Support provided by grant AA019462 to DAH and AA019884 to DDS.

Todos os procedimentos descritos aqui e no vídeo que o acompanha foram aprovados pelos Comitês de cuidado e uso de animais Institucional do Centro de Ciências da Saúde e do principal campus da Universidade do Novo México. 1. Pré-natal Ethanol Exposição <ol><li> Obter todos os materiais e produtos químicos necessários: Sacarina hidrato de sal de sódio, 190 etanol de prova (95% de álcool por volume), tubos de vidro de consumo de contas, borracha natural branco # 4 rolha de um buraco, um &quot;ponto de dobrar tubos de bola, os governantes de papel impresso a partir www .vendian.org / mncharity / pasta3 / paper_rulers /. </li><li> Obter ratos comprovadas adulto criador de um fornecedor ou fonte alternativa. Use criadores Long-Evans para os métodos e os dados representativos apresentados neste protocolo. Fêmeas pesam 125-150 g e 6-7 semanas de idade no momento da chegada de tal forma que eles são aproximadamente 9-10 semanas de idade na altura da reprodução (passo 1.5). Assegure-se que todas as barragens prospectivos são mães pela primeira vez. Certifique-se de que os homens são 12semanas de idade no momento da chegada e 15 semanas de idade no início do protocolo de reprodução. </li><li> Casa os animais individualmente em gaiolas de plástico a 22 ° C em um 12 horas de luz / escuridão cronograma reverso (luzes acesas 21:00-09:00 hr). Proporcionar o acesso a comida e água da torneira ad libitum durante todo o estudo, incluindo durante todas as sessões de consumo. Permitir que pelo menos uma semana para aclimatação à facilidade antes de prosseguir para a etapa 1.4. </li><li> Obter pesos corporal basal para cada rato fêmea. </li><li> Avalie os níveis de consumo pré-gravidez em ratas durante 2 semanas. Siga os passos 1.5.1-1.5.6 em ordem. <ol><li> Nos dias 1 e 2 encher um tubo de beber contendo solução de 0,066% de sacarina (sem etanol) em água da torneira. Para quantificar o consumo de apor uma régua de papel com mm de precisão para o tubo de beber antes de preenchê-lo. Utilizar este método, uma vez que reduz o erro de medição associado com tubos de pesagem para quantificar o consumo de etanol. Encha o tubo até a marca de 20 milímetros na régua. Comece quatro horas drinkisessões ng (passos 1.5.1-1.5.6 e 1,7) 1 h após o início da fase de obscuridade (10:00 horas) quando os níveis de actividade e de consumo são mais elevadas. </li><li> No fim de cada sessão de beber quantificar o volume da solução de sacarina que foi consumido. Para facilitar a medição, determinar o volume de solução por mm na régua de antecedência e converter mm para volumes. Para os tubos aqui recomendados cada mm corresponde a 0,366 ml. </li><li> Nos dias 3 e 4 encher um tubo com etanol a 2,5% (v / v) e solução de sacarina 0,066% em água da torneira. No fim de cada sessão de quantificar a quantidade de solução de etanol (peso por kg de peso corporal) que foi consumido. </li><li> No dia 5 e depois disso encher um tubo com 5% de etanol (v / v) e solução de sacarina 0,066% em água da torneira. No fim de cada sessão de quantificar a quantidade de solução de etanol (peso por kg de peso corporal) que foi consumido. </li><li> Após a conclusão da fase de pré-gravidez beber pesar os ratos e calcular o consum média etanolption e desvio padrão para todo o grupo. Remover os ratos para os quais o consumo médio é maior do que um desvio padrão acima ou abaixo da média do grupo do estudo para reduzir a variabilidade na ingestão voluntária durante a gravidez. </li><li> Atribuir os ratos restantes, quer o controle sacarina ou condições PAE tais que os níveis de consumo pré-gravidez são correspondidos, tanto quanto possível para os dois grupos. </li></ol></li><li> Dentro de 1-14 dias par cada rato fêmea com um macho reprodutor comprovado. Ratas não consomem etanol durante a fase de reprodução. Confirmar gravidez pela presença de um tampão vaginal, pesar o rato fêmea, e abrigar seu individualmente. Isto é definido como o dia gestacional 1. Opcionalmente, deixar as ratas com um reprodutor masculino para até 5 dias, após o que removem as fêmeas do