We would like to dedicate this work to the memory of late Dr. William T. Greenough, a great mentor, a colleague and a friend. This work was supported by NIH/NIAAA grant number AA009838 and NIH/NIGMS COBRE: The Delaware Center for Neuroscience research grant 1P20GM103653 to AYK. We are grateful to the former and current members of Klintsova lab.

Declaração de Ética: O protocolo seguinte foi aprovado pelo Comitê Institucional de Animal Care and Use (IACUC) da Universidade de Delaware. 1. Exposição do Desenvolvimento (ou Modelo de Binge-como a exposição ao etanol) <ol><li> Em PD3, determinar o sexo de cada animal e cross-promover quaisquer animais se necessário para manter o tamanho da leitegada (8 animais) e distribuição por sexo (4 machos: 4 fêmeas) consistente dentro de cada ninhada. NOTA: É importante para manter o tamanho da leitegada e distribuição por sexo tão consistente quanto possível, para evitar problemas metodológicos. Embora este protocolo usa 8 filhotes (4 homens e 4 mulheres) por ninhada, os tamanhos da maca alternativos ou distribuições de sexo pode ser adaptado às necessidades do projeto experimental. </li><li> Por via subcutânea injetar uma pequena quantidade de tinta preta Índia nas patas para identificar os animais dentro de cada ninhada. </li><li> Pseudo-aleatoriamente atribuir ninhadas como experimentais (contendo 50% de controlo da entubado simulada (AE exposta-álcool) e 50% (SI) pups) ou mamar controle (SC) (animais que não sejam objecto de nenhuma intubação, recorte cauda, ​​ou protocolos de separação de PD 4-9, exceto para a pesagem diária e orelha-perfuração). <ol><li> Para manter o tamanho do grupo consistente, atribuir o dobro de ninhadas experimentais como ninhadas SC. </li></ol></li><li> Pesar cada animal, em seguida, devolvê-lo à sua gaiola. Pesagem de animais deve ocorrer diariamente durante o período de intubação (DP 4-9). <ol><li> Remova todo o lixo da barragem. </li><li> Coloque filhotes na almofada aquecida. </li><li> Grave o peso de cada filhote individual. </li></ol></li><li> Em PD4, depois de pesar cada animal calcular a quantidade de álcool necessária para um total de 5,25 g / kg / dia por cada animal (com base no peso das crias a partir do passo 1.4) 8. <ol><li> Administrar álcool como 11,9% de etanol-em-substituto do leite (vol / vol). </li></ol></li><li> A partir de 09:00, remover um filhotes de maca da mãe de cada vez. </li><li> Administrar o etanol-in-leite para cada filhote AE <s-se class = &quot;xref&quot;&gt; 34. </li><li> Sham-entubar cada filhote SI 8. </li><li> Repita os passos 1.5. através de 1.8. para cada ninhada experimental. </li><li> Duas horas após a primeira dose, repetir o procedimento de dosagem (1,5 passos através de 1,8) para uma segunda dose de álcool. </li><li> Uma hora e meia após a segunda dose de álcool (o ponto em que o conteúdo de álcool no sangue diária de pico é alcançada), recolher e sangue centrífuga dos filhotes AE e SI através da cauda de recorte para o futuro de álcool no sangue de análise de conteúdo 35. <ol><li> Recolha de 60 mL de sangue. </li><li> Coloque sangue num tubo de microcentrífuga de 1 mL. sangue centrífuga de 1,5 g durante 25 min. </li><li> Recolher cuidadosamente o soro sobrenadante do tubo de centrífuga e poupar para a análise de conteúdo de álcool no sangue futuro. </li></ol></li><li> Repita o procedimento de dosagem (passos 1.5 através de 1.8) usando leite em vez de etanol no leite para evitar déficits nutricionais de incapacidade de enfermagem em AEfilhotes. <ol><li> Execute um total de 2 doses de leite suplementares 2 h separados no PD 4. </li></ol></li><li> Repita os passos 1.4 a 1.12 (excepto para o passo 1.11) em DP 5-9. </li><li> Após a dose de leite suplementar final sobre PD9, orelha perfurar todos os filhotes para a identificação na gaiola CE. <ol><li> Coordenar ouvido perfurado com alguma medida de número de maca ou identificador (por exemplo, ninhadas ímpares dentro de uma coorte seria obter a sua orelha esquerda perfurada enquanto os animais de ninhadas de número par iria ficar orelha direita perfurada). Isto tornará mais fácil para identificar os animais na gaiola CE deverá vários animais de ninhadas diferentes têm o mesmo padrão pawmark. </li></ol></li></ol> 2. O desmame <ol><li> Em PD 23, casa todos os animais em