This work was funded by the University of Michigan Department Of Psychiatry (U032826; JDM), the National Institute on Drug Abuse (NIDA; K08 DA037912-01; JDM) and the Department of Defense (DoD) National Defense Science and Engineering Graduate (NDSEG) Fellowship (CJF).

Todos os procedimentos foram aprovados pelo Comitê da Universidade sobre o Uso e tratamento dos animais (UCUCA; Universidade de Michigan, Ann Arbor, MI). 1. Equipamentos e Software <ol><li> Obter câmaras operativas comerciais e dispositivos. Veja a Tabela de Materiais. </li><li> Realizar treinamento em ratos câmaras de condicionamento operante (largura 24,1 cm × 20,5 cm de profundidade x 29,2 cm de altura), situada em armários de atenuação de som com ventiladores para fornecer o ruído ambiente. </li><li> Equipar cada câmara, com uma revista pelete (receptáculo), localizado centralmente na 21.6 cm de largura da parede frontal da câmara de operante 3 cm acima do chão de grelha de aço inoxidável. Certifique-se de que a revista pellet contém um sensor infravermelho que é capaz de detectar as entradas de cabeça para dentro do dispositivo. </li><li> Ligue a revista pellet a um distribuidor da pelota, que oferece pelotas de alimentos com sabor de bananas (45 mg). </li><li> De uma forma contrabalançada, posição um retrátil leVer (CS) de aproximadamente 2,5 cm de cada lado do recipiente de ressuspensão (esquerda ou direita) 3 cm acima do chão de grelha de aço inoxidável. </li><li> Certifique-se de que a alavanca contém uma luz de sinalização LED dentro dele para fazer a alavanca estendida visualmente mais evidente. </li><li> Se usando uma alavanca retráctil, calibrar a alavanca, colocando um peso de calibração 10 g na parte superior da alavanca, e depois ajustando o parafuso de calibração (localizado na parte superior do dispositivo) com uma chave de porca sextavada até que o peso por si só, é capaz de deprimir a alavanca. </li><li> Equipar uma casa de luz vermelha no topo da câmara na parede oposta do recipiente revista. Nota: Uma casa de luz vermelha é utilizado para aumentar a perceptibilidade da alavanca iluminado. </li></ol> 2. Formação PCA <ol><li> -House par adulto ratos Sprague Dawley (250-300 g) no momento da chegada e deixá-los sem ser perturbado por pelo menos dois dias na colônia de habitação, que é mantido em um 12 hr ciclo claro / escuro. </li><li> Gentilmente lidar com EACH rato durante pelo menos 1 min por dia durante dois dias, a fim de se aclimatar os ratos ao contacto humano. Fornecer ratos acesso ad libitum a comida e água na colônia habitacional. </li><li> Conhecimento ratos com as pelotas de alimentos com sabor de bananas, colocando pelotas do alimento nas suas gaiolas (aproximadamente 15 pelotas por rato) por dois dias, começando três dias antes do treino PCA. Nota: Se os ratos são alojados em pares, que é útil para dividir os peletes uniformemente sobre os dois lados da gaiola de origem. </li><li> Programa pré-treinamento Nota: Toda a nossa programação é realizada utilizando MEDState Notation (para Med Associates, software), eo manual está disponível online (mednr.com/pdf/doc003_MedPCProgramr.pdf). Programas são escritos como documentos de texto sem formatação (por exemplo, ASCII ou texto DOS) usando MedState notação, salvo com a extensão adequada (ou seja, [nome do arquivo] .mpc), então traduzido usando o programa MED-PC, Trans IV. Todos estes passos são fornecidos em maior detalhe na acima referida prógramação manual. <ol><li> Criar um programa de pré-treinamento que oferece cinqüenta pelotas do alimento em um tempo variável (VT) de programação 30 seg 5 min após o início do programa (ou seja, um pellet comida é entregue em média a cada 30 segundos, mas a entrega real varia aleatoriamente entre 0- 60 seg). </li><li> Escreva o programa pré-treinamento de tal forma que a casa da luz vermelha está acesa, mas a alavanca permanece retraída para a duração do programa (cerca de 25 min). </li><li> Realizar pré-treinamento e toda a experimentação subsequente durante o ciclo de luz. </li><li> Um