estudo. Observação: Como o consumo de etanol durante a gravidez começa às 10:00 h na gestacional dia 1 de inspeção para a presença de um plug vaginal deve ser realizadaantes deste horário. </li><li> Fornecer soluções de etanol ou a sacarina durante 4 horas por dia (1000 a 1400 h) durante o período da gravidez. <ol><li> Começando no dia gestacional 1 proporcionar a rata quer com 0% ou 5% de etanol e solução de sacarina 0,066% na água da torneira com base no trabalho de grupo. Certifique-se de que o volume da solução de etanol desde 0% a ratos de controlo sacarina é comparado com a média do volume de solução etanólica a 5% de álcool consumido pelos ratos. No fim de cada sessão de quantificar a quantidade de solução de etanol (peso por kg de peso corporal) ou a sacarina que foi consumido. </li><li> Pesar barragens rato semanalmente para avaliar o ganho de peso materno. </li><li> Fornecer alimentos e água da torneira em todas as ocasiões, incluindo as sessões de beber. </li><li> Cessar os procedimentos de exposição ao etanol quando a prole nascem. Anote o número de filhotes vivos e do peso das crias ao nascer. Designar o dia do nascimento como pós-natal dia 0. </li><li> Abater o lixo para 10 filhotes ao redor dia pós-natal 2-3. Tentativa to manter uma proporção igual de macho para fêmeas em cada ninhada. Se isso não for possível, a abater 10 crias com um número desigual de machos e fêmeas. </li><li> Grave peso das crias de 2-3 dias após o nascimento. </li><li> Desmamar os animais no dia pós-natal aproximadamente 21-24 e casa em pares do mesmo sexo com um animal da mesma condição de tratamento pré-natal. Não use mais de 1-2 ratos de cada ninhada por experimento para limitar os efeitos potenciais de maca. </li></ol></li></ol> 2. Comportamento Social <ol><li> Obtenha e preparar todos os materiais e equipamentos necessários. <ol><li> Obter um aparelho para a interação social. Use uma câmara com uma parte superior aberta construída de material que é fácil de limpar e desinfetar. A parte da frente do aparelho deve ser coberto com plástico transparente rígido (~ 2 mm de espessura) para a filmagem. Linha as paredes interiores e chão com plástico transparente (~ 2 mm) para ajudar na limpeza e controle de odor. Coloque espelhos ao longo do wa interior de voltall para auxiliar a análise. NOTA: Para obter os dados representativos aqui relatado uma câmara de costume (95 cm de comprimento x 47 centímetros de largura × 43 cm de altura) com uma parte superior aberta e plástico rígido transparente (2 mm de espessura) que cobre o interior eo chão do aparelho foi usado. Espelhos foram colocados ao longo da parede traseira. As dimensões e materiais específicos não são críticos para a medição do comportamento social, no entanto, recomenda-se que as dimensões de ser suficientemente grande para garantir que a interacção social genuíno pode ser distinguido de outros comportamentos. Isto é, o aparelho deve ser suficientemente grande de modo a que os animais não são sempre em estreita proximidade um do outro. </li><li> Obter câmeras de vídeo capaz de gravar em condições de baixa ou nenhuma luz como todos filmagens é conduzido com pouca ou nenhuma iluminação ambiente dentro do espectro visível. Certifique-se de que a câmera tem uma resolução alta dentro do espectro infravermelho, no entanto, qualquer câmera capaz de gravar em ambientes escuros em sua nativamodo de noite ou com iluminadores infravermelhos adicionais deve ser suficiente. </li><li> Posição iluminadores infravermelhos em todo o aparelho para melhorar a iluminação do aparelho de gravação de vídeo. </li><li> Obter lascas de madeira de grau de laboratório (chip Aspen). </li><li> Obter uma escova, pá, dióxido de cloro e álcool isopropílico (70%) para a limpeza do aparelho e entre as sessões de controlo de odores. </li></ol></li><li> Aclimatação aparelhos e comportamento social <ol><li> Para esta fase da experiência, fazer as medições em ratos adultos que são pelo menos 90 dias de idade. </li><li> Antes de cada sessão remover todas as fichas madeiras, limpe o aparelho limpo com álcool isopropílico para controlar odores entre as