Name: assessment of cocaine-induced behavioral sensitization and conditioned place preference in mice
Uploaded: 2016-02-18T13:00:00
Description: Watch this Scientific Journal Video about Assessment of Cocaine-induced Behavioral Sensitization and Conditioned Place Preference in Mice at JoVE.com

저자는 행동 테스트에 대한 도움말은 카렌 디 에츠와 샤리 번바움 행동 설계 고려 사항에 이전 입력 및 로렌 PECA 감사합니다. 저자는 또한 시몬스 재단 (CWC에 시몬스 재단 자폐증 연구 이니셔티브 부여), NIDA (DA008277, DA027664 및 DA030590 CWC, RDP에 LNS에 F32DA027265 및 F32DA036319에), FRAXA 연구 재단과 엘레과 마일의 관대 한 지원을 인정 해안 원정대 프로그램 (LNS에 교제 지원), 그리고 존 Kaneb 원정대 프로그램 (MT에 교제 지원).

모든 실험 절차는 맥린 병원 기관 동물 관리 및 사용위원회에 의해 승인되었습니다. 참고 : 다음 프로토콜은 다른 성공적인 프로토콜 (예를 들어, 라이트 - 대 어두운 상 시험, 연속 대 간헐적 투여 등)과 다른 많은 세부있는 CPP와 전위의 민감성에 하나의 접근 방법을 설명합니다. 초보자는 손에서 실험 질문 (들)을 기반으로 문헌에서 변화 적응, 이러한 프로토콜로 시작하거나 가이드로 사용...

이 프로토콜은 약물 유발 행동 가소성의 양상을 평가하기 위해 평균 랩에 의해 사용될 수있는 각각의 조건화 된 장소 선호도 전위의 증감에 대한 방법을 설명한다. 대부분의 행동 테스트와 마찬가지로, 기본 프로토콜 이외의 추가 가치 고려 사항이 있습니다. 우선, 이들 기술들 각각은 두 단계, 유도 식을 갖는 것으로 생각할 수있다. &quot;유도&quot;행동을 위해 컨디셔닝 동안 발생 CPP의 개발을 포?...

