This work is funded by the Swiss National Foundation (31_138429) and Synapsis Foundation for the support of research on Alzheimer&#8217;s disease.

Todos os procedimentos com animais estão em conformidade com a directiva comunitária 2010/63 / UE relativa à protecção dos animais utilizados para fins científicos e são aprovados pelo Comitê Animal Care locais (cantão de Fribourg, Suíça). 1. Preparação de animais <ol><li> Os animais experimentais <ol><li> Realizar experimentos sobre tipo selvagem macho adulto e camundongos transgênicos (C57BL / 6 background) de 3-5 meses de idade. Os três grupos de ratos correspondem a controlos de tipo selvagem da mesma ninhada (grupo A, Jagged1 Flox / Flox 19) e duas linhas de ratinhos KO condicionais (grupo B, e C Jagged1ncKO, Jagged1gcKO). </li><li> Os ratos domésticos em condições de laboratório padrão em uma sala ventilada, com um ciclo de 12 hr controlado claro / escuro e fornecer comida e água ad libitum. </li></ol></li></ol> 2. Setup Experimental <ol><li> Arena Experimental <ol><li> Para a arena experimental, use uma gaiola de rato esterilizado limpo (36 cm de comprimento x 20,5 centímetros de largura x 13,5 centímetros de altura) (Figura 1A). </li><li> Atribuir a cada rato para uma gaiola contado com camas frescas, 3 centímetros de altura. Se gaiolas são reutilizados, como no teste de sensibilidade odor, tomar as seguintes medidas para evitar a contaminação cruzada entre odores e camundongos. <ol><li> Marque lado da água. </li><li> Limpar as paredes mais estreitas das gaiolas com dois papéis de seda pulverizados com etanol a 70%, um para cada lado. </li><li> Empilhe gaiolas de acordo com o genótipo dos ratos e armazenar temporariamente sob uma capa laminar. </li></ol></li></ol></li><li> Câmera <ol><li> Montar uma câmara num tripé personalizada com o objectivo em 58 cm a partir do fundo da gaiola (Figura 1A). Fixar a posição do tripé e a gaiola e delimitar com marcas para permitir que a câmara seja centrado sobre a parte superior da gaiola. </li><li> Gravar vídeos em 320 pixels x 240 pixels, 15,08 quadros por segundo como arquivos MOV. </li></ol></li><li> Odores <ol><li> Ressuspender o scents, quando indicado, no solvente em que são solúveis. </li><li> Para o teste de preferência manteiga de amendoim uso. Ressuspender a manteiga de amendoim em óleo de amendoim (10% w / v). </li><li> Para evitar o uso de testes puro 2-metilbutírico (2-Mo) de ácido (98%). </li><li> Para o teste de sensibilidade, use urina feminina da mesma colônia mouse e fundo (C57BL / 6). <ol><li> Por conveniência recolher a urina de 1-2 dias antes do teste olfactivo. Contenha e segure o mouse sob o capô com a barriga acima da grade da gaiola. De acordo com a grelha de gaiola colocar uma placa de petri de plástico para recolher as gotas de urina. </li><li> Recolhe-se o de urina a partir de cada uma das fêmeas em um tubo de 1,5 ml e misturar todas as amostras de urina para normalizar para a variabilidade entre os animais. Armazenar a -20 ° C até à sua utilização. </li><li> No dia da experiência, descongelar a urina e executar quatro diluições em água destilada duas vezes com um factor de diluição de 10 (1:10, 1: 100, 1: 1000, 1: 10.000). </li></ol></li></ol></li></ol>3. Teste Olfativo Nota: Neste protocolo odores foram deliberadamente escolhidas que são percebidos como atractivos fortes (manteiga de amendoim e de urina feminina) ou forte repelente (ácido 2-MB) 15. É importante realizar a preferência e testes de sensibilidade aos odores agradáveis ​​antes do teste de esquiva para eliminar a possibilidade de qualquer interferência com o comportamento olfactiva. No entanto, por uma questão de simplicidade, neste artigo, a preferência ea evasão teste será ambos descritos sob o teste de percepção. Cada sessão comportamental começa com uma fase de habituação. <ol><li> Habituação Fase <ol><li> Colocar o animal na gaiola atribuído limpo e deixá-lo explorar durante 5 minutos (Figura 1B). Uma vez que o ambiente da gaiola experimental é familiar para a gaiola de origem, este curto período de tempo é suficiente para permitir a habituação. </li><li> Se o teste de sensibilidade é concluído em um dia, realizar habituação apenas uma vez antes do application maior do odor diluído. Se o teste de sensibilidade é levada a cabo em dias diferentes, em cada dia é necessária uma fase de habituação de uma nova gaiola limpa. </li></ol></li><li> Teste de Percepção <ol><li> Depois de habituação, activar a câmara imediatamente e de pipeta 60 ul de aroma agradável (manteiga de amendoim) e 60 ul do aroma neutro (água da torneira) sobre as paredes opostas da gaiola em cerca de 10 cm a partir da parte inferior (Figura 1C). </li><li> Deixe o rato explorar os odores durante 2 minutos (Figura 1D). Depois disso, desligue a câmera. </li><li> Neste ponto, prosseguir com o próximo do rato a partir da fase de habituação. Realizar o teste de esquiva exactamente da mesma maneira através da aplicação de 60 ul de odor repelente (ácido 2-MB) e 60 ul de água. </li></ol></li><li> Teste de sensibilidade <ol><li> Avaliar o limiar de captação de ratos macho a concentrações crescentes de urina feminina na seguinte ordem: 1: 10.000; 1: 1000; 1: 100; 1:10 e urina puro. </li><li> Depois de habituação, cada