笔者想感谢安妮·李和莫琳·伯恩在他们的帮助下建立扩展的条件性位置偏爱协议。

所有的程序都按照威尔康乃尔医学院实验动物护理和使用委员会规则进行。 1。立体定向交货病毒载体的制备和设置<ol><li>如果通过一个非无菌的表面接触到被污染的仪器，无菌拭子，或手戴无菌手套，丢弃或重新消毒的仪器，用热珠灭菌，丢弃棉签或改变到新的无菌手套。 </li><li>将鼠标笼散热块上的床上用品，以温热为手术后的恢复。 </li><li>电动剃须刀剃须头，乙醇和碘消毒头皮。 </li><li>擦拭的耳朵的酒吧和嘴保持stereotax用70％的乙醇。 </li><li>使用5微升汉密尔顿注射器，吸取5微升病毒载体（1×10 6病毒颗粒/微升），并设置注射器架stereotax的。 </li><li>微波炉加热垫，或打开一个电热垫第二名stereotax。用干净的吸水垫覆盖，以减少烫伤的危险。监视器小鼠体温（在97-99.5°F之间）频繁的整个过程。 </li><li>注入鼠标（重量在22-35克之间），氯胺酮（100毫克/千克）和甲苯噻嗪（20毫克/千克）的混合物。一旦鼠标被麻醉，掐尖尾检查鼠标的反射，和任何动作或回应，以确保鼠标是充分麻醉。 </li><li>剃须头的耳朵和眼睛之间。 </li><li>清洁光头交替乙醇和碘消毒棉签涂药。 </li><li>转移鼠标在stereotax加热垫。 </li></ol> 2。立体定向显微注射病毒载体<ol><li>整个手术，戴上无菌手套。如果在任何时候通过触摸表面非无菌手套被污染，更换手套。 </li><li>零牙杆和耳酒吧秤。 </li><li>设置齿牙杆。 </li><li>拧在枪口。如果摆动或晶须移动ment是注意到在任何时间，管理IP助推器剂量（约0.05毫升），氯胺酮（100毫克/千克）和甲苯噻嗪（20毫克/千克）的混合物。 </li><li>拧紧耳酒吧。 </li><li>润滑眼睛PuraLube。 </li><li>只是耳朵后面，用手术刀切开头皮。 </li><li>广场斗牛犬剪辑的角落头皮剥离皮肤远离颅骨。前囟门和lambda应该很容易看到。 </li><li>前囟门到拉姆达确保缝线直对齐。如果缝合头是歪的重新调整定位。 </li><li>检查确保所有设置上stereotax的归零。 </li><li>将前囟门在针尖上。 </li><li>测量所有的坐标为前囟和lambda。这包括背/腹内侧/外侧和前/后的坐标。 </li><li>调整打印头直至腹侧坐标为前囟和lambda内0.02毫米对方的。 </li><li>对齐后，前囟门走内侧的内侧/横向（M / L）的坐标。确保的左侧和右侧对齐相互0.02毫米的范围内。 </li><li>前囟门，并从那里再中心针尖移动到所需的前/后的坐标中的前/后方向（A / P）。 </li><li>如果有必要，在一个方向上的角针并确保被锁定stereotax。出去到左边M / L坐标（+ / - 1.52） </li><li>测量腹侧的位置作为参考使用后的井眼钻颅骨的顶部。 </li><li>钻一个井眼，用无菌的钻头上面的需要的坐标。检查针可以进入不间断的通过头骨。 </li><li>放下针到所需的腹侧坐标。 </li><li>一旦坐标达到下面浸0.015毫米，创建一个小口袋病毒载体〜10秒。 </li><li> 〜0.1微升/分钟的速率注入所需体积的病毒载体。等待3分钟的病毒载体完全扩散。提高针尖0.015毫米，等待2分钟。 </li><li>针（如果这样做有角度的注射）浸在相反的方向。再次，记录腹侧位置作为参考的头骨顶部。 </li><li>重复相同的步骤，在第一侧上的相对侧上（如果这样做双侧注射）注射病毒载体。 </li><li>两个孔骨蜡封住他们使用的无菌棉签头木端。 </li><li>缝合头皮。 </li><li>应用神经阻滞伤员。 </li><li>拔下鼠标从stereotax和地方回暖笼加热块，45分钟恢复。 </li><li>监测疼痛的迹象，包括：整体经济活动下降或烦躁不安，减少食物和水的消耗或提高发声。可以施用丁丙诺啡（0.05 - 0.2毫克/公斤），每天两次一个星期，如果有必要的疼痛。可接受的替代品，包括：哌替啶（20-60毫克/公斤），喷他佐辛（10毫克/千克），纳布（4-8毫克/公斤）。 </li></ol> 3。条件性位置偏爱：习得N，消光及修复<ol><li>将小鼠到中央室的三腔的位置偏爱设备（医学Associates公司，圣奥尔本斯，VT，USA）60秒习惯一段断头台门关闭。 </li><li>习惯之后，开铡门，让小鼠自由探索使用：第四MedPC软件集团（Med Associates公司）记录在每个室花费的时间为1200秒，所有3室。 </li><li>根据他们的表现在预测试空调室分配老鼠。有偏见的设计，对药品监督管理局在随后的3天，在基准测试前的车厢是最首选。 </li><li>空调在未来3天中，可卡因（10毫克/公斤; IP）注入老鼠在上午的会议，并限制他们在分配给他们的室1,200秒紧随注射。返回自己的家乡笼在会议结束后的小鼠。 4小时后，注入小鼠，用生理盐水（0.01毫升/克体重）和字串，它们为1200秒午后到对面的腔室中。 </li><li>要测试的收购，在中央室断头台门关闭到位小鼠60秒习惯。打开的断头台门和允许自由探索为1200秒，花费的时间在第四MedPC软件集团（Med Associates公司）的所有记录室。偏好计算减去可卡因成对室中花费的时间量从盐水成对的腔室中使用了大量的时间。 </li><li>要熄灭的地方偏好，让小鼠自由探索的两个20分钟的会议每天在同一设备，分离由至少2个小时，开始24小时后，收购测试。在所有腔室中使用了时间，应再次记录和应与每个消光会话初始CPP的幅度。 </li><li>确定小鼠是否已经熄灭了药物引起的偏好通过评估是否偏好相提并论满足特殊室连续跨越两个灭绝天明显低于收购偏好得分。 </li><li>将恢复原状小鼠用吸剂量的可卡因（5或10毫克/千克;的ip），并重新暴露的装置，允许自由探索，并记录在每个隔室中花费的时间。 </li><li>管理Euthasol的安乐死溶液（150毫克/千克）。 transcardially灌注小鼠用4％多聚甲醛（PFA），并进行组织学检查来验证正确的显微注射的位置。 </li></ol>