gaiolas de 2-3. <ol><li> Assegurar que todos os animais alojados na mesma gaiola são do mesmo sexo. </li><li> Incluir um SC, um SI, e um animal por gaiola AE quando possível. </li><li> Minimize o número de companheiros de gaiola thAT são da mesma ninhada. </li><li> Certifique-se de todos os animais são capazes de acesso a alimentos e água. </li></ol></li></ol> Corrida 3. Roda <ol><li> No PD30, destinar metade das gaiolas com animais a WR. Casa estes animais em gaiolas com livre acesso à roda de corrida de aço inoxidável anexado. <ol><li> Certifique-se de que as rodas têm um contador para avaliar o número total de rotações. </li></ol></li><li> Pesar todos os animais em PD 30 e PD 36. </li><li> Verifique o número de voltas de cada roda às 9 da manhã todos os dias. </li><li> Deixar os animais em sua respectiva condição de habitação para 12 dias. </li></ol> 4. Complexidade Ambiental <ol><li> Prepare a gaiola CE, antes de 09:00 no dia que corresponde ao PD 42 para animais experimentais. <ol><li> Obter um 30 &quot;x 18&quot; x 36 &quot;gaiola de aço galvanizado. NOTA: A gaiola deve ter vários níveis, ser capaz de suportar o peso de vários ratos, ser preenchido com padrãoroupa de cama, e tem vários locais para prender garrafas de água e dispensadores de alimentos. </li><li> Coloque novela, objetos coloridos de tamanhos variáveis ​​e formas na gaiola. <ol><li> Coloque 6 grandes brinquedos na gaiola CE. Assegurar que cada brinquedo é grande o suficiente para 3 ou mais ratos para interagir com simultaneamente. </li><li> Coloque 6 brinquedos médias na gaiola CE. Assegurar que cada brinquedo é grande o suficiente para 3 - 4 ratos para interagir com simultaneamente. </li><li> Coloque um lote (pelo menos 20) de brinquedos mais pequenos na gaiola CE. </li><li> Use brinquedos de cores diferentes, formas, tamanho, etc. novidade é crítico para esta intervenção (ver discussão). </li></ol></li><li> Coloque dois pratos de comida em extremidades opostas da gaiola. </li><li> Colocar duas garrafas de água em extremidades opostas da gaiola. </li></ol></li><li> Às 9 horas da manhã de PD 42, pesar todos os animais e realocar os animais WR para a gaiola CE. Cada gaiola CE deve conter 9 - 12 animais. <ol><li> Certifique-se de que nenhum animal têm ambos o mesmo pawmark e orelha-punpadrões ch. </li></ol></li><li> Verifique todos os alimentos e água diariamente. </li><li> De dois em dois dias, remover os brinquedos da gaiola CE e substituí-los (de acordo com o passo 4.1.2.). </li><li> A cada três dias, limpar a gaiola CE. <ol><li> Retire os animais da jaula CE e colocá-los em gaiolas de detenção temporária de 2 - 3 animais. </li><li> Remover tudo do leito a partir do fundo da gaiola. </li><li> Retornam os mesmos brinquedos para a gaiola, a menos que este dia coincide com o cronograma de substituição brinquedo (de acordo com o passo 4.4.). </li><li> Substitua toda a comida e água. </li><li> Substitua os ratos na gaiola CE. </li></ol></li></ol> 5. Tissue Collect NOTA: recolha de tecidos (por exemplo, a perfusão com paraformaldeído) e de armazenamento (por exemplo, congelamento, embebidos em parafina) pode ser realizada com uma variedade de métodos. A seguir irá explicar o processo de perfusão com 4% de paraformaldeído em 0,1 M de solução salina tamponada com fosfato (4% de paraformaldeído em PBS) Solução a 8. Atenção: O paraformaldeído é cancerígeno e pode também causar irritação da pele, reacção alérgica cutânea ou lesões oculares. Use proteção para os olhos / pele adequado. <ol><li> Expor um rato de cada vez ao isoflurano para anestesiar levemente o animal. </li><li> Injectar por via intraperitoneal a ratos com 2 ml / kg de mistura de cetamina / xilazina (1,5 mL xilazina misturado com 10 mL de cetamina). NOTA: A cetamina e xilazina são ambos em concentrações de estoque de 100 mg / mL antes de combinar para mistura de injecção. </li><li> Depois de rato não é mais sensível, perfundir o animal com fosfato 0,1 M de solução salina tamponada (PBS; pH = 7,2) seguido por paraformaldeído a 4% em PBS (pH = 7,2). </li><li> Remover cérebro e armazenar em paraformaldeído a 4% em PBS a 4 ° C durante 48 h. </li><li> Após 2 dias, a transferência de solução de sacarose a 30% adicionado a 4% de paraformaldeído em PBS a 4 ° C. </li></ol>

The authors have nothing to disclose.

