecular substrates of complex behavioral processes, such as motivation, frequently requires brain microinjection. Both the brain microinjection technique and a primary motivation assay will be presented here. Rats will be used as subjects, but these procedures can readily be adapted to well-handled mice. Included herein are procedures for the manufacture of the required cannulae, obturators (dummy cannulae or stylets), and microinjectors. The method presented is significantly more flexible and more cost-efficient than prefabricated implements. This flexibility will prove valuable when optimizing conditions. Importantly, because the microinjection procedure can be used to test a myriad of hypotheses; the techniques presented here should be broadly applicable. For example, receptor ligands can be microinjected to understand neurochemistry3,5,6; cell-permeable peptides and small-molecules can be microinjected to understand intracellular signaling pathways7-10; toxins, ion channel blockers, or antagonist cocktails can be microinjected to understand circuitry1,11,12.
While the generic protocol presented here can be readily adapted by the user for their particular needs, the procedure is particularly well suited for behavioral assays since microinjection occurs in awake rodents that are only under mild hand restraint. No anesthesia or special restraints are required. This is possible because the brain itself lacks pain sensation. However, if anesthesia is not used, microinjection must occur through cannulae that were previously stereotaxically implanted. This is because nociceptors are present on the scalp, meninges,13 which are the membranes surrounding the brain, and the periosteum,14 which is the membrane covering the skull. It should be noted that microinjection under anesthesia is sometimes desirable. One example is when the virus is being injected, and one may wish to inject virus directly through either stainless steal needles15 or glass pipettes because this can reduce tissue damage and improve transduction efficiency.16,17 The microinjectors described below can be modified for this purpose and suggestions on how to do this can be found in the Discussion. Because other JoVE articles have demonstrated stereotaxic brain cannula implantation,18-20 these procedures will not be covered here.
We present these microinjection procedures together with an assay that quantifies motivation. Several rodent models of motivated behavior are currently in use, such as the runway box and barrier scaling. Here, we describe how to use an operant progressive ratio schedule of reinforcement to quantify motivation where operant responding is being maintained by a reinforcer. Responding on the progressive ratio is responsive to reinforcer magnitude.21,22 Accordingly, this assay is routinely used as a proxy for motivation and/or reinforcing efficacy. 21,23-30 Because several excellent reviews have covered this topic in detail,21,24 we will focus mainly on practical concerns.

<table border="0" cellpadding="0" cellspacing="0">
	<tbody>
		<tr>
			<td>Cannula Tubing</td>
			<td>Amazon Supply/ Small Parts</td>
			<td>HTXX-26T-60</td>
			<td>26 gauge, Hypotube S/S 316-TW 26GA</td>
		</tr>
		<tr>
			<td>Obturator</td>
			<td>Amazon Supply/ Small Parts</td>
			<td>GWXX-0080-30-05</td>
			<td>33 gauge, Wire S/S 316LVM 0.008 IN</td>
		</tr>
		<tr>
			<td>Microinjector Wire</td>
			<td>MicroGroup</td>
			<td>33RW 304</td>
			<td>33 gauge</td>
		</tr>
		<tr>
			<td>Super Glue</td>
			<td>Loctite</td>
			<td>3924AC</td>
			<td>Liquid, Non-gel, can be autoclaved</td>
		</tr>
		<tr>
			<td>Microinjector Plastic Tubing</td>
			<td>Becton Dickson</td>
			<td>427406</td>
			<td>PE20</td>
		</tr>
		<tr>
			<td>Medium Weight Hemostats</td>
			<td>World Precision Instruments</td>
			<td>501241-G</td>
			<td></td>
		</tr>
		<tr>
			<td>Ruler</td>
			<td>Fisher</td>
			<td>09-016</td>
			<td>150 mm</td>
		</tr>
		<tr>
			<td>#7 Forceps</td>
			<td>Stoelting</td>
			<td>52100-77</td>
			<td>Dumont, Dumostar</td>
		</tr>
		<tr>
			<td>Rotary Tool</td>
			<td>Dremmel</td>
			<td>285</td>
			<td>Two-speeds</td>
		</tr>
		<tr>
			<td>Cut-off Disc</td>
			<td>McMaster Carr</td>
			<td>3602</td>
			<td>15/16&#34; x 0.025&#34;</td>
		</tr>
		<tr>
			<td>Microinjection Pump</td>
			<td>Harvard Apparatus</td>
			<td>PhD 2000</td>
			<td></td>
		</tr>
		<tr>
			<td>1 ul Glass Syringe</td>
			<td>Hamilton</td>
			<td>7001KH</td>
			<td>Needle Style: 25s/2.75&#34;/3</td>
		</tr>
		<tr>
			<td>Cotton Tipped Applicator</td>
			<td>Fisher</td>
			<td>23-400-101</td>
			<td></td>
		</tr>
		<tr>
			<td>Lab Wipes</td>
			<td>Kimwipes</td>
			<td>34133</td>
			<td></td>
		</tr>
		<tr>
			<td>Operant Software</td>
			<td>Coulbourn</td>
			<td>Graphic State</td>
			<td></td>
		</tr>
		<tr>
			<td>Operant Chambers</td>
			<td>Coulborun</td>
			<td>Habitest</td>
			<td></td>
		</tr>
	</tbody>
</table>