dia antes do treino PCA, coloque cada rato em uma câmara operante, feche e tranque a porta da câmara, e fechar a porta do armário de atenuação do som. </li><li> Inicie o programa de pré-treinamento o mais rapidamente possível após a colocação de cada rato, na sua câmara. </li><li> Remover cada rato da sua câmara tão rapidamente quanto possível após o programa termina. </li><li> Registe o número de pelotas consumidas e excluir os ratos que não consumir todo opelotas de novos testes. </li></ol></li><li> Criar um programa de formação de APC que proporciona 25 ensaios em aproximadamente 40 minutos, começando 1 min após o início do programa. <ol><li> Escrever o programa para que, durante cada ensaio a alavanca é estendido para a câmara e iluminada por 8 s. </li><li> Programa retração alavanca para ser imediatamente seguido pela entrega independente de resposta de um pellet de alimentos para a revista pellet. </li><li> Separe cada teste em uma programação sec VT 90 (ie, extensão da alavanca e subsequente entrega pellet alimentos ocorre em média a cada 90 segundos, mas a entrega real varia aleatoriamente entre 30-150 seg). </li><li> Coloque cada rato numa câmara de condicionamento, feche e tranque a porta da câmara, e fechar a porta do armário de atenuação do som. </li><li> Inicie o programa de formação APC tão rapidamente quanto possível após a colocação de cada rato, na sua câmara. </li><li> Remover cada rato da sua câmara tão rapidamente quanto possível após o programa termina. </li><li> Eratos xclude que não consomem todas as pelotas de mais testes. </li></ol></li></ol> 3. Processamento de Dados <ol><li> Transferir informações de saída para uma folha de cálculo (por exemplo, um documento de planilha). </li><li> Criar um perfil que extrai informações de identificação (número de registro assunto, data de início, hora de início e número de caixa), bem como informações de saída (número e latência de prensas de alavanca e entradas de revistas). Nota: Nas experiências, as entradas do compartimento não-CS também são extraídas pelo perfil. Definir entradas de revistas não-CS como entradas para a revista que ocorrem fora do período sec CS 8 e durante o (ou seja, 60-120 seg) período VT 90 segundos durante o qual a alavanca não é estendido. </li><li> Além disso processar os dados, nivelando as apertar a alavanca e revistas latências entrada em 8 segundos (ou seja, a duração da apresentação CS). Isto pode ser conseguido por meio de folha de cálculo da função IF: IF (X&gt; 8,8, X). </li><li> calcular a pontuação depara viés de resposta (ou seja, o número total de pressões da alavanca e entradas de revista para uma sessão; [alavanca prensas - entradas revista] / [pressões da alavanca + entradas de revista]), pontuação de latência (ou seja, a latência média para executar uma imprensa alavanca ou entrada de revista durante uma sessão; [latência entrada revista - alavanca de imprensa latência] / 8), ea diferença de probabilidade (ou seja, a proporção de pressões da alavanca ou entradas de revista; [alavanca de imprensa probabilidade - revista entrada probabilidade]) 5. Nota: Com estas fórmulas, todas as pontuações deve variar entre 1,0 e -1,0. </li><li> Calcule a pontuação do índice PCA para cada rato pela média de seu viés de resposta, pontuação de latência, e a diferença de probabilidade dentro de cada sessão individual. Nota: A pontuação do índice PCA deve variar entre 1,0 e -1,0. </li></ol>

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The authors have nothing to disclose.

formação PCA pode ser usado para determinar a variação individual na tendência de atribuir incentivos relevância aos estímulos condicionados, que tem sido argumentado ser um componente importante de vulnerabilidade vício. Por exemplo, ele já havia sido demonstrado que STs atribuir incentivos relevância para ambos os alimentos e medicamentos pistas 6, e ambos géneros e sugestões relacionadas com as drogas ativam regiões cerebrais semelhantes dentro de um circuito amígdalo-estriato-tálamo na <sup...