No protocolo anterior, nós demonstramos uma intervenção oportuno resgatar déficits neuroanatómicos após exposição neonatal álcool. Esta intervenção poderá ser usado como um agente terapêutico em outros modelos animais, devido à robustez de cada um dos componentes da intervenção. Actividade cardiovascular voluntária sob a forma de WR foi mostrado para beneficiar várias resultados comportamentais 38, 39 e induzir alterações plásticas funcionai...

Elevando em diferentes ambientes tem sido conhecido por causar mudanças em várias medidas de bem-estar neurológica. Muitos estudos olhar para os efeitos benéficos da criação em um ambiente complexo (CE) começando com uma pesquisa inovadora pela Diamond e Rosenzweig (por exemplo, 1, 2) e Greenough (Por exemplo, 3, 4). CE foi demonstrada para ter um efeito positivo sobre inegáveis alterações sinápticas e celulares no cérebro 5, 6, 7. CE pode afectar uma multiplicidade de regiões do cérebro, incluindo o hipocampo 8, 9 e córtex visual 10, 11, estriado ventral 12, 13, bemcomo função neuroimune ampla-cérebro (revisto em 14). Interesse particular tem desenvolvido a partir dos estudos sobre hipocampo quando foi demonstrado que a CE, pode aumentar a taxa de sobrevivência de células de grânulos de nascidos de adultos do giro denteado através de plasticidade dendrítica 9, 13. Este último ponto reuniu muito interesse devido ao crescente corpo de literatura indicando que o exercício cardiovascular promove neurogênese adulta, tanto no cérebro saudável e danificou 15, 16, 17, 18. Execução de rodas (W) é um fácil de implementar forma de atividade cardiovascular voluntário que tem sido mostrado para ser benéfico em modelos de roedores de distúrbios neurológicos ou envelhecimento 17, 19, 20. WR afecta a expressão de factores de crescimento<html> tanto no sistema nervoso central e periférico 21, 22, 23. Combinando (subsequentemente) WR e CE numa &quot;superintervention&quot; (WR-CE) (isto é, 12 dias de WR seguido por 30 dias em CE) proporciona um aumento robusto em adultos neurogenesis hipocampo e aumento da sobrevivência das células recém-proliferadas 8, o efeito que, no modelo animal de SAF não é alcançada pelos componentes individuais (ver abaixo). Desde ambos os componentes do WR-CE afectam uma diversidade de estruturas dentro do cérebro 13 (WR revisto em 22, CE revisto em 24), a execução dessa intervenção pode ser facilmente aplicada a modelos de roedores de ambos os modelos início da vida de desenvolvimento e posteriores do neurológica impairment (por exemplo, álcool exposição neonatal, envelhecimento, estresse precoce). nt &quot;&gt; Integração da WR-CE nos períodos adolescente e início de adultos (ou seja, dias pós-natal 30 - 72) pode melhorar alguns dos efeitos negativos de um modelo de rato de perturbações do espectro do alcoolismo fetal (FASDs) 8 Uma coleção de estudos. demonstraram que os roedores expostos ao álcool a partir do dia pós-natal (PD) de 4 a 9 de exibição déficits significativos em medidas neuroanatómicos como complexidade dendrítica 25, o desenvolvimento do cerebelo 26, 27 e capacidade de resposta neuroimmune 28, bem como manifestações de prejuízo no aprendizado e na memória 29, 30, 31 . Mesmo uma quantidade reduzida de exposição ao álcool dentro desta janela de tempo (ou seja, PD 7 a 9) pode levar a déficits de aprendizagem e memória em ratos adolescentes e adultos 32 enquanto algumas estruturas já não vê sigimpairment neuroanatômica signi- 27. Muitos desses déficits -, além de deficiências comportamentais em tarefas dependentes do hipocampo - foram atenuados após a exposição a este paradigma WR-CE 8, 33 ou WR sozinho 25, 31. Embora WR sozinho tem sido amplamente utilizada uma intervenção, a combinação de WR-CE ainda não foi utilizado na literatura, apesar da sua capacidade de manter os benefícios relativamente mais curto prazo de WR 8. Este artigo irá discutir a implementação da intervenção WR-CE durante a adolescência. Embora este paradigma é usado no contexto da exposição precoce álcool pós-natal, que pode ser introduzido para vários modelos de roedores para avaliar o potencial para o cérebro neuroplasticidade nos modelos de perturbações cerebrais.