rodent brain microinjection to study molecular substrates of motivated behavior

Brain microinjection can aid elucidation of the molecular substrates of complex behaviors, such as motivation. For this purpose rodents can serve as appropriate models, partly because the response to behaviorally relevant stimuli and the circuitry parsing stimulus-action outcomes is astonishingly similar between humans and rodents. In studying molecular substrates of complex behaviors, the microinjection of reagents that modify, augment, or silence specific systems is an invaluable technique. However, it is crucial that the microinjection site is precisely targeted in order to aid interpretation of the results. We present a method for the manufacture of surgical implements and microinjection needles that enables accurate microinjection and unlimited customizability with minimal cost. Importantly, this technique can be successfully completed in awake rodents if conducted in conjunction with other JoVE articles that covered requisite surgical procedures. Additionally, there are many behavioral paradigms that are well suited for measuring motivation. The progressive ratio is a commonly used method that quantifies the efficacy of a reinforcer to maintain responding despite an (often exponentially) increasing work requirement. This assay is sensitive to reinforcer magnitude and pharmacological manipulations, which allows reinforcing efficacy and/ or motivation to be determined. We also present a straightforward approach to program operant software to accommodate a progressive ratio reinforcement schedule.

여기에서 우리는 상기 절차를 얻을 수있다 결과의 예를 도시한다. 표시된 첫 번째는 바이러스 벡터 (그림 3)에 의해 형질 전환 된 전형적인 지역이다. 일반적으로 3 ~ 1mm의 형질 볼륨 선조체 조직을 얻을 수있다. 형질 영역의 부피 카발리의 방법을 사용하여 직렬 섹션에 걸쳐 정량화 될 수있다 (32)이 중요한 것은, 형질 볼륨은 형질되는 조직의 형태와 같은 많은 요인에 의존한다.; 바이러스 혈청 형; 프로모터; 속도 및 주입 부피; 주입 된 입자의 수​​; 인젝 산도; 및 만니톨과 같은 삼투압 시약, 여부, 사용되었다. 33, 34 일반적으로, 우리는 10 분에 걸쳐 10 ~ 12 입자 / ㎖, 1 μL /면을 microinject 및 obturators 교체 7 분 추가 이전 할 수 있습니다. 또한, pH가 테이트 8. 다음 주위 통상, 우리는 동기가 어느 유전자의 발현에 의해 조작 될 수 있음을 보여준다 (도 4)또는 약리 시약 미세 주입 (그림 5). 그림 4에서 우리는 독점적으로 디자이너 마약 (DREADDs)에 의해 활성화 디자이너 수용체를 표명했다. DREADD 수용체 코딩 영역은 내부 리보솜 항목 사이트 및 mCitrine 카세트으로 이어졌습니다. mCitrine는 형질 감염된 세포의 편리를 육안으로 확인 할 수 있습니다. DREADD는 이종삼 G- 단백질 Gαq에 커플 링시켰다. 