A transição do uso de drogas ao vício envolve interações complexas entre 1,2 aprendizagem pavloviano e instrumental. Durante consumo de drogas e drogas de procura de comportamentos, ações e resultados são aprendidas através de processos instrumentais; no entanto, as relações entre estímulos (por exemplo, sugestões relacionadas com a droga) e recompensas também são aprendidas através de processos de Pavlov. Pistas de Pavlov adquirir valor preditivo, mas eles também podem adquirir incentivo valor motivacional 3, em que eles se tornam atraente, desejável, e capaz de promover e manter o comportamento recompensa-seeking 4. Muitos procedimentos que investigam recompensa aprendizagem em animais, no entanto, não permitir a disentanglement de preditivo em relação a aprendizagem motivacional incentivo. Em procedimentos de auto-administração de drogas, por exemplo, ambas as contingências instrumentais e pavlovianas são normalmente empregadas, de tal forma que os ratos aprendem a executar uma ação (por exemplo, nariz-poking, pressionando-alavanca, etc), a fim de receber um resultado (isto é, infusão intravenosa de drogas). O resultado gratificante também está emparelhado com estímulos de Pavlov (por exemplo, a iluminação de um porto nariz puxão, apresentação luz de sinalização, o som da bomba de infusão, sensação interoceptiva de fornecimento de fluido para a corrente sanguínea, etc.). Nem sempre é claro nestes procedimentos se pistas estão apoiando ações dirigidas a objetivos simplesmente por causa de sua relação preditiva com a recompensa, ou se sinais adquiriram propriedades motivacionais de incentivo da sua própria. Para isolar experimentalmente o valor motivacional incentivo do valor preditivo de estímulos de Pavlov, nosso laboratório usa um procedimento pavloviano abordagem condicionado (PCA), a fim de identificar os ratos que atribuem incentivos relevância para recompensar-relacionados pistas. Durante uma sessão de treinamento, um estímulo condicionado (CS; por exemplo, uma alavanca) resposta de forma independente prevê a Delivery de um estímulo não condicionado (US, por exemplo, uma pastilha de alimento). Ao longo de várias sessões de treino, três fenótipos desenvolver: Sinal de rastreamento (resposta condicionada CS-dirigido [CR]), a meta-tracking (CR US-dirigido), e uma resposta intermediária (ambos CRs). Para meta-trackers (GTs), o CS é utilizado como um preditor de entrega de recompensa; No entanto, para sign-trackers (STs), o CS é atribuído com saliência incentivo, tornando-se atraente e desejável. Nesta revisão, descrevemos o equipamento, instalação e processamento de dados necessários para realizar um procedimento de PCA. Além disso, nós fornecemos resultados representativos de treinamento PCA, traçar considerações experimentais importantes, e discutir a utilidade putativa de procedimentos PCA em investigar a dependência e outros transtornos neuropsiquiátricos.