<table border="0" cellpadding="0" cellspacing="0" width="698">
	<colgroup>
		<col />
		<col />
		<col />
		<col />
	</colgroup>
	<tbody>
		<tr height="42">
			<td height="42" style="height:42px;width:247px;">Female Time-pregnant Long Evans Rats</td>
			<td style="width:124px;">Envigo (Formerly: Harlan, Inc.)</td>
			<td style="width:136px;"></td>
			<td style="width:191px;">Average litter size is 8-10 pups</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:247px;">Black India Ink</td>
			<td style="width:124px;">Higgins (Chartpak, Inc.)</td>
			<td style="width:136px;">44201</td>
			<td style="width:191px;"></td>
		</tr>
		<tr height="84">
			<td height="84" style="height:84px;width:247px;">Syringes and Injection Needles</td>
			<td style="width:124px;">Becton, Dickinson and Company (BD)</td>
			<td style="width:136px;">Assorted</td>
			<td style="width:191px;">For injection of pawmarking ink, administration of milk-alcohol solution</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:247px;">Ear Punch</td>
			<td style="width:124px;">Kent Scientific Corporation</td>
			<td style="width:136px;">INS750076</td>
			<td style="width:191px;"></td>
		</tr>
		<tr height="63">
			<td height="63" style="height:63px;width:247px;">Running Wheels</td>
			<td style="width:124px;">Wahmann Labs</td>
			<td style="width:136px;"></td>
			<td style="width:191px;">Wahmann Running Wheel is discontinued. Substitute with&#160; One per cage</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:247px;">EC Cage</td>
			<td style="width:124px;">Martin&#39;s Cages, Inc.</td>
			<td style="width:136px;">R-695</td>
			<td style="width:191px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:247px;">Small EC Toys</td>
			<td style="width:124px;">Assorted</td>
			<td style="width:136px;"></td>
			<td style="width:191px;"></td>
		</tr>
		<tr height="63">
			<td height="63" style="height:63px;width:247px;">Medium EC Toys</td>
			<td style="width:124px;">Assorted</td>
			<td style="width:136px;"></td>
			<td style="width:191px;">Should be able to fit 1-2 rats inside of/ on top of object</td>
		</tr>
		<tr height="63">
			<td height="63" style="height:63px;width:247px;">Large EC Toys</td>
			<td style="width:124px;">Assorted</td>
			<td style="width:136px;"></td>
			<td style="width:191px;">Should be able to fit 3 or more rats inside of/on top of object</td>
		</tr>
	</tbody>
</table>

wheel running and environmental complexity as a therapeutic intervention in an animal model of fasd

O exercício aeróbico (por exemplo, roda de corrida (WR) amplamente utilizado em pesquisas com animais) impacta positivamente muitas medidas de potencial neuroplastic no cérebro, tais como taxas de neurogênese adulta, angiogênese e expressão de factores neurotróficos em roedores. Esta intervenção também foi mostrado para mitigar os aspectos comportamentais e neuroanatômicos de os impactos negativos da teratogens (ou seja, a exposição de desenvolvimento a álcool) e neurodegeneração relacionada com a idade em roedores. complexidade Ambiental (CE) foi mostrado para produzir inúmeros benefícios neuroplásticas em estruturas corticais e sub-corticais e pode ser acoplado com a roda de corrida para aumentar a proliferação e sobrevivência de novas células do hipocampo de adulto. A combinação destas duas intervenções fornece um &quot;superintervention&quot; robusto (WR-CE), que pode ser implementada numa variedade de modelos de roedores de distúrbios neurológicos. Vamos discutir a implementação de WR / CE e seu constituinteintervenções para utilização como uma intervenção terapêutica mais forte em ratos utilizando o modelo animal de exposição pré-natal ao álcool em humanos. Também vamos discutir quais elementos dos procedimentos são absolutamente necessários para as intervenções e quais podem ser alterados dependendo da pergunta ou instalações do experimentador.

A fim de avaliar o efeito da intervenção super, temos de olhar para os efeitos de cada um dos seus elementos constitutivos - WR e CE - em nossas medidas de interesse. As Figuras 1 a 3 (abaixo) apareceu em uma publicação anterior utilizando este paradigma 8. Figura 4 apareceu em uma tese de doutoramento 36. Estes dados ilustram o impacto da WR-CE sobre a neurogênese adulta hipocampal no...