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什么都没有透露。

区域和时间特异性基因消融治疗通过立体显微注射病毒载体结合卜蜂，包括灭绝和恢复阶段允许进行调查的具体基因的作用，以三个不同的阶段类似上瘾的行为。虽然条件性敲除利用传统的Cre-LoxP系统实现提供了空间 - 时间限制基因消融，立体定向显微注射Cre重组小鼠骨髓小鼠成离散的脑区，允许更严格控制基因在何时何地被淘汰。此外，这种情况下，在开发过程中发生的代偿性改变，这有时会?...

可卡因是一种高度加强精神兴奋剂。反复接触后，一些发生在分子和细胞适应奖励相关的大脑电路，被认为是导致强迫性药物寻求行为，促使高的复发率，构成了严重的临床问题1。可卡因发挥这些长期持久的行为通过调节基因表达的影响。要研究长期使用可卡因的适应问题，潜伏期的啮齿动物模型已被广泛使用。其中一个这样的模型是条件性位置偏爱（CPP）的范式。这种模式涉及一个有学问的发展之间的关联以前中性的环境和可卡因的奖赏。可卡因与特定室的几个配对后，动物被允许自由地探索可卡因成对和非可卡因配对环境，如果他们愿意的药物配对室，他们说已经获得了可卡因诱发的放置ê偏好。此外，消退训练期间，这种范式可以用来研究可卡因寻求行为的特定上下文恢复。 相比，像上瘾的行为的其他行为模型，如精神运动性的敏化作用，这是用来研究长期可卡因诱导的行为和分子可塑性的2,3和自行给药（SA），被认为是更准确地模仿上瘾的类似的行为在人类中发现，卜蜂范式是一个简单的过程，研究可卡因情境学习4。 CPP协议，可以方便地扩展到包括灭绝和恢复阶段，类似于SA，允许调查机制，基本药物的渴求和复发5-7，这被认为是发生在人类的药物复述方面寻找行为和药物-线索诱发复发8-10。 一种机制是基础可卡因诱导的行为可塑性，是每个不同的阶段，CPP，包括收购，消光，复职的特点，是在不同的脑区激活基因表达的独特的签名。直接测试奖励相关的大脑区域内的基因介导可卡因诱发行为的变化，它是有用的，能够选择性地在特定区域的方式来操纵他们。实现这一目标的方法之一是microinject重组腺相关病毒（腺相关病毒）载体，表达Cre重组酶的小鼠骨髓小鼠LoxP位点两侧的靶基因的老鼠分为不同的脑区，采用立体定向手术。此方法允许非常精确的时间和区域的控制基因在何时何地被烧蚀，在两个分割和未分化的神经元，而不会引起免疫反应11-13。这种控制水平是一个重要的小鼠骨髓小鼠繁殖瓦特比传统的Cre-LoxP位技术的优势第i个小鼠表达Cre重组酶的内源基因的启动子的控制下，在这种基因缺失的定时和分布，可以更严格的调控。另外，病毒载体介导的基因敲除的情况下，潜在的发展，可能会发生使用传统的基因敲除策略的补偿效应。 除了可卡因CPP，这里描述的小鼠骨髓的小鼠的大脑注射Cre重组腺相关病毒特定基因可应用于无处不在的行为范式，涉及到不同的阶段，包括自我管理​​和精神致敏评估的作用。例如，我们的实验室已利用Cre重组腺相关病毒技术研究的作用钙1.2 L-型钙离子通道的可卡因精神运动性过敏14。具体来说，腺相关病毒注射到Cre重组酶基因编码钙1.2两侧装接loxP小鼠伏隔核（NAC），以证明钙v </suB&gt; 1.2在这个区域介导的表达阶段的精神运动致敏14,15。然而，腺相关病毒酶Cre的策略不能使用，如果不存在与感兴趣的基因的小鼠骨髓小鼠，评估L-型钙1.3的Ca 2 +通道的精神运动性致敏作用时，我们的经验。因此，限制使用腺相关病毒的Cre呈现本身，如果条件不存在一个特定基因的小鼠。然而，rAAVs，击倒靶基因表达的siRNA可以用来检查CA 1.3通道14,15的作用，因为我们已经做了。 微量注射腺相关病毒的Cre小鼠骨髓小鼠成离散的脑区，然后测试他们在CPP范式允许调查调解类似上瘾的行为的各个阶段，他们行事的特定基因。使用这种范式反复可卡因管理在本质上帮助我们了解如何劫持BRAINS奖励电路造成适应不良分子信号传导通路和基因表达的变化，导致上瘾的状态1,16,17。

<table border="1">
	<tbody>
		<tr>
			<td>Name of Reagent/Material</td>
			<td>Company</td>
			<td>Catalog Number</td>
			<td>Comments</td>
		</tr>
		<tr>
			<td>Conditioned place preference activity chambers</td>
			<td>Med Associates, Inc., St. Albans, VT, USA</td>
			<td>MED-CPP-MS</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Stereotaxic alignment system for mouse</td>
			<td>David Kopf Instruments, Tujunga, CA, USA</td>
			<td>model 900</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Hamilton syringes</td>
			<td>Hamilton Company, Reno, Nevada, USA</td>
			<td>7634-01</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>rAAV2-Cre-GFP</td>
			<td>Vector BioLabs, Philadelphia, PA, USA</td>
			<td>7016</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>rAAV2-GFP</td>
			<td>Vector BioLabs, Philadelphia, PA, USA</td>
			<td>7004</td>
			<td>&nbsp;</td>
		</tr>
	</tbody>
</table>

stereotaxic microinjection of viral vectors expressing cre recombinase to study the role of target genes in cocaine conditioned place preference