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			<td>Standard Modular Operant Test Chamber with Modified Top for Rat</td>
			<td>Med Associates, Inc.</td>
			<td>ENV-008CT</td>
			<td>Top has access slot for drug delivery leashes or other harness systems; comes with ENV-008CTC (Cover Insert) for access slot</td>
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			<td>Expanded PVC Sound Attenuating Cubicle</td>
			<td>Med Associates, Inc.</td>
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			<td></td>
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			<td>PCI Operating Package for up to Sixteen Chambers</td>
			<td>Med Associates, Inc.</td>
			<td>MED-SYST-16</td>
			<td>Based on the number of operant chambers, MED-SYST-8 (PCI Operating Package for up to Eight Chambers) is also available</td>
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			<td>SmartCtrl</td>
			<td>Med Associates, Inc.</td>
			<td>DIG-716P2</td>
			<td></td>
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			<td>Universal Cable, 25&#8242; (7.6 m)</td>
			<td>Med Associates, Inc.</td>
			<td>SG-210CB</td>
			<td>Based on individual needs, SG-210CB-50 (Universal Cable, 50&#8242; [15.2 m]) is also available</td>
		</tr>
		<tr>
			<td>18&#8243; (45.7 cm) 3-Pin Mini-Molex</td>
			<td>Med Associates, Inc.</td>
			<td>SG-216A</td>
			<td>Based on individual needs, SG-216A-2 (2&#8242; [61.0 cm] 3-Pin Mini-Molex) is also available</td>
		</tr>
		<tr>
			<td>Power Cable, 25&#8242; (7.6 m)&#160;</td>
			<td>Med Associates, Inc.</td>
			<td>SG-210CP-25</td>
			<td></td>
		</tr>
		<tr>
			<td>Stainless Steel Grid Floor for Rat</td>
			<td>Med Associates, Inc.</td>
			<td>ENV-005</td>
			<td>Based on preference, ENV-005A (Stainless Steel Grid Floor for Mouse) is also available&#160;</td>
		</tr>
		<tr>
			<td>Reusable Waste Pan</td>
			<td>Med Associates, Inc.</td>
			<td>ENV-007-P3</td>
			<td></td>
		</tr>
		<tr>
			<td>Filler Panel Package for Standard Modular Test Chamber</td>
			<td>Med Associates, Inc.</td>
			<td>ENV-008-FP</td>
			<td></td>
		</tr>
		<tr>
			<td>Modular Pellet Dispenser for Rat, 45mg</td>
			<td>Med Associates, Inc.</td>
			<td>ENV-203M-45</td>
			<td></td>
		</tr>
		<tr>
			<td>Pellet Receptacle, Trough Type</td>
			<td>Med Associates, Inc.</td>
			<td>ENV-200R2M</td>
			<td>Based on preference, ENV-200R1M (Pellet Receptable, Cup Type) is also available&#160;</td>
		</tr>
		<tr>
			<td>Retractable Lever</td>
			<td>Med Associates, Inc.</td>
			<td>ENV-112CM</td>
			<td>Calibrated to 25 g; needs to be adjusted to 10 g before Pavlovian conditioned approach training</td>
		</tr>
		<tr>
			<td>Retractable Lever Cue Light</td>
			<td>Med Associates, Inc.</td>
			<td>ENV-112CML</td>
			<td></td>
		</tr>
		<tr>
			<td>House Light for Rat</td>
			<td>Med Associates, Inc.</td>
			<td>ENV-215M</td>
			<td></td>
		</tr>
		<tr>
			<td>100 mA Replacement Bulbs, Pack of 10</td>
			<td>Med Associates, Inc.</td>
			<td>SG-800R</td>
			<td>Red-colored; for use with ENV-215M (House Light for Rat)</td>
		</tr>
		<tr>
			<td>Pellets, 45 mg, Primate Purified Diet, Banana Flavor</td>
			<td>Bio-Serv</td>
			<td>F0059</td>
			<td>50,000/box&#160;</td>
		</tr>
	</tbody>
</table>