Watch this Scientific Journal Video about Wheel Running and Environmental Complexity as a Therapeutic Intervention in an Animal Model of FASD at JoVE.com

Wheel Running and Environmental Complexity as a Therapeutic Intervention in an Animal Model of FASD

O exercício cardiovascular e estimulantes experiências em um ambiente complexo ter benefícios positivos em várias medidas de neuroplasticidade dentro do cérebro de roedores. Este artigo irá discutir a implementação destas intervenções como um &quot;superintervention&quot;, que combina corrida roda e complexidade ambiental e abordará as limitações dessas intervenções.

Corrida roda e complexidade ambiental como uma intervenção terapêutica em um modelo animal de FASD

Aerobic exercise (e.g., wheel running (WR) extensively used in animal research) positively impacts many measures of neuroplastic potential in the brain, ...

wheel-running-environmental-complexity-as-therapeutic-intervention-an

Research

JoVE Journal

Behavior

6.7K visualizações.  University of Delaware.  O exercício cardiovascular e estimulantes experiências em um ambiente complexo ter benefícios positivos em várias medidas de neuroplasticidade dentro do cérebro de roedores. Este artigo irá discutir a implementação destas intervenções como um "superintervention", que combina corrida roda e complexidade ambiental e abordará as limitações dessas intervenções. 

Vídeo: Wheel Running and Environmental Complexity as a Therapeutic Intervention in an Animal Model of FASD

The Crossmodal Congruency Task as a Means to Obtain an Objective Behavioral Measure in the Rubber Hand Illusion Paradigm

The rubber hand illusion (RHI) is a popular experimental paradigm. Participants view touch on an artificial rubber hand while the participants' own hidden hand is touched. If the viewed and felt touches are given at the same time then this is sufficient to induce the compelling experience that the rubber hand is one's own hand. The RHI can be used to investigate exactly how the brain constructs distinct body representations for one's own body. Such representations are crucial for successful interactions with the external world. To obtain a subjective measure of the RHI, researchers typically ask participants to rate statements such as "I felt as if the rubber hand were my hand". Here we demonstrate how the crossmodal congruency task can be used to obtain an objective behavioral measure within this paradigm.
The variant of the crossmodal congruency task we employ involves the presentation of tactile targets and visual distractors. Targets and distractors are spatially congruent (i.e. same finger) on some trials and incongruent (i.e. different finger) on others. The difference in performance between incongruent and congruent trials - the crossmodal congruency effect (CCE) - indexes multisensory interactions. Importantly, the CCE is modulated both by viewing a hand as well as the synchrony of viewed and felt touch which are both crucial factors for the RHI.
The use of the crossmodal congruency task within the RHI paradigm has several advantages. It is a simple behavioral measure which can be repeated many times and which can be obtained during the illusion while participants view the artificial hand. Furthermore, this measure is not susceptible to observer and experimenter biases. The combination of the RHI paradigm with the crossmodal congruency task allows in particular for the investigation of multisensory processes which are critical for modulations of body representations as in the RHI.

We demonstrate how an objective measure can be employed in the widely employed rubber hand illusion paradigm. This measure is obtained by modifying the well-established crossmodal congruency task. This task allows the investigation of multisensory processes which are critical for modulations of body representations as in the rubber hand illusion.

A Task Congruência Crossmodal como um meio para se obter uma medida comportamental Objetivo no Rubber Mão Illusion Paradigm

The rubber hand illusion (RHI) is a popular experimental paradigm. Participants view touch on an artificial rubber hand while the participants' own hidden ...

Comportamento

Nest Building as an Indicator of Health and Welfare in Laboratory Mice

The minimization and alleviation of suffering has moral and scientific implications. In order to mitigate this negative experience one must be able to identify when an animal is actually in distress. Pain, illness, or distress cannot be managed if unrecognized. Evaluation of pain or illness typically involves the measurement of physiologic and behavioral indicators which are either invasive or not suitable for large scale assessment. The observation of nesting behavior shows promise as the basis of a species appropriate cage-side assessment tool for recognizing distress in mice. Here we demonstrate the utility of nest building behavior in laboratory mice as an ethologically relevant indicator of welfare. The methods presented can be successfully used to identify thermal stressors, aggressive cages, sickness, and pain. Observation of nest building behavior in mouse colonies provides a refinement to health and well-being assessment on a day to day basis.

We demonstrate the utility of nest building behavior in laboratory mice as an indicator of welfare. Nest scoring is a sensitive technique that is altered by temperature, illness, and aggression. The time to integrate into nest test (TINT) is a simple cage-side assessment that can detect postoperative pain.

The minimization and alleviation of  suffering has moral and scientific implications. In order to mitigate this  negative experience one must be able to ...