Research aimed at understanding drug addiction using animal models must take a variety of approaches to address each of the assorted components that obstruct treatment success, including reward/reinforcement/motivation and withdrawal and relapse, as well as the general persistence that further complicates these issues in addiction. Since rewarding experiences associated with taking a drug of abuse are thought to motivate subsequent use, studies focusing on drug-context associations may be particularly useful for understanding brain mechanisms that contribute to drug taking and seeking. One such assay, conditioned place preference (CPP) is a high-throughput method for comparing group differences in reward sensitivity. The traditional interpretation of the task involves classical, or Pavlovian, conditioning, where a conditioned stimulus (CS) is paired with an unconditioned stimulus (UCS), and after multiple pairings, the CS elicits the same behavior as the UCS (however, see39,40). Theoretically, animals learn to associate an interoceptive state (reward or aversion) with contextual cues. The relative aversive or appetitive intensity of the interoceptive state is then assessed by then determining the animal's preference for the contextual cues. The use of place conditioning to measure drug-reward associations dates back to at least 1957, to a study using morphine on rats in a Y-maze3,4. Over the past several decades, variations on this method have been widely used to study place preference and aversion in rodents to various stimuli, and it remains particularly useful in the study of associations induced by drugs of abuse. In drug-addiction research, the assay has been used to assess the rewarding properties of a number of drugs and the contribution of different brain systems and proteins to drug reward (for reviews, see5-7,44). While there are superior methods of assessing factors that contribute to drug addiction, namely drug self-administration, CPP is a simple and much more accessible approach to measuring reward function. 
Most current protocols for conditioned place preference and aversion (CPA) use an apparatus that allows rodents to have access to two distinct chambers, either via a doorway or smaller connecting chamber. Distinctions between the two chambers are often based, at a minimum, on visual and tactile cues, including wall color and floor texture, but sometimes include other elements, such as olfactory cues. "Biased" designs typically attempt to reverse a pre-existing, innate preference for one chamber over the other, such as the one that rodents generally show for a black chamber over white. "Unbiased" designs aim to create a preference to one of two chambers that were initially equally appealing by randomly counterbalancing assignment to either chamber within a group. A "balanced" design is used when animals show small preferences, but do not, as a group, favor the same chamber. Goals of this latter design are to produce 1) pre-test preference scores for the (eventual) cocaine-paired chamber that are not significantly different between experimental groups and 2) negligible preference for the cocaine-paired chamber at pre-test, either positive or negative8. The balanced design is ideal for use with the described chambers, which utilize contradicting biases for wall color (black over white) and flooring (wire over bar), resulting in a roughly equal distribution of small preferences for both the black and white sides in different animals. Balancing calculations are described in further detail below.
During conditioning, animals are exposed to a drug and quickly placed into one of these two environments for a limited time period. Exposure is typically via intraperitoneal (i.p.) or sometimes subcutaneous (s.c.) injection, although paradigms for intravenous (i.v.) self-administration9, and intracranial infusions38 in a place preference apparatus have also been developed. These pairings are complemented by non-drug (vehicle) pairings of the same length conducted in the opposite chamber, which can take place on the same day as drug pairings or on separate days. In general, when allowed to explore the apparatus after conditioning, animals will spend more time where they received a rewarding drug (i.e., one that humans and animals will voluntarily self-administer), while they will avoid a place where they were given a drug that induced illness (e.g., lithium chloride). Several studies have been dedicated to optimizing the conditions for place preference to different drugs of abuse (for review, see7). Cocaine doses (i.p.) for mice generally range from 1 to 20 mg/kg, with doses less than 5 mg/kg often used to parse high sensitivity in one group. Two or more drug pairings are typically required for adult mice10, and the length of these pairings is an important consideration. Very low doses of cocaine require an immediate and brief conditioning, likely because this method captures the most rewarding period of the exposure. Delayed or very long conditioning periods can result in no preference, or may even induce aversion11,12. Here is presented a basic method for obtaining conditioned place preference to cocaine in adult mice.
While the CPP assay is an ideal method for assessing reward-related learning and memory of drug-context associations, behavioral sensitization is arguably easier to perform and allows the assessment of changes that develop over repeated treatment. Also known as reverse-tolerance, behaviors undergoing sensitization are incrementally enhanced over repeated exposures to a particular drug of abuse, especially psychostimulants, and cross-sensitization is known to occur between some, but not all, of these drugs. One of the first assessments of cocaine-induced locomotor sensitization, in particular, in rodents was published in 197613. A number of labs have shown that sensitized locomotion is detectable long after drug cessation, depending on the original length, location and dose of exposure14-17, and the current protocol has been used to detect sensitization as long as 10 months following seven days (30 mg/kg) of cocaine treatment in mice18. The test can be performed using either photobeam or video-tracking technologies, in apparatuses of differing sizes and shapes, making it simple for many labs to perform. The robust nature, simplicity and persistence of locomotor sensitization makes its assessment an ideal part of examining basic mechanisms of long-lasting changes in drug-induced behavior. 
As is expanded upon in the discussion, an important consideration when performing the locomotor sensitization assay is whether drug is given in the home- or test-cage environment. To take advantage of the robust sensitization that occurs when drug administration occurs outside of the home cage, this protocol employs this method. However, it has been observed that when animals are not adequately habituated to a new environment before drug exposure, a novelty-induced ceiling effect occurs on Day 1, which can partially or fully mask the progressive nature of sensitization. It is likely that this represents synergistic locomotor-activating effects of the drug together with novelty, and while the mechanisms underlying such effects may be interesting, the method described is designed to reduce the role of novelty and allow the effects of the drug to be measured more independently. While it is expected this method will be useful in the assessment of other locomotor-sensitizing drugs, it has primarily evaluated its effectiveness with cocaine in C57BL/6 mice.

<table border="0" cellpadding="0" cellspacing="0" width="1293">
	<tbody>
		<tr height="42">
			<td height="42" style="height:42px;width:215px;">Cocaine Hydrochloride USP</td>
			<td style="width:127px;">Mallinckrodt Pharmaceuticals</td>
			<td style="width:119px;">0406-1520</td>
			<td style="width:832px;">Purchase and use (Schedule II controlled substance) for research purposes requires compliance and licensure according to state and federal law.&#160;</td>
		</tr>
		<tr height="84">
			<td height="84" style="height:84px;width:215px;">Conditioned Place Preference,&#160; Three Compartment Apparatus with Manual Doors and Lights for Mouse</td>
			<td style="width:127px;">Med-Associates Inc.</td>
			<td style="width:119px;">MED-CPP-MS &#38; MED-CPP-3013</td>
			<td>Our laboratory has used these boxes; however, many alternative boxes are available &#38; acceptable.</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:215px;">PAS-Home Cage Activity Monitoring Photobeam Arrays</td>
			<td style="width:127px;">San Diego Instruments</td>
			<td style="width:119px;">2325-0223 &#38; 7500-0221</td>
			<td>Our lab houses these arrays inside of custom built chambers, as described in the text.&#160; There are alternatives available.</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:215px;">Disposable Sani-Cloth disenfecting wipes</td>
			<td style="width:127px;">PDI</td>
			<td style="width:119px;">13872</td>
			<td></td>
		</tr>
	</tbody>
</table>

assessment of cocaine-induced behavioral sensitization and conditioned place preference in mice