sessões e fornecer lascas de madeira frescas. Certifique-se de que as aparas de madeira cobrir totalmente o fundo do aparelho. Limpar e higienizar o aparelho com um agente apropriado, tal como o dióxido de cloro, na medida do necessário. </li><li> Durante três dias consecutivos colocar um animal e sua gaiola-mate para ocâmara durante 30 minutos a aclimatar os animais ao aparelho. Durante as sessões de aclimatação todas as luzes da sala estão desligados. </li><li> No final da terceira sessão de casa aclimatação os animais individualmente em gaiolas novas com roupas de cama, comida e água durante 24 horas para motivar a interação social. </li><li> Interação social Record no dia seguinte 24 horas após os animais foram separados. <ol><li> Retire as aparas de madeira a partir do aparelho, limpar e higienizar com dióxido de cloro, limpar com álcool isopropílico para controlar odores, e substituir as aparas de madeira antes da sessão. </li><li> Coloque uma ou mais câmeras para registrar a interação. Posição, pelo menos, uma câmara em frente do aparelho de modo que os espelhos na parede traseira do aparelho pode proporcionar uma perspectiva adicional para análise. </li><li> Recuperar animais um de cada vez e mantenha o animal em frente da câmera para que as características originais do teste padrão da pele pode ser notado. Estes elementos de identificação dea pele pode ser utilizada para distinguir os ratos durante a análise, em vez de marcação artificialmente os animais. Nota: Como muitos aspectos do roedor interação social envolvem sinais olfativos e sentir o cheiro do parceiro, sempre que possível odores estranhos introduzem deve ser evitado. Ratos Long-Evans têm tipicamente alguma característica do padrão da pele que podem ser utilizados para distinguir um determinado par de animais. Para outros estirpes (por exemplo, ratos Sprague-Dawley) abordagens alternativas, tais como marcação da cauda com um corante sem perfume pode ser utilizado. É importante reconhecer que muitos comportamentos de interesse são direcionados para alvos específicos (por exemplo., Anogenital sniffing dirigido perto da base da cauda, ​​os ataques lúdicas dirigidas à nuca, ou ataques agressivos dirigidos para os flancos ou barriga). Marcando os animais mais próximo da ponta da cauda longe destes alvos de interesse é recomendada. </li><li> Grave vídeo da interação social por pelo menos 12 min.</li><li> Monitorar os animais para o combate ao longo da sessão. Se possível, observar os animais através de um monitor ou janela para que o pesquisador não está na sala durante a sessão. NOTA: A luta tem sido raramente observada em estudos com ratos adultos, no entanto, os animais devem ser monitorados durante toda a sessão. A sessão deve ser interrompido se houver luta excessiva ou há sinais de danos ou lesões a um animal. </li></ol></li></ol></li><li> Codificação e análise comportamental. <ol><li> Identificar os seguintes comportamentos de interesse de acordo com o trabalho anterior com PAE 1,18. Quantificar a duração, frequência e latência para a primeira ocorrência dos seguintes comportamentos; luta livre (incluindo fixando), boxe, rastejando (cruzamento) sobre / sob o parceiro, anogenital sniffing, outro sniffing do corpo do parceiro (corpo sniffing), allogrooming (aliciamento do parceiro), criação e sniffing / cavando as lascas de madeira . Exemplos de cada comportamento são ilustrard no componente de vídeo deste artigo. </li><li> Quantificar os comportamentos sociais de interesse do vídeo. Obter a frequência, a duração total e latência para a primeira instância para cada comportamento de interesse. NOTA: A obtenção dessas medidas pode ser alcançado manualmente, no entanto, é recomendado quantificação destas medidas, utilizando análises computadorizadas de vídeo digitalizado. Um script Matlab (www.mathworks.com) para a reprodução de vídeo e quantificação dos comportamentos é fornecido como um complemento a este artigo. </li><li> Após a codificação é concluída para todos os ratos os dados duração, frequência e latência resultantes são analisados ​​com um pacote estatístico. </li></ol></li></ol>