pavlovian conditioned approach training in rats

Pistas que são contingentemente emparelhado com incondicionados, estímulos gratificantes pode adquirir-se propriedades de recompensa por meio de um processo conhecido como a atribuição de relevância de incentivo, ou a transformação de estímulos neutros em atraente, &quot;queria&quot; estímulos capazes de motivar comportamento. Abordagem condicionado de Pavlov (PCA) desenvolve-se após a apresentação independente de resposta de um estímulo condicionado (CS; por exemplo, uma alavanca) que prevê a administração de um estímulo não condicionado (US, por exemplo, uma pastilha de comida) e pode ser utilizado para medir incentivo saliência durante a formação, três padrões. respostas da condicionado (CRS) pode desenvolver:. comportamento de rastreamento de sinal (CS-dirigido CR), o comportamento de rastreamento de meta (US-dirigido CR), e uma resposta intermediária (ambos CRs) Registe-trackers atribuir incentivos relevância para recompensar-relacionada pistas e são mais vulneráveis ​​a reintegração induzida por sugestão de drogas que procuram, bem como outros comportamentos relacionados com a dependência, tornando PCA um poteprocedimento entially valiosa para estudar a vulnerabilidade vício. Aqui, descrevemos os materiais e métodos utilizados para provocar um comportamento PCA de ratos, bem como analisar e interpretar o comportamento PCA em experiências individuais.

Verificou-se que 5-7 sessões diárias de formação PCA é suficiente para ratos fenótipo por STS, GT, e respondedores intermédios (IRS), embora os ratos podem ser formados ainda com base nas necessidades de um laboratório particular ou experiência. Fenotipagem é baseado nos resultados de índice PCA, que são calculados pela média do viés de resposta, a pontuação de latência, ea diferença probabilidade de sessões individuais como anteriormente descrito no protocolo. Fenóti...

Watch this Scientific Journal Video about Pavlovian Conditioned Approach Training in Rats at JoVE.com

Pavlovian Conditioned Approach Training in Rats

Here, we present a protocol to elicit Pavlovian conditioned approach behavior in rats. This procedure can be used to measure individual differences in the tendency to approach and attribute incentive salience to reward-related cues and investigate addiction vulnerability.

Formação Abordagem Condicionada pavloviano em Ratos

Cues that are contingently paired with unconditioned, rewarding stimuli can acquire rewarding properties themselves through a process known as the ...

pavlovian-conditioned-approach-training-in-rats

Research

JoVE Journal

Behavior

10.9K visualizações.  University of Michigan. Here, we present a protocol to elicit Pavlovian conditioned approach behavior in rats. This procedure can be used to measure individual differences in the tendency to approach and attribute incentive salience to reward-related cues and investigate addiction vulnerability. 

Vídeo: Pavlovian Conditioned Approach Training in Rats

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.

Microinjeção estereotáxica de vetores virais Expressando Cre Recombinase para estudar o papel de genes alvo em cocaína condicionado Lugar de Preferência

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

Comportamento

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.

Um procedimento para estudar o efeito da restrição alimentar prolongada em heroína Buscando em ratos abstinentes

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

Training Synesthetic Letter-color Associations by Reading in Color

Synesthesia is a rare condition in which a stimulus from one modality automatically and consistently triggers unusual sensations in the same and/or other modalities. A relatively common and well-studied type is grapheme-color synesthesia, defined as the consistent experience of color when viewing, hearing and thinking about letters, words and numbers. We describe our method for investigating to what extent synesthetic associations between letters and colors can be learned by reading in color in nonsynesthetes. Reading in color is a special method for training associations in the sense that the associations are learned implicitly while the reader reads text as he or she normally would and it does not require explicit computer-directed training methods. In this protocol, participants are given specially prepared books to read in which four high-frequency letters are paired with four high-frequency colors. Participants receive unique sets of letter-color pairs based on their pre-existing preferences for colored letters. A modified Stroop task is administered before and after reading in order to test for learned letter-color associations and changes in brain activation. In addition to objective testing, a reading experience questionnaire is administered that is designed to probe for differences in subjective experience. A subset of questions may predict how well an individual learned the associations from reading in color. Importantly, we are not claiming that this method will cause each individual to develop grapheme-color synesthesia, only that it is possible for certain individuals to form letter-color associations by reading in color and these associations are similar in some aspects to those seen in developmental grapheme-color synesthetes. The method is quite flexible and can be used to investigate different aspects and outcomes of training synesthetic associations, including learning-induced changes in brain function and structure.