성상 세포, 28,35 및 DREADD 자체를 자극 할 수 Gαq 결합 DREADD의 활성화는 클로자핀-N-산화물 (CNO, 3 ㎎ / ㎏, IP)의 전신 투여에 의해 활성화 될 수있다. 14,22,36 쥐에 훈련을받은 레버는 3 레버 프레스 60 연속 일 동안 매일 1 시간 세션 동안 마실 1 기회를 얻었다 에탄올 보강에 대한 반응한다. 다음, 쥐 금욕을 강요하고, 핵 핵심 성상 세포를 중격 의지에 Gαq 결합 DREADDs 표현했다. 삼주의 절제, 자기 ADMI에 의욕 후<html> nister 에탄올 브레이크 포인트에 의해 측정 하였다. 핵 21,27-30 활성화 CNO 전신 투여를 통해, 절제는 차량에 비해 후 에탄올자가 투여를 재개 쥐의 의욕을 감소 코어 성상 중격 의지. 중요한 것은, CNO 대신 DREADD의 녹색 형광 단백질을 발현시켰다 동등 훈련 코호트에 영향을 미치지 않았다. <img alt="그림 3" src="/files/ftp_upload/53018/53018fig3.jpg" /> 마이크로 인젝션 된 바이러스에 의해 형질도 3 대표 복측 선조체 지역.이 이미지는 핵의 영역은 위의 방법에 따라 형질 감염시켰다 아스트로 중격 의지를 나타낸다. 데이터는 저작권 소유자의 허가를 불 등. 13에서 재 인쇄되어 있습니다. 그 세부 사항은 첨부 된 간행물 및 보충 자료에서 발견 될 수있다.&quot;빈&gt;이 그림의 더 큰 버전을 보려면 여기를 클릭하십시오. <img alt="그림 4" src="/files/ftp_upload/53018/53018fig4.jpg" /> 자기 관리] 에탄올에 쥐 그림 4. 동기 부여는 핵 핵심 성상 세포를 중격 의지에. 표현을 통해-바이러스가 표현 할 일주를 허용 한 후 바이러스가 미세 주입하고, 동기는 브레이크 포인트를 통해 측정 한 형질 전환 유전자를 활성화하여 감소 하였다. 유전자는 전용 디자이너 마약 (DREADDs)에 의해 디자이너 수용체 활성화 된 표현. 일원 분산 분석 (F (2,30) = 3.29, P = 0.04)은 Scheffe 사후에 의해 밝혀 다음에 그 클로자핀-N-산화물의 전신 투여에 의한 DREADD의 활성화 (CNO, 3 ㎎ / ㎏, IP ) 크게 대신 DR의) CNO는 녹색 형광 단백질 (GFP를 표현했다 동등하게 교육을받은 집단에 영향을 미치지 아니합니다 차량에 비해 절제 후자가 관리] 에탄올에 쥐의 의욕을 감소EADD. 데이터는 저작권 소유자의 허가를 불 등. 13에서 재 인쇄되어 있습니다. 그 세부 사항은 첨부 된 간행물 및 보충 자료에서 발견 될 수있다. <img alt="그림 5" src="/files/ftp_upload/53018/53018fig5.jpg" /> 그림 간극 결합 차단제 5. 미세 주입은 절제 후 에탄올을-관리 자체에 쥐의 의욕을 증가 두 간극 결합 차단제는 에탄올 관리 자체에 쥐 (브레이크 포인트를 통해 측정) 동기 부여에 미치는 영향을 평가 하였다 :. 메플로퀸 및 18 α - 글리시 레틴 산 (18-α)를. 두 일원 분산 분석은 핵으로 갭 접합 차단제의 미세 주사가 핵심이 절제 후 에탄올을-관리 자체에 쥐의 의욕을 증가 중격 의지 것으로 나타났다 (F (3,40) = 5.56, P = 0.003). 데이터는 저작권 소유자의 허가를 불 등. 13에서 재 인쇄되어 있습니다. 자세한 사항은이 책에서 찾을 수 있으며첨부 된 보충 자료.

Watch this Scientific Journal Video about Rodent Brain Microinjection to Study Molecular Substrates of Motivated Behavior at JoVE.com

Rodent Brain Microinjection to Study Molecular Substrates of Motivated Behavior

Rodents are an appropriate model to investigate the molecular substrates of behavior and complex psychiatric disorders. Brain microinjection in awake rodents can be used to elucidate disease substrates. An efficient and customizable brain microinjection method as well as the execution of an operant paradigm that quantifies motivation is presented.