显微注射重组腺相关病毒（腺相关病毒）载体表达Cre重组酶分为不同的选择性敲除基因的小鼠的大脑区域允许增强时间和特定区域控制基因缺失，现有的方法相比。 ，虽然有条件删除也可以实现通过交配小鼠表达Cre重组酶的小鼠骨髓基因小鼠的特定基因的启动子的控制下，立体显微注射允许针对离散的大脑区域在实验者的利益确定的时间点。在可卡因条件性位置偏爱，以及其他可卡因的行为范式，如自行服用或精神运动性过敏，可以涉及撤回，的灭绝和/或恢复阶段的背景下，这种技术是特别有用的，在探索这些不同的靶基因的独特贡献可卡因诱发塑性行为模型阶段。具体而言，这种技术允许选择性消融治疗的靶基因的行为在离散阶段，以测试他们的贡献，跨越时间的行为。最终，这种认识可以更有针对性的治疗药物，最好能够解决自己每个阶段成瘾行为的最有力的风险因素。

卜蜂国际执行CPP对显微注射小鼠后，应验证，队列控制注入（RAAV-GFP）小鼠通常优先收购药物配对室（ 图1A，1B）。小鼠被认为已取得当可卡因偏好（时间花在可卡因配对室减去花费的时间在生理盐水配对室）是一个特定的腔偏好明显高于基线偏好得分（ 图1B，甲对乙 ） 。控制注入小鼠体内，作为一个队列，如果不显示可卡因配对室的偏好，或显示反?...

Watch this Scientific Journal Video about Stereotaxic Microinjection of Viral Vectors Expressing Cre Recombinase to Study the Role of Target Genes in Cocaine Conditioned Place Preference at JoVE.com

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

本文介绍如何microinject病毒载体到小鼠大脑中，然后在条件性位置偏爱的范例，包括收购，灭绝和恢复阶段测试。

立体定向显微注射病毒载体表达Cre重组酶研究的靶基因的作用在可卡因条件性位置偏爱

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

stereotaxic-microinjection-viral-vectors-expressing-cre-recombinase

Research

JoVE Journal

Behavior

18.5K 次观看.  Weill Cornell Graduate School of Biomedical Sciences. 本文介绍如何microinject病毒载体到小鼠大脑中，然后在条件性位置偏爱的范例，包括收购，灭绝和恢复阶段测试。

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

Stereotactic Injection of MicroRNA-expressing Lentiviruses to the Mouse Hippocampus CA1 Region and Assessment of the Behavioral Outcome

MicroRNAs (miRNAs) are small regulatory single-stranded RNA molecules around 22 nucleotides long that may each target numerous mRNA transcripts and dim an entire gene expression pathway by inducing destruction and/or inhibiting translation of these targets. Several miRNAs play key roles in maintaining neuronal structure and function and in higher-level brain functions, and methods are sought for manipulating their levels for exploring these functions. Here, we present a direct in vivo method for examining the cognitive consequences of enforced miRNAs excess in mice by stereotactic injection of miRNA-encoding virus particles. Specifically, the current protocol involves injection into the hippocampal CA1 region, which contributes to mammalian memory consolidation, learning, and stress responses, and offers a convenient injection site. The coordinates are measured according to the mouse bregma and virus perfusion is digitally controlled and kept very slow. After injection, the surgery wound is sealed and the animals recover. Lentiviruses encoding silencers of the corresponding mRNA targets serve to implicate the specific miRNA/target interaction responsible for the observed effect, with na&iuml;ve mice, mice injected with saline and mice injected with "empty" lentivirus vectors as controls. One month post-injection, the animals are examined in the Morris Water Maze (MWM) for assessing their navigation learning and memory abilities. The MWM is a round tank filled with colored water with a small platform submerged 1 cm below the water surface. Steady visual cues around the tank allow for spatial navigation (sound and the earth's magnetic field may also assist the animals in navigating). Video camera monitoring enables measuring the route of swim and the time to find and amount the platform. The mouse is first taught that mounting the hidden platform offers an escape from the enforced swimming; it is then tested for using this escape and finally, the platform is removed and probe tests examine if the mouse remembers its previous location. Repeated tests over several consecutive days highlight improved performance of tested mice at shorter latencies to find and mount the platform, and as more direct routes to reach the platform or its location. Failure to show such improvement represents impaired learning and memory and/or anxiety, which may then be tested specifically (e.g. in the elevated plus maze). This approach enables validation of specific miRNAs and target transcripts in the studied cognitive and/or stress-related processes.