It is thought that rewarding experiences with drugs create strong contextual associations and encourage repeated intake. In turn, repeated exposures to drugs of abuse make lasting alterations in the brain function of vulnerable individuals, and these persistent alterations likely serve to maintain the maladaptive drug seeking and taking behaviors characteristic of addiction/dependence2. In rodents, reward experience and contextual associations are frequently measured using the conditioned place preference assay, or CPP, wherein preference for a previously drug-paired context is measured. Behavioral sensitization, on the other hand, is an increase in a drug-induced behavior that develops progressively over repeated exposures. Since sensitized behaviors can often be measured after several months of drug abstinence, depending on the dose and length of initial exposure, they are considered observable correlates of lasting drug-induced plasticity. Researchers have found these assays useful in determining the neurobiological substrates mediating aspects of addiction as well as assessing the potential of different interventions in disrupting these behaviors. This manuscript describes basic, effective protocols for mouse CPP and locomotor behavioral sensitization to cocaine.

CPP 분석의 대표적인 결과는 대략 세의 구주를 야생형 C57BL / 6N 마우스를 사용하여 그림 6에 표시됩니다. 연구 설계 시험 (전후)의 내 - 과목 변수로, 2 × 3 혼합 요인이었고,이 치료 (생리 식염수와 코카인 5, 10 ㎎ / ㎏)의 변수를 사이에-과목. RM ANOVA 두 시험에서 관찰 유의 한 주 효과 대신에 해석 된 테스트 및 치료 (F 2,20 = 3.68, P &lt;0.05) 사이에 유의 한 상호 작용을 보여 주?...

Watch this Scientific Journal Video about Assessment of Cocaine-induced Behavioral Sensitization and Conditioned Place Preference in Mice at JoVE.com

Assessment of Cocaine-induced Behavioral Sensitization and Conditioned Place Preference in Mice

This protocol is intended to enable researchers to conduct experiments designed to test these aspects of addiction using the conditioned place preference and locomotor behavioral sensitization assays.

코카인 유발 행동 과민성 및 마우스의 조건 장소 선호도 평가

It is thought that rewarding experiences with drugs create strong contextual associations and encourage repeated intake. In turn, repeated exposures to ...

Research

JoVE Journal

Behavior

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

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 ...

연구

행동분석학

Behavioral Assessment of the Aging Mouse Vestibular System

노화 마우스 전정 시스템의 행동 평가

Age related decline in balance performance is associated with deteriorating muscle strength, motor coordination and vestibular function. While a number of ...

A Procedure to Observe Context-induced Renewal of Pavlovian-conditioned Alcohol-seeking Behavior in Rats

절차 흰쥐의 파블로프 에어컨 알코올 추구 행동의 상황에 의한 갱신을 준수합니다

Environmental contexts in which drugs of abuse are consumed can trigger craving, a subjective Pavlovian-conditioned response that can facilitate ...

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

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

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 ...

Behavioral Assessment of Hearing in 2 to 4 Year-old Children: A Two-interval, Observer-based Procedure Using Conditioned Play-based Responses

4 년짜리 어린이 2에서 청각의 행동 평가 : 두 간격, 관찰자 ​​기반 절차 에어컨 플레이 기반의 응답을 사용하여

Collecting reliable behavioral data from toddlers and preschoolers is challenging. As a result, there are significant gaps in our understanding of human ...

Battery of Behavioral Tests Assessing General Locomotion, Muscular Strength, and Coordination in Mice

행동의 배터리 테스트 사정 일반 운동, 근육 강도, 그리고 조정 마우스

Behavioral testing is used in pre-clinical trials to assess the phenotypic effects and outcomes that a particular disease or treatment has on the ...

Reinstatement of Drug-seeking in Mice Using the Conditioned Place Preference Paradigm

바른된 장소 기본 패러다임을 사용 하 여 마우스에 마약 추구의 복직

The present protocol describes the Conditioned Place Preference (CPP) as a model of relapse in drug addiction. In this model, animals are first trained to ...

The Unpredictable Chronic Mild Stress Protocol for Inducing Anhedonia in Mice

마우스에 Anhedonia 유도 대 한 예측할 수 없는 만성 가벼운 스트레스 프로토콜

Depression is a highly prevalent and debilitating condition, only partially addressed by current pharmacotherapies. The lack of response to treatment by ...

A Conditioned Place Preference Protocol for Measuring Incubation of Craving in Rats

쥐에 갈망의 부 화를 측정 하기 위한 바른된 장소 설정 프로토콜

A major cause of repeated relapses is a craving for the drug. Drug craving increases progressively during the abstinence period, a phenomenon termed ...

A Behavioral Test Battery for the Repeated Assessment of Motor Skills, Mood, and Cognition in Mice

모터 기술, 분위기, 그리고 쥐에 있는 인식의 반복 된 평가 행동 테스트 배터리

Pharmacological and toxicological studies in neurodegeneration require comprehensive behavioral analysis in mice because motor dysfunctions and ...