쥐 뇌 미세 주입은 분자 기판 동기를 행동을 연구하기 위해

Brain microinjection can aid elucidation of the molecular substrates of complex behaviors, such as motivation. For this purpose rodents can serve as ...

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Behavior

14.2K 조회수.  Virginia Commonwealth University School of Medicine. Rodents are an appropriate model to investigate the molecular substrates of behavior and complex psychiatric disorders. Brain microinjection in awake rodents can be used to elucidate disease substrates. An efficient and customizable brain microinjection method as well as the execution of an operant paradigm that quantifies motivation is presented. 

비디오: Rodent Brain Microinjection to Study Molecular Substrates of Motivated Behavior

Stereotaxic Microinjection of Viral Vectors Expressing Cre Recombinase to Study the Role of Target Genes in Cocaine Conditioned Place Preference

Microinjecting recombinant adenoassociated viral (rAAV) vectors expressing Cre recombinase into distinct mouse brain regions to selectively knockout genes of interest allows for enhanced temporally- and regionally-specific control of gene deletion, compared to existing methods. While conditional deletion can also be achieved by mating mice that express Cre recombinase under the control of specific gene promoters with mice carrying a floxed gene, stereotaxic microinjection allows for targeting of discrete brain areas at experimenter-determined time points of interest. In the context of cocaine conditioned place preference, and other cocaine behavioral paradigms such as self-administration or psychomotor sensitization that can involve withdrawal, extinction and/or reinstatement phases, this technique is particularly useful in exploring the unique contribution of target genes to these distinct phases of behavioral models of cocaine-induced plasticity. Specifically, this technique allows for selective ablation of target genes during discrete phases of a behavior to test their contribution to the behavior across time. Ultimately, this understanding allows for more targeted therapeutics that are best able to address the most potent risk factors that present themselves during each phase of addictive behavior.

This article describes how to microinject viral vectors into mouse brain and then test in a conditioned place preference paradigm that includes an acquisition, extinction and reinstatement phase.

크레 재조합이 코카인 컨디셔닝 장소 환경에있는 표적 유전자의 역할을 연구하는 표현 바이러스 성 벡터의 정위 미세 주입

Microinjecting recombinant adenoassociated viral (rAAV) vectors expressing Cre recombinase into distinct mouse brain regions to selectively knockout genes ...

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Uncovering Beat Deafness: Detecting Rhythm Disorders with Synchronized Finger Tapping and Perceptual Timing Tasks

A set of behavioral tasks for assessing perceptual and sensorimotor timing abilities in the general population (i.e., non-musicians) is presented here with the goal of uncovering rhythm disorders, such as beat deafness. Beat deafness is characterized by poor performance in perceiving durations in auditory rhythmic patterns or poor synchronization of movement with auditory rhythms (e.g., with musical beats). These tasks include the synchronization of finger tapping to the beat of simple and complex auditory stimuli and the detection of rhythmic irregularities (anisochrony detection task) embedded in the same stimuli. These tests, which are easy to administer, include an assessment of both perceptual and sensorimotor timing abilities under different conditions (e.g., beat rates and types of auditory material) and are based on the same auditory stimuli, ranging from a simple metronome to a complex musical excerpt. The analysis of synchronized tapping data is performed with circular statistics, which provide reliable measures of synchronization accuracy (e.g., the difference between the timing of the taps and the timing of the pacing stimuli) and consistency. Circular statistics on tapping data are particularly well-suited for detecting individual differences in the general population. Synchronized tapping and anisochrony detection are sensitive measures for identifying profiles of rhythm disorders and have been used with success to uncover cases of poor synchronization with spared perceptual timing. This systematic assessment of perceptual and sensorimotor timing can be extended to populations of patients with brain damage, neurodegenerative diseases (e.g., Parkinson&#8217;s disease), and developmental disorders (e.g., Attention Deficit Hyperactivity Disorder).