MicroRNAs have significant roles in brain structure and function. Here we describe a method to enforce hippocampal miRNA over-expression using stereotactic injection of an engineered miRNA-expressing lentivirus. This approach can serve as a relatively rapid way to assess the in vivo effects of over-expressed miRNAs in specific brain regions.

立体定向注射的microRNA表达慢病毒的小鼠海马CA1区和行为结果的评估

MicroRNAs (miRNAs) are small regulatory single-stranded RNA molecules around 22 nucleotides long that may each target numerous mRNA transcripts and dim an ...

科研

行为学

A Procedure to Study the Effect of Prolonged Food Restriction on Heroin Seeking in Abstinent Rats

In human drug addicts, exposure to drug-associated cues or environments that were previously associated with drug taking can trigger relapse during abstinence. Moreover, various environmental challenges can exacerbate this effect, as well as increase ongoing drug intake.
The procedure we describe here highlights the impact of a common environmental challenge, food restriction, on drug craving that is expressed as an augmentation of drug seeking in abstinent rats.
Rats are implanted with chronic intravenous i.v. catheters, and then trained to press a lever for i.v. heroin over a period of 10-12 days. Following the heroin self-administration phase the rats are removed from the operant conditioning chambers and housed in the animal care facility for a period of at least 14 days. While one group is maintained under unrestricted access to food (sated group), a second group (FDR group) is exposed to a mild food restriction regimen that results in their body weights maintained at 90% of their nonrestricted body weight. On day 14 of food restriction the rats are transferred back to the drug-training environment, and a drug-seeking test is run under extinction conditions (i.e.&#160;lever presses do not result in heroin delivery).
The procedure presented here results in a highly robust augmentation of heroin seeking on test day in the food restricted rats. In addition, compared to the acute food deprivation manipulations we have used before, the current procedure is a more clinically relevant model for the impact of caloric restriction on drug seeking. Moreover, it might be closer to the human condition as the rats are not required to go through an extinction-training phase before the drug-seeking test, which is an integral component of the popular reinstatement procedure.

A procedure that allows a demonstration of robust augmentation of drug seeking in food-restricted rats is described. Following heroin self-administration training, rats go through an abstinence period, in a drug-free environment, during which they are mildly food restricted. Drug seeking is then tested in the drug-associated environment.

一个过程来研究延长食品的限制对海洛因寻求戒断大鼠的影响

In human drug addicts, exposure to drug-associated cues or environments that were previously associated with drug taking can trigger relapse during ...

Fat Preference: A Novel Model of Eating Behavior in Rats

Obesity is a growing problem in the United States of America, with more than a third of the population classified as obese. One factor contributing to this multifactorial disorder is the consumption of a high fat diet, a behavior that has been shown to increase both caloric intake and body fat content. However, the elements regulating preference for high fat food over other foods remain understudied.
To overcome this deficit, a model to quickly and easily test changes in the preference for dietary fat was developed. The Fat Preference model presents rats with a series of choices between foods with differing fat content. Like humans, rats have a natural bias toward consuming high fat food, making the rat model ideal for translational studies. Changes in preference can be ascribed to the effect of either genetic differences or pharmacological interventions. This model allows for the exploration of determinates of fat preference and screening pharmacotherapeutic agents that influence acquisition of obesity.