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D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Impact of Combined Prenatal Ethanol and Prenatal Stress Exposure on Anxiety and Hippocampal-Sensitive Learning in Adult. 37 (12), 2039-2047 (2013).</li><li>Champagne, F. A., Francis, D. D., Mar, A., Meaney, M. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Variations+in+maternal+care+in+the+rat+as+a+mediating+influence+for+the+effects+of+environment+on+development.">Variations in maternal care in the rat as a mediating influence for the effects of environment on development.</a> Physiol. Behav. 79 (3), 359-371 (2003).</li><li>Hamilton, D. A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Patterns+of+social-experience-related+c-fos+and+Arc+expression+in+the+frontal+cortices+of+rats+exposed+to+saccharin+or+moderate+levels+of+ethanol+during+prenatal+brain+development.">Patterns of social-experience-related c-fos and Arc expression in the frontal cortices of rats exposed to saccharin or moderate levels of ethanol during prenatal brain development.</a> Behav. Brain Res. 214 (1), 66-74 (2010).</li><li>Barnett, S. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> A study in behaviour: Principles of ethology and behavioural physiology displayed mainly in the rat. , (1963).</li><li>Pellis, S. M., Pellis, V. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Play-fighting+differs+from+serious+fighting+in+both+target+of+attack+and+tactics+of+fighting+in+the+laboratory+rat+Rattus-Norvegicus.">Play-fighting differs from serious fighting in both target of attack and tactics of fighting in the laboratory rat Rattus-Norvegicus.</a> Aggressive Behav. 13 (4), 227-242 (1987).</li><li>Meaney, M. J., Stewart, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+descriptive+study+of+social-development+in+the+rat+(Rattus-Norvegicus).">A descriptive study of social-development in the rat (Rattus-Norvegicus).</a> Anim. Behav. 29 (1), 34-45 (1981).</li><li>Pellis, S. M., Pellis, V. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+prejuvenile+onset+of+play+fighting+in+laboratory+rats+(Rattus+norvegicus).">The prejuvenile onset of play fighting in laboratory rats (Rattus norvegicus).</a> Dev. Psychobiol. 31 (3), 193-205 (1997).</li><li>Himmler, B. T., Pellis, V. C., Pellis, S. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Peering+into+the+dynamics+of+social+interactions:+measuring+play+fighting+in+rats.">Peering into the dynamics of social interactions: measuring play fighting in rats.</a> J. Vis. Exp. (71), e4288-e4288 (2013).</li><li>Panksepp, J., Siviy, S., Normansell, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+psychobiology+of+play+:+Theoretical+and+methodological+perspectives.">The psychobiology of play : Theoretical and methodological perspectives.</a> Neurosci. Biobehav. Rev. 8 (4), 465-492 (1984).</li><li>Siviy, S. M., Panksepp, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sensory+modulation+of+juvenile+play+in+rats.">Sensory modulation of juvenile play in rats.</a> Dev. Psychobiol. 20 (1), 39-55 (1987).</li><li>Pellis, S. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+effects+of+orbital+frontal+cortex+damage+on+the+modulation+of+defensive+responses+by+rats+in+playful+and+nonplayful+social+contexts.">The effects of orbital frontal cortex damage on the modulation of defensive responses by rats in playful and nonplayful social contexts.</a> Behav. Neurosci. 120 (1), 72-84 (2006).</li><li>Blanchard, R. J., Blanchard, D. C., Takahashi, T., Kelley, M. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Attack+and+defensive+behavior+in+albino-rat.">Attack and defensive behavior in albino-rat.</a> Anim. Behav. 25 (3), 622-634 (1977).</li></ol>