Reading in color is a new method for training letter-color associations that are typically found only in grapheme-color synesthetes. It involves an implicit form of training that has potential for long-term associative training methods&#160;because the training is a byproduct of reading and any text can be colored.

Treinando Associações Carta cores sinestésicas por Leitura em cores

Synesthesia is a rare condition in which a stimulus from one modality automatically and consistently triggers unusual sensations in the same and/or other ...

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.

Um procedimento para Observar Renovação induzida por Contexto de pavloviano condicionado Comportamento Álcool-seeking em Ratos

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

Training Rats to Voluntarily Dive Underwater: Investigations of the Mammalian Diving Response

Underwater submergence produces autonomic changes that are observed in virtually all diving animals. This reflexly-induced response consists of apnea, a parasympathetically-induced bradycardia and a sympathetically-induced alteration of vascular resistance that maintains blood flow to the heart, brain and exercising muscles. While many of the metabolic and cardiorespiratory aspects of the diving response have been studied in marine animals, investigations of the central integrative aspects of this brainstem reflex have been relatively lacking. Because the physiology and neuroanatomy of the rat are well characterized, the rat can be used to help ascertain the central pathways of the mammalian diving response. Detailed instructions are provided on how to train rats to swim and voluntarily dive underwater through a 5 m long Plexiglas maze. Considerations regarding tank design and procedure room requirements are also given. The behavioral training is conducted in such a way as to reduce the stressfulness that could otherwise be associated with forced underwater submergence, thus minimizing activation of central stress pathways. The training procedures are not technically difficult, but they can be time-consuming. Since behavioral training of animals can only provide a model to be used with other experimental techniques, examples of how voluntarily diving rats have been used in conjunction with other physiological and neuroanatomical research techniques, and how the basic training procedures may need to be modified to accommodate these techniques, are also provided. These experiments show that voluntarily diving rats exhibit the same cardiorespiratory changes typically seen in other diving animals. The ease with which rats can be trained to voluntarily dive underwater, and the already available data from rats collected in other neurophysiological studies, makes voluntarily diving rats a good behavioral model to be used in studies investigating the central aspects of the mammalian diving response.

Detailed instructions are provided on how to train rats to voluntarily dive underwater through a 5 m long Plexiglas maze. Because the brains of rats have been very well characterized, voluntarily diving rats may help elucidate the central pathways of the mammalian diving response.

Ratos treinamento para Voluntariamente Dive Underwater: Investigações da Resposta Mergulho dos Mamíferos

Underwater submergence produces autonomic changes that are observed in virtually all diving animals. This reflexly-induced response consists of apnea, a ...

Protocol for Studying Extinction of Conditioned Fear in Naturally Cycling Female Rats

Extinction of conditioned fear has been extensively studied in male rodents. Recently, there have been an increasing number of studies indicating that neural mechanisms for certain behavioral tasks and response behaviors are different in females and males. Using females in research studies can represent a challenge because of the variation of gonadal hormones during their estrous cycle. This protocol describes well-established procedures that are useful in investigating the role of estrogen in fear extinction memory consolidation in female rats. Phase of the estrous cycle and exogenous estrogen administration prior to extinction training can influence extinction recall 24 hr later. The vaginal swabbing technique for estrous phase identification described here aids the examination and manipulation of naturally cycling gonadal hormones. The use of this basic rodent model may further delineate the mechanisms by which estrogen can modulate fear extinction memory in females.

Gonadal hormones such as estrogen modulate memory formation in a number of experimental paradigms including fear extinction memory. This protocol describes a set of methods for investigating the influence of gonadal hormones specifically during extinction in naturally cycling females, including estrous cycle monitoring and exogenous estrogen administration.