rato para expor a diluição mais elevada pipetados pelo experimentador como anteriormente descrito em 3.2.1. </li><li> Grave o comportamento exploratório de urina contra água, dentro de um prazo de 2 minutos em uma câmara de vídeo. Depois de todas as coortes ratos são testados para a mais alta diluição (1: 10.000), para expor uma concentração mais elevada de urina, como indicado acima. </li></ol></li></ol> 4. Post-hoc de Análise de Dados Nota: Todos os testes comportamentais descritas são processadas post hoc seguindo as instruções de análise de dados. <ol><li> Abrir arquivos MOV em ImageJ para sistemas Windows <ol><li> Instale Quick Time for Java usando as configurações personalizadas de http://www.apple.com/quicktime/download. </li><li> Instale o plugin do Quick Time do site da ImageJ (http://rsb.info.nih.gov/ij/plugins/qt-capture.html). </li><li> Importar o QTJava.zip (C: Program Files QuickTime QTSystem) nas extens bibliotecaion de ImageJ (.ImageJ jre lib ext). </li><li> Copie também o QTJava.zip na pasta plugins e renomeá-lo como QTJava.jar. </li><li> Instale as seis scripts anexados na pasta Macros (ImageJ plugins Macros). </li><li> Abrir ImageJ e compilar e executar o plugin do Quick Time, a partir daí perto ImageJ. </li><li> Reabrir ImageJ e abra o arquivo MOV usando Arquivo&gt; Importar&gt; usando Quick Time. </li></ol></li><li> Ajuste de Vídeo <ol><li> Uma vez que o ficheiro de vídeo é aberto no ImageJ, cortar o vídeo a fim de obter uma constante de 2 minutos a partir do tempo de exploração do experimentador pipetados os odorantes na gaiola (T0). Identificar o quadro correspondente ao T0 e remover os quadros anteriores usando incrementos de 1 (ImageJ Imagem Stacks ToolsSlice removedor). Use o mesmo comando para excluir todos os quadros superiores a 2 min exploração. </li><li> Certifique-se de que a gaiola é centrado e se necessário, use a Imagem&gt; Transform&gt; comando Girar para alinhá-lo. </li></ol></li><li> Video Processamento Nota: O processamento de vídeo é totalmente assistida por computador e utiliza comandos macros que acompanham este papel. <ol><li> A fim de diminuir a superfície para a gaiola de um 127 pixels x 218 pixels tamanho executar o Passo 1 macro do Plugin&gt; Macros&gt; Executar comando. Mover o rectângulo fixado sobre a gaiola (Figura 2, Passo 1). </li><li> Cortar a área da gaiola sobre a região de interesse (ROI), utilizando o Passo 2 macro (Figura 2, Passo 2). </li><li> Use a macro Passo 3 para extrair a imagem do rato do fundo, atribuindo um sinal de limiar, despeckling e filtrando a variação do sinal. Os valores de saída da parcela do eixo Z indicam os valores de cinzentos médios, correspondendo à intensidade da sombra rato mover-se dentro da ROI da &quot;câmara de água&quot; durante 2 min a exploração. Copie os resultados em uma planilha nomeada de acordo com o ROI em um arquivo de planilha (Figura 2, Passo 3). </li><li> Use a macro Passo 4 paraextrair os valores de cinza médios do rato na ROI &quot;câmara de odor&quot;. Copie os resultados em uma planilha nomeada de acordo com o ROI no mesmo arquivo de planilha como em 4.3.3 (Figura 2, Passo 4). </li><li> Para restringir ainda mais a análise do movimento do mouse no ROI &quot;perímetro água&quot; usar o Passo 5 macro. Copie o resultado na planilha nomeada de acordo com o ROI no mesmo arquivo de planilha como em 4.3.3 (Figura 2, Passo 5). </li><li> Para restringir a análise do movimento do mouse no ROI &quot;perímetro odor&quot; usar a macro Passo 6. Copie o resultado na planilha nomeada de acordo com o arquivo de ROI planilha como em 4.3.3 (Figura 2, Passo 6). </li><li> Processar todos os vídeos e verificar a consistência do número de quadros por animais. Aqui, gravar todos os animais para 1.810 quadros correspondentes a uma sessão de exploração 2 min. </li><li> Para cada animal e para cada ROI tipo quadros com va cinzento médiolues maior do que 0. Divide o número de quadros pelos valores correspondentes a 1 segundo e obter os segundos gastos em cada ROI. </li></ol></li></ol> 5. Análise Estatística <ol><li> Para cada ensaio, verificar homogeneidade da variância dentro de grupos / genótipos pelo teste de Bartlett utilizando a fórmula disponível no http://www.real-statistics.com/one-way-analysis-of-variance-anova/homogeneity-variances/. </li><li> No teste de atração e de evasão, proceder a comparações entre os tempos passados ​​com água contra odor dentro de um grupo usando um teste t de Student não-direcional assumindo variâncias iguais ou desiguais, dependendo dos resultados de teste de Bartlett. Compare os tempos passados ​​com os odores subtraídos pelo tempo gasto com água entre os genótipos por ANOVA one-way com teste post-hoc de Bonferroni. </li><li> No teste de sensibilidade analisar as comparações entre o tempo gasto com o odor subtraído do tempo gasto com water entre os grupos em diluições específicas de urina por ANOVA one-way com teste post-hoc de Bonferroni. Comparar a sensibilidade entre grupos às crescentes concentrações de odor por 2-Way ANOVA com repetições com teste post-hoc de Bonferroni. </li><li> Interação entre genótipos e tratamentos no teste de atração e evasão são investigados por 2-way ANOVA com o teste post-hoc de Bonferroni. </li></ol>