Behavioral tasks that allow for the assessment of perceptual and sensorimotor timing abilities in the general population (i.e., non-musicians) are presented. Synchronization of finger tapping to the beat of an auditory stimuli and detecting rhythmic irregularities provides a means of uncovering rhythm disorders.

비트의 청각 장애를 폭로 : 감지 리듬 장애를 누르면 동기화 손가락 및 지각 타이밍 작업에

A set of behavioral tasks for assessing perceptual and sensorimotor timing abilities in the general population (i.e., non-musicians) is presented here ...

A Dual Task Procedure Combined with Rapid Serial Visual Presentation to Test Attentional Blink for Nontargets

When viewers search for targets in a rapid serial visual presentation (RSVP) stream, if two targets are presented within about 500 msec of each other, the first target may be easy to spot but the second is likely to be missed. This phenomenon of attentional blink (AB) has been widely studied to probe the temporal capacity of attention for detecting visual targets. However, with the typical procedure of AB experiments, it is not possible to examine how the processing of non-target items in RSVP may be affected by attention. This paper describes a novel dual task procedure combined with RSVP to test effects of AB for nontargets at varied stimulus onset asynchronies (SOAs). In an exemplar experiment, a target category was first displayed, followed by a sequence of 8 nouns. If one of the nouns belonged to the target category, participants would respond &#8216;yes&#8217; at the end of the sequence, otherwise participants would respond &#8216;no&#8217;. Two 2-alternative forced choice memory tasks followed the response to determine if participants remembered the words immediately before or after the target, as well as a random word from another part of the sequence. In a second exemplar experiment, the same design was used, except that 1) the memory task was counterbalanced into two groups with SOAs of either 120 or 240 msec and 2) three memory tasks followed the sequence and tested remembrance for nontarget nouns in the sequence that could be anywhere from 3 items prior the target noun position to 3 items following the target noun position. Representative results from a previously published study demonstrate that our procedure can be used to examine divergent effects of attention that not only enhance targets but also suppress nontargets. Here we show results from a representative participant that replicated the previous finding.&#160;

This paper describes a novel dual task procedure combined with Rapid Serial Visual Presentation to look at attentional blink at varied stimulus onset asynchronies. This experimental procedure differed from others in that it tested attentional blink for nontargets.

Nontargets에 대한주의 집중 깜박임을 테스트하는 빠른 직렬 비주얼 프리젠 테이션과 결합하여 이중 작업 절차

When viewers search for targets in a rapid serial visual presentation (RSVP) stream, if two targets are presented within about 500 msec of each other, the ...

Using the Activity-based Anorexia Rodent Model to Study the Neurobiological Basis of Anorexia Nervosa

Anorexia nervosa (AN) is a psychiatric illness characterized by excessively restricted caloric intake and abnormally high levels of physical activity. A challenging illness to treat, due to the lack of understanding of the underlying neurobiology, AN has the highest mortality rate among psychiatric illnesses. To address this need, neuroscientists are using an animal model to study how neural circuits may contribute toward vulnerability to AN and may be affected by AN. Activity-based anorexia (ABA) is a bio-behavioral phenomenon described in rodents that models the key symptoms of anorexia nervosa. When rodents with free access to voluntary exercise on a running wheel experience food restriction, they become hyperactive &#8211; running more than animals with free access to food. Here, we describe the procedures by which ABA is induced in adolescent female C57BL/6 mice. On postnatal day 36 (P36), the animal is housed with access to voluntary exercise on a running wheel. After 4 days of acclimation to the running wheel, on P40, all food is removed from the cage. For the next 3 days, food is returned to the cage (allowing animals free food access) for 2 hr daily. After the fourth day of food restriction, free access to food is returned and the running wheel is removed from the cage to allow the animals to recover. Continuous multi-day analysis of running wheel activity shows that mice become hyperactive within 24 hr following the onset of food restriction. The mice run even during the limited time during which they have access to food. Additionally, the circadian pattern of wheel running becomes disrupted by the experience of food restriction. We have been able to correlate neurobiological changes with various aspects of the animals&#8217; wheel running behavior to implicate particular brain regions and neurochemical changes with resilience and vulnerability to food-restriction induced hyperactivity.