The Use of the Puzzle Box as a Means of Assessing the Efficacy of Environmental Enrichment

Environmental enrichment can dramatically influence the development and function of neural circuits. Further, enrichment has been shown to successfully delay the onset of symptoms in models of Huntington&#8217;s disease 1-4, suggesting environmental factors can evoke a neuroprotective effect against the progressive, cellular level damage observed in neurodegenerative disorders. The ways in which an animal can be environmentally enriched, however, can vary considerably. Further, there is no straightforward manner in which the effects of environmental enrichment can be assessed: most methods require either fairly complicated behavioral paradigms and/or postmortem anatomical/physiological analyses. This protocol describes the use of a simple and inexpensive behavioral assay, the Puzzle Box 5-7 as a robust means of determining the efficacy of increased social, sensory and motor stimulation on mice compared to cohorts raised in standard laboratory conditions. This simple problem solving task takes advantage of a rodent&#8217;s innate desire to avoid open enclosures by seeking shelter. Cognitive ability is assessed by adding increasingly complex impediments to the shelter&#8217;s entrance. The time a given subject takes to successfully remove the obstructions and enter the shelter serves as the primary metric for task performance. This method could provide a reliable means of rapidly assessing the efficacy of different enrichment protocols on cognitive function, thus paving the way for systematically determining the role specific environmental factors play in delaying the onset of neurodevelopmental and neurodegenerative disease.

Environmental enrichment provides a potential protective effect against neurodegenerative disorders. Currently, however, there is no easy way of determining the efficacy of enrichment procedures. This protocol describes a simple &#8220;Puzzle Box&#8221; method for assessing an animal&#8217;s cognitive function, in order to reveal the effectiveness of environmental enrichment.

O uso do Puzzle Box como uma forma de avaliar a eficácia de Enriquecimento Ambiental

Environmental enrichment can dramatically influence the development and function of neural circuits.  Further, enrichment has been shown to successfully ...

The Forced Swim Test as a Model of Depressive-like Behavior

The goal of the present protocol is to describe the forced swim test (FST), which is one of the most commonly used assays for the study of depressive-like behavior in rodents. The FST is based on the assumption that when placing an animal in a container filled with water, it will first make efforts to escape but eventually will exhibit immobility that may be considered to reflect a measure of behavioral despair. This test has been extensively used because it involves the exposure of the animals to stress, which was shown to have a role in the tendency for major depression. Additionally, the FST has been shown to share some of the factors that are influenced or altered by depression in humans, including changes in food consumption, sleep abnormalities and drug-withdrawal-induced anhedonia. The main advantages of this procedure are that it is relatively easy to perform and that its results are easily and quickly analyzed. Moreover, its sensitivity to a broad range of antidepressant drugs that makes it a suitable screening test is one of the most important features leading to its high predictive validity. Despite its appeal, this model has a number of disadvantages. First, the issue of chronic augmentation is problematic in this test because in real life patients need to be treated for at least several weeks before they experience any relief from their symptoms. Last, due to the aversiveness of the FST, it is important to take into account possible influences it might have on brain structure/function if brain analyses are to be carried out following this procedure.

This protocol describes the forced swim test, which is used for the study of depressive-like behavior in rodents. This procedure involves placing an animal in a container filled with water that eventually will lead to the exhibition of immobility behavior, which is considered to reflect behavioral despair.

O Teste de Natação Forçada como um modelo de Depressive-like Comportamento

The goal of the present protocol is to describe the forced swim test (FST), which is one of the most commonly used assays for the study of depressive-like ...

An Innovative Running Wheel-based Mechanism for Improved Rat Training Performance

This study presents an animal mobility system, equipped with a positioning running wheel (PRW), as a way to quantify the efficacy of an exercise activity for reducing the severity of the effects of the stroke&#160;in rats. This system provides more effective animal exercise training than commercially available systems such as treadmills and motorized running wheels (MRWs). In contrast to an MRW that can only achieve speeds below 20 m/min, rats are permitted to run at a stable speed of 30 m/min on a more spacious and high-density rubber running track supported by a 15 cm wide acrylic wheel with a diameter of 55 cm in this work. Using a predefined adaptive acceleration curve, the system not only reduces the operator error but also trains the rats to run persistently until a specified intensity is reached. As a way to evaluate the exercise effectiveness, real-time position of a rat is detected by four pairs of infrared sensors deployed on the running wheel. Once an adaptive acceleration curve is initiated using a microcontroller, the data obtained by the infrared sensors are automatically recorded and analyzed in a computer. For comparison purposes, 3 week training is conducted on rats using a treadmill, an MRW and a PRW. After surgically inducing middle cerebral artery occlusion (MCAo), modified neurological severity scores (mNSS) and an inclined plane test were conducted to assess the neurological damages to the rats. PRW is experimentally validated as the most effective among such animal mobility systems. Furthermore, an exercise effectiveness measure, based on rat position analysis, showed that there is a high negative correlation between the effective exercise and the infarct volume, and can be employed to quantify a rat training in any type of brain damage reduction&#160;experiments.