The authors have no conflicts of interest to disclose.

O paradigma de exposição pré-natal álcool descrito aqui envolve o consumo voluntário de etanol (5% v / v) por barragens de rato durante a gravidez. Há um número de protocolos de exposição dos animais não-humanos para etanol durante o desenvolvimento do cérebro representado na literatura, os quais diferem no que diz respeito ao tempo, dose, a duração e via de administração de etanol, bem como as espécies em estudo. Apesar de um tratamento exaustivo das vantagens de vários protocolos de exposição não ?...

Estima-se que 1-5% das crianças são diagnosticadas com Transtornos do Espectro Alcoólico Fetal (FASDs) 2, que incluem a Síndrome Alcoólica Fetal (SAF), FAS parcial (DFP), e ligados ao álcool neurodesenvolvimento transtornos (Arnds) 3. Os déficits de comportamento social e cognição estão entre os resultados adversos mais comuns observados em crianças com FASDs 4-7. Consequências negativas não se limitam a exposição pré-natal pesado de álcool (PAE), como moderado PAE que não leva ao déficit morfológicas, comportamentais e cognitivas conspícua característica do FAS pode causar relativamente sutil, mas mesmo assim persistente, déficit em humanos com FASDs 10/08 e animais não-humanos expostos ao etanol durante o desenvolvimento do cérebro 11. A importância de compreender as consequências neurobiológicas e comportamentais correspondentes de moderada PAE é sublinhada pelas estimativas atuais indicam que a grande maioria dos casos FASD estão dentro dos menos severe gama do espectro 12. Vários laboratórios independentes relataram alterações no comportamento social roedor relacionadas à exposição ao etanol durante o desenvolvimento do cérebro, incluindo a diminuição da investigação e interação 1,13-15, alteradas jogo 14,16,17, aumento das interações agressivas 17,18, alterações na capacidade de resposta a estímulos sociais 19-21 e déficits nas preferências alimentares socialmente adquiridos e memória de reconhecimento social 22. Déficits de comportamento social têm sido observadas após a exposição aos pesados ​​(concentrações de etanol no sangue (CEBs) ~ 300mg / dl) níveis mais moderados de etanol (CEBs ~ 80mg / dl) 1 ou 22,23, e através de uma ampla gama de parâmetros para outro significativo fatores, incluindo tempo de exposição, duração da exposição, e idade no momento da medição comportamental. Pesquisas anteriores já haviam demonstrado que as alterações nos aspectos específicos da int sociaiseraction na idade adulta discriminar ratos expostos a níveis moderados de álcool a partir de animais de controle expostos a sacarina 1,18. Em particular, moderada PAE tem sido consistentemente associada com aumentos robustos no wrestling, o que sugere aumentos de comportamento agressivo, e níveis mais baixos de investigação social (por exemplo, farejando do parceiro) na idade adulta. Por causa alterações no comportamento social são consequências confiáveis ​​de PAE, a quantificação do comportamento social seguinte PAE pode deter importância para progredir em direção a uma compreensão mais completa das bases neurais de alterações relacionadas à PAE no comportamento social e do desenvolvimento de abordagens de intervenção. O objetivo deste artigo e o vídeo associado é fornecer instruções sobre o protocolo PAE moderada e métodos para a quantificação do comportamento social em filhos adultos que têm seguramente distinguidos pré-natal da prole não expostos ratos expostos ao álcool.