Protocolo de Estudos de Extinção do medo condicionado em Naturally Ciclismo ratas

Extinction of conditioned fear has been extensively studied in male rodents. Recently, there have been an increasing number of studies indicating that ...

The Modified Hole Board - Measuring Behavior, Cognition and Social Interaction in Mice and Rats

This protocol describes the modified hole board (mHB), which combines features from a traditional hole board and open field and is designed to measure multiple dimensions of unconditioned behavior in small laboratory mammals (e.g., mice, rats, tree shrews and small primates). This paradigm is a valuable alternative for the use of a behavioral test battery, since a broad behavioral spectrum of an animal&#8217;s behavioral profile can be investigated in one single test.
The apparatus consists of a box, representing the &#8216;protected&#8217; area, separated from a group compartment. A board, on which small cylinders are staggered in three lines, is placed in the center of the box, representing the &#8216;unprotected&#8217; area of the set-up. The cognitive abilities of the animals can be measured by baiting some cylinders on the board and measuring the working and reference memory. Other unconditioned behavior, such as activity-related-, anxiety-related- and social behavior, can be observed using this paradigm. Behavioral flexibility and the ability to habituate to a novel environment can additionally be observed by subjecting the animals to multiple trials in the mHB, revealing insight into the animals&#8217; adaptive capacities.
Due to testing order effects in a behavioral test battery, na&#239;ve animals should be used for each individual experiment. By testing multiple behavioral dimensions in a single paradigm and thereby circumventing this issue, the number of experimental animals used is reduced. Furthermore, by avoiding social isolation during testing and without the need to food deprive the animals, the mHB represents a behavioral test system, inducing if any, very low amount of stress.

This protocol describes the modified hole board, which is a behavioral test set-up that comprises the characteristics of an open field and a traditional hole board. This set-up enables the differential analysis of unconditioned behavior of small laboratory mammals as well as the analysis of cognitive abilities.

The Hole Board Modificado - Medir Comportamento, Cognição e Interação Social em camundongos e ratos

This protocol describes the modified hole board (mHB), which combines features from a traditional hole board and open field and is designed to measure ...

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.

Avaliação da sensibilização comportamental induzida por cocaína e preferência por lugar condicionado em Ratos

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

A Method for Evaluating the Reinforcing Properties of Ethanol in Rats without Water Deprivation, Saccharin Fading or Extended Access Training

Operant oral self-administration methods are commonly used to study the reinforcing properties of ethanol in animals. However, the standard methods require saccharin/sucrose fading, water deprivation and/or extended training to initiate operant responding in rats. This paper describes a novel and efficient method to quickly initiate operant responding for ethanol that is convenient for experimenters and does not require water deprivation or saccharin/sucrose fading, thus eliminating the potential confound of using sweeteners in ethanol operant self-administration studies. With this method, Wistar rats typically acquire and maintain self-administration of a 20% ethanol solution in less than two weeks of training. Furthermore, blood ethanol concentrations and rewards are positively correlated for a 30 min self-administration session. Moreover, naltrexone, an FDA-approved medication for alcohol dependence that has been shown to suppress ethanol self-administration in rodents, dose-dependently decreases alcohol intake and motivation to consume alcohol for rats self-administering 20% ethanol, thus validating the use of this new method to study the reinforcing properties of alcohol in rats.

This protocol describes a novel and efficient method to quickly initiate operant responding for ethanol in rats that, contrary to standard methods, does not require water deprivation or saccharin/sucrose fading to initiate responding.

Um método para avaliar as propriedades de reforço de etanol em ratos sem água Privação, Sacarina desvanecimento ou treinamento de Acesso Ampliado

Operant oral self-administration methods are commonly used to study the reinforcing properties of ethanol in animals. However, the standard methods ...

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.

Reintegração de busca de drogas em ratos utilizando o paradigma de preferência lugar condicionado

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 ...