This study presents an innovative running wheel-based animal mobility system to quantify an effective exercise activity in rats. A rat-friendly testbed is built, using a predefined adaptive acceleration curve, and a high correlation between the effective exercise rate and the infarct volume suggests the protocol&#39;s potential for stroke prevention experiments.

Um mecanismo baseado em roda de corrida inovadora para melhorar o desempenho Rat Formação

This study presents an animal mobility system, equipped with a positioning running wheel (PRW), as a way to quantify the efficacy of an exercise activity ...

Introducing Clicker Training as a Cognitive Enrichment for Laboratory Mice

Establishing new refinement strategies in laboratory animal science is a central goal in fulfilling the requirements of Directive 2010/63/EU. Previous research determined a profound impact of gentle handling protocols on the well-being of laboratory mice. By introducing clicker training to the keeping of mice, not only do we promote the amicable treatment of mice, but we also enable them to experience cognitive enrichment. Clicker training is a form of positive reinforcement training using a conditioned secondary reinforcer, the "click" sound of a clicker, which serves as a time bridge between the strengthened behavior and an upcoming reward. The effective implementation of the clicker training protocol with a cohort of 12 BALB/c inbred mice of each sex proved to be uncomplicated. The mice learned rather quickly when challenged with tasks of the clicker training protocol, and almost all trained mice overcame the challenges they were given (100% of female mice and 83% of male mice). This study has identified that clicker training for mice strongly correlates with reduced fear in the mice during human-mice interactions, as shown by reduced anxiety-related behaviors (e.g., defecation, vocalization, and urination) and fewer depression-like behaviors (e.g., floating). By developing a reliable protocol that can be easily integrated into the daily routine of the keeping of laboratory mice, the lifetime experience of welfare in the mice can be improved substantially.

The development of new refinement strategies for laboratory mice is a challenging task that contributes towards fulfilling the 3R principle. This protocol introduces clicker training as a cognitive enrichment program for laboratory mice.

Apresentando Clicker Formação como um enriquecimento cognitivo para Laboratório Mice

Establishing new refinement strategies in laboratory animal science is a central goal in fulfilling the requirements of Directive 2010/63/EU. Previous ...

Establishment of a Valuable Mimic of Alzheimer's Disease in Rat Animal Model by Intracerebroventricular Injection of Composited Amyloid Beta Protein

Alzheimer&#39;s disease (AD) is an irreversible, progressive brain disease that slowly destroys memory and is accompanied by neuron loss and structure change. With the increase of AD patients worldwide, the pathology and treatment of the disease has become a focus in the International Pharmaceutical Industry. Thus, the establishment of the animal model to mimic AD in the laboratory is of great importance.
Here, we describe a detailed protocol for establishing a mimic of AD in a rat animal model though intracerebroventricular injection of amyloid beta protein 25-35 (A&#946;25-35) combined with aluminum trichloride (AlCl3) and anterodorsal thalamic nucleus injection of recombinant human transforming growth factor-&#946;1 (RHTGF-&#946;1) to rats. The related markers of AD were measured, including: spatial memory, neuronal structure and substructure, neuronal A&#946;, and neurofibrillary tangles (NFT) production. This rat model demonstrates spatial memory impairment, neuronal structure and substructure pathological changes, neuron intracellular A&#946; burden, and NFT aggregation, and provides a close mimic of the neuronal structure and function disorder to that of clinical AD patients. Thus, the presented AD rat modelprovides a valuable in vivo tool for exploring neuronal function, neuronal pathology, and drug screening of AD.

This is a protocol to mimic Alzheimer&#39;s Disease in rats by evaluation of spatial memory impairment, neuronal pathological changes, neuronal amyloid beta protein (A&#946;) burden, and neurofibrillary tangles aggregation, induced by the injection of A&#946;25-35 combined with aluminum trichloride and recombinant human transforming growth factor-&#946;1.

Estabelecimento de um mímico valioso de doença de Alzheimer em modelo Animal de rato por injeção Intracerebroventricular de proteína Beta amiloide compostos

Alzheimer&#39;s disease (AD) is an irreversible, progressive brain disease that slowly destroys memory and is accompanied by neuron loss and structure ...