<table border="0" cellpadding="0" cellspacing="0">
	<tbody>
		<tr>
			<td>Saccharin sodium salt hydrate</td>
			<td>Sigma</td>
			<td>S1002</td>
			<td></td>
		</tr>
		<tr>
			<td>190 proof ethanol</td>
			<td>Sigma</td>
			<td>493538</td>
			<td></td>
		</tr>
		<tr>
			<td>Beaded glass drinking tubes</td>
			<td>Fisher</td>
			<td>14-955K</td>
			<td></td>
		</tr>
		<tr>
			<td>Natural rubber white #4 stopper one hole</td>
			<td>Plasticoid</td>
			<td>LSG4M181</td>
			<td></td>
		</tr>
		<tr>
			<td>1&#34; bend tubes-ball point</td>
			<td>Ancare</td>
			<td>TD-199-3&#34;</td>
			<td></td>
		</tr>
		<tr>
			<td>Paper rulers</td>
			<td>N/A</td>
			<td>N/A</td>
			<td><a href="http://www.vendian.org/mncharity/dir3/paper_rulers">www.vendian.org/mncharity/dir3/paper_rulers</a></td>
		</tr>
		<tr>
			<td>Apparatus for social interaction</td>
			<td>Custom built</td>
			<td>N/A</td>
			<td>95 cm X 47 cm X 43 cm</td>
		</tr>
		<tr>
			<td>Video cameras</td>
			<td>N/A</td>
			<td>N/A</td>
			<td>Capable of recording low/no light conditions</td>
		</tr>
		<tr>
			<td>Infrared illuminators</td>
			<td>Vitek</td>
			<td>VT-IR1-12</td>
			<td></td>
		</tr>
		<tr>
			<td>Teklad laboratory grade sani-chips</td>
			<td>Harlan</td>
			<td>7090A</td>
			<td></td>
		</tr>
		<tr>
			<td>Brush and dustpan</td>
			<td>N/A</td>
			<td>N/A</td>
			<td></td>
		</tr>
		<tr>
			<td>Isopropyl alcohol</td>
			<td>Sigma</td>
			<td>W292907</td>
			<td></td>
		</tr>
		<tr>
			<td>Chlorine Dioxide (1.5 mg Tablets)</td>
			<td>Quiplabs</td>
			<td>N/A</td>
			<td>Prepare per manufacturer&#39;s recommendation</td>
		</tr>
	</tbody>
</table>

moderate prenatal alcohol exposure and quantification of social behavior in adult rats

Alterations in social behavior are among the major negative consequences observed in children with Fetal Alcohol Spectrum Disorders (FASDs). Several independent laboratories have demonstrated robust alterations in the social behavior of rodents exposed to alcohol during brain development across a wide range of exposure durations, timing, doses, and ages at the time of behavioral quantification. Prior work from this laboratory has identified reliable alterations in specific forms of social interaction following moderate prenatal alcohol exposure (PAE) in the rat that persist well into adulthood, including increased wrestling and decreased investigation. These behavioral alterations have been useful in identifying neural circuits altered by moderate PAE1, and may hold importance for progressing toward a more complete understanding of the neural bases of PAE-related alterations in social behavior. This paper describes procedures for performing moderate PAE in which rat dams voluntarily consume ethanol or saccharin (control) throughout gestation, and measurement of social behaviors in adult offspring.

Ao longo de muitos ciclos de reprodução ratas na condição etanol beber consistentemente uma média de cerca de 2,1 g / kg de etanol por sessão de beber 4 h. Rato barragens consumir cerca de metade do total de quatro horas durante os primeiros 15 a 30 minutos depois da introdução dos tubos de bebida, resultando numa concentração de etanol no soro materno de pico de cerca de 60 mg / dl, medida no ponto de tempo de 45 min. Durante o restante 3,5 h do período de ingestão, eles continuam a consumir etanol a 5%, a ...

Watch this Scientific Journal Video about Moderate Prenatal Alcohol Exposure and Quantification of Social Behavior in Adult Rats at JoVE.com

Moderate Prenatal Alcohol Exposure and Quantification of Social Behavior in Adult Rats

The goal of the protocol presented here is to describe procedures to expose rats to moderate levels of alcohol during prenatal brain development and to quantify resulting alterations in social behavior during adulthood.