Here we present a protocol to induce activity-based anorexia (ABA) in female adolescent mice. ABA is a condition of hyperactivity evoked by imposing food restriction on rodents with access to a running wheel. This phenomenon is being used as a model to study the underlying neurobiology of anorexia nervosa.

활동 기반 식욕 부진 설치류 모델을 사용하면 거식증의 신경 생물학적 기초 연구에

Anorexia nervosa (AN) is a psychiatric illness characterized by excessively restricted caloric intake and abnormally high levels of physical activity. A ...

Combined Shuttle-Box Training with Electrophysiological Cortex Recording and Stimulation as a Tool to Study Perception and Learning

Shuttle-box avoidance learning is a well-established method in behavioral neuroscience and experimental setups were traditionally custom-made; the necessary equipment is now available by several commercial companies. This protocol provides a detailed description of a two-way shuttle-box avoidance learning paradigm in rodents (here Mongolian gerbils; Meriones unguiculatus) in combination with site-specific electrical intracortical microstimulation (ICMS) and simultaneous chronical electrophysiological in vivo recordings. The detailed protocol is applicable to study multiple aspects of learning behavior and perception in different rodent species.
Site-specific ICMS of auditory cortical circuits as conditioned stimuli here is used as a tool to test the perceptual relevance of specific afferent, efferent and intracortical connections. Distinct activation patterns can be evoked by using different stimulation electrode arrays for local, layer-dependent ICMS or distant ICMS sites. Utilizing behavioral signal detection analysis it can be determined which stimulation strategy is most effective for eliciting a behaviorally detectable and salient signal. Further, parallel multichannel-recordings using different electrode designs (surface electrodes, depth electrodes, etc.) allow for investigating neuronal observables over the time course of such learning processes. It will be discussed how changes of the behavioral design can increase the cognitive complexity (e.g. detection, discrimination, reversal learning).

Shuttle-box avoidance learning is well-established in behavioral neuroscience. This protocol describes how shuttle-box learning in rodents can be combined with site-specific electrical intracortical microstimulation (ICMS) and simultaneous chronical in vivo recordings as a tool to study multiple aspects of learning and perception.

인식 및 학습을 연구하는 도구로 전기 생리 텍스 녹화 및 자극과 셔틀 박스 교육을 결합

Shuttle-box avoidance learning is a well-established method in behavioral neuroscience and experimental setups were traditionally custom-made; the ...

The Rodent Psychomotor Vigilance Test (rPVT): A Method for Assessing Neurobehavioral Performance in Rats and Mice

The human Psychomotor Vigilance Test (PVT) is a widely used procedure for measuring changes in fatigue and sustained attention. The present article describes a rodent version of the PVT&#8212;termed the &#34;rPVT&#34;&#8212;that measures similar aspects of attention (i.e., performance accuracy, motor speed, premature responding, and lapses in attention). Data are presented that demonstrate both the short- and long-term usefulness of the rPVT when employed with laboratory rats. Rats easily learn the rPVT, and learning to perform the basic procedure takes less than two weeks of training. Once acquired, rat performances in the rPVT show a high degree of similarity to these same performance measures in the human PVT, including similarities in, lapses in attention, reaction times, vigilance decrements across session time (i.e., the human &#34;time-on-task&#34; effects), and the response-stimulus interval (RSI) effect described for humans. Thus the rPVT can be an extremely valuable tool for assessing the effects of a wide range of variables on sustained attention quite similar to human PVT performances, and thus can be useful for developing novel treatments for neurobehavioral dysfunctions.

The Rodent Psychomotor Vigilance Test rPVT: A Method for Assessing Neurobehavioral Performance in Rats and Mice

A rat version of the human Psychomotor Vigilance Test (PVT) is described that measures aspects of attention similar to those measured with the human PVT, including aspects of human vigilance such as performance accuracy, motor speed, premature responding, and lapses in attention.

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

The human Psychomotor Vigilance Test (PVT) is a widely used procedure for measuring changes in fatigue and sustained attention. The present article ...