Testing Animal Anxiety in Rats: Effects of Open Arm Ledges and Closed Arm Wall Transparency in Elevated Plus Maze Test

The elevated plus maze test is a behavioral test for assessing animal anxiety in rodents. Although this test is widely applied in the field of behavioral science, conflicting outcomes are often provided from different laboratories. To identify reasons for the different outcomes, we previously focused on arm features, which differ between laboratories, most notably the presence/absence of ledges at the sides of open arms and the transparency/opaqueness of closed arm walls. In a previous report, we used a custom designed container to compare rat behavior on different combinations of open and closed arm designs under otherwise identical experimental conditions, and showed that differences in arm features interfere with experimental outcomes. In brief, open arm ledges significantly increased anxiety-like behavior in rats, while transparent arms may also have decreased this behavior. Furthermore, we verified a higher detection sensitivity of the effect of an anxiolytic drug in a combination of no-ledged open arm + opaque walled closed arm compared with a combination of ledged open arm + transparent walled closed arm. In this report, we introduce our protocol for the elevated plus maze test, together with discussion of the key results from the previous report and our experimental experience. We believe this report will provide useful information for researchers who have employed or who plan to use the elevated plus maze in their studies.

Rats' anxiety-like behavior tested by the elevated plus maze varies highly depending on the testing environment. Here we show how apparatus settings, especially arm structure, affect behavior on the maze, and we explain a reliable protocol to detect effects of anxiolytic drugs. This report will contribute to anxiety research.

Testes Animal ansiedade em ratos: efeitos de bordas de braço aberto e fechado do braço de parede transparência em elevados além de teste de labirinto

The elevated plus maze test is a behavioral test for assessing animal anxiety in rodents. Although this test is widely applied in the field of behavioral ...

Nest Building Behavior as an Early Indicator of Behavioral Deficits in Mice

Nest building is an innate behavior in male and female rodents, even when raised in laboratory settings. As such, many researchers provide rodents synthetic and/or natural materials (such as twine, tissue, cotton, paper, and hay) as a gauge of their overall well-being and as an ancillary assessment to predict the possible decline in cognition. Typically, changes in nesting behaviors, such as failure to create a nest, indicate a change in health or welfare. In addition, nesting behavior is sensitive to many environmental and physiological challenges, as well as many genetic mutations underlying pathological disease states. The following protocol describes a nesting behavior paradigm that explores the usage of four types of nesting material. In addition, the protocol utilizes intraclass correlations to demonstrate that inter-rater reliability is higher when nests are constructed out of shredded paper compared to other common nesting materials such as cotton squares, paper twists, and soft cob bedding. The chosen methodology and statistical considerations (i.e., intraclass correlation) for this assay may be of interest for those conducting experiments assessing the quality of living of mice.&#160;

Here, we present a protocol to quantify nest building behavior in mice, which is known to be impaired in several neurological disorders and diseases. This protocol examines the utility of four materials and offers the opportunity to quantify the rater agreement in scoring, improving the validity and reliability of the assay.

Comportamento do edifício ninho como um indicador precoce de déficits comportamentais em camundongos

Nest building is an innate behavior in male and female rodents, even when raised in laboratory settings. As such, many researchers provide rodents ...

The Use of Traditional Fear Tests to Evaluate Different Emotional Circuits in Cattle

Animal temperament is complex and has implications for productivity and economic profitability. Quantifying an animal&#39;s response to differing stimuli may facilitate breeding selections and identify animals that are better suited to specific management strategies. Multiple tests have been developed to evaluate cattle temperament (e.g., exit velocity, chute score, pen score, open field test, startle test, bovine zero maze), but each of these tests evaluates the animal&#39;s response to different stimuli (e.g., isolation, novel environment, startle, willingness to enter an enclosed area). Cattle temperament has been observed to be relatively stable over time. However, the evaluation of temperament has the potential to be influenced by current conditions, previous experiences, and observer bias. Many of these temperament tests have been improperly categorized as fear tests and have also been criticized for being subjective. This paper provides a framework for standardizing behavioral tests for cattle and suggests that these different evaluations assess different aspects of the animal&#39;s overall temperament.

Here, we present a protocol to conduct a variety of behavioral tests in cattle that have been designed to evaluate emotions. A battery of behavioral tests (open field test, startle test, bovine zero maze, exit velocity, pen score, and chute score) were conducted to evaluate different components of animal temperament.

O uso de testes tradicionais de medo para avaliar diferentes circuitos emocionais em bovinos

Animal temperament is complex and has implications for productivity and economic profitability. Quantifying an animal&#39;s response to differing stimuli ...