Moderado pré-natal álcool Exposição e quantificação do comportamento social em ratos adultos

Alterations in social behavior are among the major negative consequences observed in children with Fetal Alcohol Spectrum Disorders (FASDs). Several ...

moderate-prenatal-alcohol-exposure-quantification-social-behavior

Research

JoVE Journal

Behavior

9.6K visualizações.  University of New Mexico. The goal of the protocol presented here is to describe procedures to expose rats to moderate levels of alcohol during prenatal brain development and to quantify resulting alterations in social behavior during adulthood. 

Vídeo: Moderate Prenatal Alcohol Exposure and Quantification of Social Behavior in Adult Rats

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 ...

Comportamento

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 ...

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.

Roedor Cérebro microinjeção para estudar substratos moleculares do comportamento motivado

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

The Social Dimension of Stress: Experimental Manipulations of Social Support and Social Identity in the Trier Social Stress Test

In many situations humans are influenced by the behavior of other people and their relationships with them. For example, in stressful situations supportive behavior of other people as well as positive social relationships can act as powerful resources to cope with stress. In order to study the interplay between these variables, this protocol describes two effective experimental manipulations of social relationships and supportive behavior in the laboratory. In the present article, these two manipulations are implemented in the Trier Social Stress Test (TSST)&#8212;a standard stress induction paradigm in which participants are subjected to a simulated job interview. More precisely, we propose (a) a manipulation of the relationship between different protagonists in the TSST by making a shared social identity salient and (b) a manipulation of the behavior of the TSST-selection committee, which acts either supportively or unsupportively. These two experimental manipulations are designed in a modular fashion and can be applied independently of each other but can also be combined. Moreover, these two manipulations can also be integrated into other stress protocols and into other standardized social interactions such as trust games, negotiation tasks, or other group tasks.

Previous research on the social dimension of stress has focused on two important variables: social identity and social support. This protocol introduces an effective experimental manipulation of these two social variables and describes their implementation in a standard stress induction paradigm (Trier Social Stress Test).

A Dimensão Social do Stress: Manipulações experimentais de apoio social e de identidade social no Trier Social Stress Teste

In many situations humans are influenced by the behavior of other people and their relationships with them. For example, in stressful situations ...

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 ...

Examining Recall Memory in Infancy and Early Childhood Using the Elicited Imitation Paradigm

The ability to recall the past allows us to report on details of previous experiences, from the everyday to the significant. Because recall memory is commonly assessed using verbal report paradigms in adults, studying the development of this ability in preverbal infants and children proved challenging. Over the past 30 years, researchers have developed a non-verbal means of assessing recall memory known as the elicited or deferred imitation paradigm. In one variant of the procedure, participants are presented with novel three-dimensional stimuli for a brief baseline period before a researcher demonstrates a series of actions that culminate in an end- or goal-state. The participant is allowed to imitate the demonstrated actions immediately, after a delay, or both. Recall performance is then compared to baseline or to performance on novel control sequences presented at the same session; memory can be assessed for the individual target actions and the order in which they were completed. This procedure is an accepted analogue to the verbal report techniques used with adults, and it has served to establish a solid foundation of the nature of recall memory in infancy and early childhood. In addition, the elicited or deferred imitation procedure has been modified and adapted to answer questions relevant to other aspects of cognitive functioning. The broad utility and application of imitation paradigms is discussed, along with limitations of the approach and directions for future research.

The elicited imitation procedure was established to examine the development of recall memory in infancy and early childhood. This procedure has been widely used to establish a solid foundation of the nature of recall memory in infancy and early childhood.

Examinando Chamada de Memória na infância e primeira infância Usando o provocou Imitação Paradigm

The ability to recall the past allows us to report on details of previous experiences, from the everyday to the significant. Because recall memory is ...

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.