Dietary fat content influences both energy intake and body fat composition in mammals. By examining rats&#8217; preference for high fat food in a series of choice experiments, it is possible to test genetic differences and pharmacological interventions on their preference for high fat food.

脂肪偏好：饮食行为的一种新模型大鼠

Obesity is a growing problem in the United States of America, with more than a third of the population classified as obese. One factor contributing to ...

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

Environmental contexts in which drugs of abuse are consumed can trigger craving, a subjective Pavlovian-conditioned response that can facilitate drug-seeking behavior and prompt relapse in abstinent drug users. We have developed a procedure to study the behavioral and neural processes that mediate the impact of context on alcohol-seeking behavior in rats. Following acclimation to the taste and pharmacological effects of 15% ethanol in the home cage, male Long-Evans rats receive Pavlovian discrimination training (PDT) in conditioning chambers. In each daily (Mon-Fri) PDT session, 16 trials each of two different 10 sec auditory conditioned stimuli occur. During one stimulus, the CS+, 0.2 ml of 15% ethanol is delivered into a fluid port for oral consumption. The second stimulus, the CS-, is not paired with ethanol. Across sessions, entries into the fluid port during the CS+ increase, whereas entries during the CS- stabilize at a lower level, indicating that a predictive association between the CS+ and ethanol is acquired. During PDT each chamber is equipped with a specific configuration of visual, olfactory and tactile contextual stimuli. Following PDT, extinction training is conducted in the same chamber that is now equipped with a different configuration of contextual stimuli. The CS+ and CS- are presented as before, but ethanol is withheld, which causes a gradual decline in port entries during the CS+. At test, rats are placed back into the PDT context and presented with the CS+ and CS- as before, but without ethanol. This manipulation triggers a robust and selective increase in the number of port entries made during the alcohol predictive CS+, with no change in responding during the CS-. This effect, referred to as context-induced renewal, illustrates the powerful capacity of contexts associated with alcohol consumption to stimulate alcohol-seeking behavior in response to Pavlovian alcohol cues.

A procedure to study the capacity of an alcohol associated environmental context to trigger the renewal of alcohol-seeking behavior in rats is described.

一个过程来观察大鼠巴甫洛夫空调酒精寻求行为的背景下引发的重建

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

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.

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.

啮齿动物脑显微注射学习动机行为的分子基板

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

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.

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.

可卡因诱导行为敏和小鼠位置偏爱评估

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

Place and Response Learning in the Open-field Tower Maze

This protocol describes how the Open-field Tower Maze (OFTM) paradigm is used to study spatial learning in rodents. This maze is especially useful for examining how rats learn to use a place- or response-learning to successfully navigate in an open-field arena. Additionally, this protocol describes how the OFTM differs from other behavioral maze paradigms that are commonly used to study spatial learning in rodents. The OFTM described in this article was adapted from the one previously described by Cole, Clipperton, and Walt (2007). Specifically, the OFTM was created to test spatial learning in rodents without the experimenter having to consider how &#8220;stress&#8221; might play a role as a confounding variable. Experiments have shown that stress-alone can significantly affect cognitive function1. The representative results section contains data from an experiment that used the OFTM to examine the effects of estradiol treatment on place- and response-learning in adult female Sprague Dawley rats2. Future studies will be designed to examine the role of the hippocampus and striatum in place- and response-learning in the OFTM.

The Open-field Tower Maze (OFTM) is a maze created to study the behavioral and neural mechanisms of spatial learning (e.g., place- or response-learning) in rats. This maze is especially useful for experimenters who want to use a non-stressful maze paradigm to investigate spatial learning in their research.

在打开场塔迷宫的地方，反应学习

This protocol describes how the Open-field Tower Maze (OFTM) paradigm is used to study spatial learning in rodents. This maze is especially useful for ...