Reversible Cooling-induced Deactivations to Study Cortical Contributions to Obstacle Memory in the Walking Cat

On complex, naturalistic terrain, sensory information about an environmental obstacle can be used to rapidly adjust locomotor movements for avoidance. For example, in the cat, visual information about an impending obstacle can modulate stepping for avoidance. Locomotor adaptation can also occur independent of vision, as sudden tactile inputs to the leg by an expected obstacle can modify the stepping of all four legs for avoidance. Such complex locomotor coordination involves supraspinal structures, such as the parietal cortex. This protocol describes the use of reversible, cooling-induced cortical deactivation to assess parietal cortex contributions to memory-guided obstacle locomotion in the cat. Small cooling loops, known as cryoloops, are specially shaped to deactivate discrete regions of interest to assess their contributions to an overt behavior. Such methods have been used to elucidate the role of parietal area 5 in memory-guided obstacle avoidance in the cat.

Complex locomotion in naturalistic environments requiring careful coordination of the limbs involves regions of the parietal cortex. The following protocol describes the use of reversible cooling-induced deactivation to demonstrate the role of parietal area 5 in memory-guided obstacle avoidance in the walking cat.

가역 냉각 유도 Deactivations 공부 장애물 메모리 걷는 고양이에 대뇌 피 질의 기여를

On complex, naturalistic terrain, sensory information about an environmental obstacle can be used to rapidly adjust locomotor movements for avoidance. For ...

Central and Divided Visual Field Presentation of Emotional Images to Measure Hemispheric Differences in Motivated Attention

Two dominant theories on lateralized processing of emotional information exist in the literature. One theory posits that unpleasant emotions are processed by right frontal regions, while pleasant emotions are processed by left frontal regions. The other theory posits that the right hemisphere is more specialized for the processing of emotional information overall, particularly in posterior regions.
Assessing the different roles of the cerebral hemispheres in processing emotional information can be difficult without the use of neuroimaging methodologies, which are not accessible or affordable to all scientists. Divided visual field presentation of stimuli can allow for the investigation of lateralized processing of information without the use of neuroimaging technology. 
This study compared central versus divided visual field presentations of emotional images to assess differences in motivated attention between the two hemispheres. The late positive potential (LPP) was recorded using electroencephalography (EEG) and event-related potentials (ERPs) methodologies to assess motivated attention. Future work will pair this paradigm with a more active behavioral task to explore the behavioral impacts on the attentional differences found.

This study compared central versus divided visual field presentations of emotional images to assess differences in motivated attention between the two hemispheres. The late positive potential (LPP) was recorded using electroencephalography (EEG) and event-related potentials (ERPs) methodologies to assess motivated attention.

시야의 중앙와 프레 젠 테이 션의 정서적 이미지 동기 관심에 반구형 차이 측정

Two dominant theories on lateralized processing of emotional information exist in the literature. One theory posits that unpleasant emotions are processed ...

Combining Quantitative Food-intake Assays and Forcibly Activating Neurons to Study Appetite in Drosophila

Food consumption is under the tight control of the brain, which integrates the physiological status, palatability, and nutritional contents of the food, and issues commands to start or stop feeding. Deciphering the processes underlying the decision-making of timely and moderate feeding carries major implications in our understanding of physiological and psychological disorders related to feeding control. Simple, quantitative, and robust methods are required to measure the food ingestion of animals after experimental manipulation, such as forcibly increasing the activities of certain target neurons. Here, we introduced dye-labeling-based feeding assays to facilitate the neurogenetic study of feeding control in adult fruit flies. We review available feeding assays, and then describe our methods step-by-step from setup to analysis, which combine thermogenetic and optogenetic manipulation of neurons controlling feeding motivation with dye-labeled food intake assay. We also discuss the advantages and limitations of our methods, compared with other feeding assays, to help readers choose an appropriate assay.

Combining Quantitative Food-intake Assays and Forcibly Activating Neurons to Study Appetite in Drosophila

Quantitative food-intake assays with dyed food provide a robust and high-throughput means to evaluate feeding motivation. Combining the food consumption assay with thermogenetic and optogenetic screens is a powerful approach to investigate the neural circuits underlying appetite in adult Drosophila melanogaster.

양이 많은 음식 섭취 량 분석을 결합 하 고 강제로 초파리 에서 식욕을 공부 하는 뉴런을 활성화

Food consumption is under the tight control of the brain, which integrates the physiological status, palatability, and nutritional contents of the food, ...

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 ...