O psicomotora Teste Rodent Vigilância (rPVT): um método para avaliar Neurobehavioral Desempenho em ratos e camundongos

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

Using Clicker Training and Social Observation to Teach Rats to Voluntarily Change Cages

Cage cleaning is a routinely performed husbandry procedure and is known to induce stress in laboratory rats. As stress can have a negative impact on well-being and can affect the comparability and reproducibility of research results, the amount of stress experienced by laboratory animals should be minimized and avoided when possible. Further, the direct contact between the rat and animal caretaker during the cage change bears hygiene risks and therefore possibly negatively impacts the well-being of the rats and the quality of the research. 
Our protocol aims to improve the routinely performed cage changing procedure. For this reason, we present a feasible protocol that enables rats to learn via clicker training and observation to voluntarily change to a clean cage. This training helps to reduce stress caused by the physical disturbance and handling associated with the cage changes and concurrently enables a reduction in direct contact between animal and animal caretaker after the training phase is completed.
The implementation of clicker training to rats is fast and easy. Rats are generally interested in the training and efficiently learn the desired behavior, which entails changing cages through a pipe. Even without training, the rats learn to perform the desired behavior by observation, as 80% of the observational learning group successfully changed cages when tested. The training further helps to establish a relationship of trust between trainer and animal. As hygiene and well-being are both very important in animal experiments, this protocol might also help to improve high-quality research.

This protocol introduces a method of cage change for rats via clicker training. Rats learn the desired behavior not only by direct training but also by observational learning. The implementation of this fast and easy protocol might help to improve well-being and hygiene in rodent facilities.

Usando o Clicker formação e observação Social para ensinar ratos para mudar voluntariamente gaiolas

Cage cleaning is a routinely performed husbandry procedure and is known to induce stress in laboratory rats. As stress can have a negative impact on ...

Integrating Computerized Linguistic and Social Network Analyses to Capture Addiction Recovery Capital in an Online Community

The article describes a new methodology designed with the aim of finding a comprehensive, unobtrusive, and accurate way of capturing social recovery capital development in online communities of recovery from alcohol and drug (AOD) addiction. Recovery capital was conceptualised as both engagement in the online recovery community and identification with the community. To measure recovery capital development, naturally occurring data were extracted from the social media page of a specific recovery program, with the page being set up as a resource for a face-to-face recovery program. To map engagement with the online community, social network analysis (SNA) capturing online social interaction was performed. Social interaction was measured through the linkages between the online contributors/members of the online community as represented by program clients, staff, and supporters from the broader community. To capture markers of social identification with the online community, computerised linguistic analysis of the textual data (content from posts and comments) was conducted. Recovery capital captured in this way was analysed against retention data (a proxy outcome indicator), as days spent in the (face-to-face) recovery program. The online data extracted was linked to participant data in regards to program retention to test prediction of a key recovery outcome. This approach allowed the examination of the role of online support communities and assessment of the association between recovery capital (developed via the online community of recovery) and recovery outcomes.

The article describes a novel approach for analyzing dynamic online social interactions (in an online context) exemplified by a study of an online community of recovery from alcohol and drug addiction.

Integrando análises informatizadas de redes linguísticas e sociais para capturar capital de recuperação de vício em uma comunidade online

The article describes a new methodology designed with the aim of finding a comprehensive, unobtrusive, and accurate way of capturing social recovery ...

A Complex Diving-For-Food Task to Investigate Social Organization and Interactions in Rats

For many species, where status is a vital motivator that can affect health, social hierarchies influence behavior. Social hierarchies that include dominant-submissive relationships are common in both animal and human societies. These relationships can be affected by interactions with others and with their environment, making them difficult to analyze in a controlled study. Rather than a simple dominance hierarchy, this formation has a complicated presentation that allows rats to avoid aggression. Status can be stagnant or mutable, and results in complex societal stratifications. Here we describe a complex diving-for-food task to investigate rodent social hierarchy and behavioral interactions. This animal model may allow us to assess the relationship between a wide range of mental illnesses and social organization, as well as to study the effectiveness of therapy on social dysfunction.

This protocol describes a method of examining social hierarchy in a rat model. Rats perform a complex diving-for-food task in which they form a distinct hierarchy according to their willingness to dive underwater and swim to obtain a food pellet. This method is used to understand decision making and social relationships among highly social animals in small groups.