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

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 acquire a conditioned place preference in a drug-paired compartment, and after the post-conditioning test, they perform several sessions to extinguish the established preference. The CPP permits the evaluation of the conditioned rewarding effects of drugs related to environmental cues. Then, the extinguished CPP can be robustly reinstated by the non-contingent administration of a priming dose of the drug, and by exposure to stressful stimuli. Both methods will be explained here. When the animal reinitiates the behavioral response, a reinstatement of the conditioned reward is considered to have taken place.
The main advantages of this protocol are that it is non-invasive, inexpensive, and simple with good validity criteria. In addition, it allows the study of different environmental manipulations, such as stress or diet, which can modulate relapse into drug seeking behaviors. However, one limitation is that if the researcher aims to explore the motivation and primary reinforcing effects of the drug, it should be complemented with self-administration procedures, as they involve operant responses of animals.

This protocol describes the Conditioned Place Preference (CPP) as a model of relapse. This procedure permits the measurement of relapse in laboratory animals, considering the impact of drug-associated environmental cues as craving and relapse in abstaining addicts is currently the focus of drug-abuse treatment programs.

用条件位置偏好范式恢复小鼠药物寻毒作用

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 ...

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

A major cause of repeated relapses is a craving for the drug. Drug craving increases progressively during the abstinence period, a phenomenon termed incubation of drug craving. Here, we describe a morphine conditioned place preference (CPP) protocol for measuring the incubation of craving in rats. In this protocol, a CPP paradigm mainly employing somatosensory cues is used to establish a long-term reward memory of morphine. A three-chamber CPP box that differs in the texture of the chamber floor is constructed. First, the animals are tested for their baseline preference to the two side chambers for three consecutive days. Then, they are injected intraperitoneally with morphine/saline and put into their non-preferred/preferred chamber for 45 min. After 6 days of conditioning, their preference to the side chambers is tested for 15 min at different time points after the last conditioning session. With this paradigm, the reward memory of morphine could last for at least 18 days. To test whether the above-mentioned protocol can model increased craving, the number of entrances into the two side chambers are counted during the abstinence period. The results show that the entrances increased, suggesting that the CPP paradigm could mimic the incubation of craving. Future studies can employ this model to study neural mechanisms underlying long-term memory and incubation of craving.

Here, a morphine conditioned place preference (CPP) protocol is described to measure incubation of craving in rats.

一种用于大鼠渴望培养的条件场所偏好协议

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

Assessing the Coherence of Parents' Short Narratives Regarding their Child Using the Five-Minute Speech Sample Procedure

Valid and efficient measures for assessing the quality of parent-child relationships are needed to facilitate research and evidence-based practice with parents. This paper focuses on such a method, namely Five-Minute Speech Sample-Coherence (FMSS-Coherence). In this method, a parent is asked to speak for five uninterrupted minutes about her/his child and their relationship. The resulted narrative is coded for coherence, namely the extent to which the parent provides&#160;in the narrative&#160;a clear, consistent, multidimensional and well-supported portrayal of the child. FMSS-Coherence is based on attachment research that shows that the coherence of parents&#8217; narratives is indicative of the quality of the parent-child relationship and child adjustment. It overcomes the limitations of attachment narrative measures which are typically labor intensive. FMSS-Coherence is less nuanced than extant attachment narrative measures. Yet, studies of families from different cultural backgrounds, across different child ages and in the context of typically developing children as well as children with special needs suggest that coherence can be reliably evaluated using the FMSS procedure. Furthermore, parents&#8217; FMSS-Coherence is associated with parenting quality and children&#8217;s socio-emotional adjustment. Thus, it holds promise for researchers and practitioners who seek a relatively time- and cost-effective method for assessing the coherence of parents&#8217; narratives regarding their child.

This paper introduces a method for assessing the coherence of parents' short narratives regarding their child and their relationship using the five-minute speech sample procedure.

使用五分钟言语样本程序评估家长对孩子的简短叙述的一致性

Valid and efficient measures for assessing the quality of parent-child relationships are needed to facilitate research and evidence-based practice with ...