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There is no conflict of interest.

Os testes propostos neste protocolo permitem avaliar diferentes aspectos do comportamento olfativo inata em camundongos: a percepção de odores, uma discriminação entre os odores contra água e sensibilidade aos odores. Este protocolo pode ser aplicado a qualquer odor de acordo com a preferência e escala evitar mostrado previamente 15. Uma vez que o protocolo baseia-se na actividade exploratória é importante que os ratinhos não exibem qualquer deficiência motora ou ansiedade que podem afectar o seu mo...

Olfaction é uma das funções sensoriais e mais desenvolvidos importantes em mamíferos. Qualquer prejuízo na atividade olfativa pode afetar a ingestão de alimentos, comportamento social e, no pior dos casos, até mesmo a sobrevivência. Nos seres humanos, a deterioração olfactiva é dependente 1 idade e é considerado um forte indicador de distúrbios neurológicos 2-6. O teste de identificação olfatória desenvolvido pela Universidade da Pensilvânia atualmente representa um dos testes mais utilizados, não-invasivos e quantificáveis, de diagnóstico, que pode avaliar primeiros déficits neurológicos 7 e prever com elevada probabilidade, a progressão da demência 8,9. A acessibilidade do sistema olfativo ea proeminência do olfato em roedores, provocou uma intensa linha de pesquisa abordando os mecanismos subjacentes funções olfativas 10. Nós já demonstrado que a perda de função do recept sinalizaçãoou Notch1 afeta evitar olfativo 11. Neste protocolo, usamos ratos que faltam o ligando sinalização, Jagged1, em neurônios ou células gliais para estudar o desempenho olfativo. Olfaction inata é definido por três parâmetros como percepção, discriminação entre odores e sensibilidade olfativa 4. Teste olfativo em roedores pode ser feito de uma variedade de formas e alguns estudos comportamentais fazer uso de olfatômetros, que fornecem o odor para o animal a uma concentração de vapores específicos e num período de tempo preciso 12-14. No entanto, esta instrumentação é caro e pode estar disponíveis apenas em instalações especializadas. No nosso trabalho, nós fornecemos um protocolo de teste olfactivo simples, rápido e reprodutível, que é levada a cabo usando aromas voláteis. Os testes descritos percepção medida para um atrativo ou um odor repelente e avaliar a discriminação entre o cheiro eo 11,15,16 água. Usando a mesma configuração, we também se pode medir a sensibilidade a um odor a diferentes concentrações 16,17. O processamento de vídeo assistida por computador post-hoc, inspirada no trabalho da página e colegas 18, fornece resultados imparciais, sem a necessidade de cegueira experimental e permitindo para uma única pessoa para levar a cabo todo o experimento. Este protocolo destina-se a fornecer um ponto de partida para estudar o comportamento olfativo em camundongos.

<table border="0" cellpadding="0" cellspacing="0" style="border-collapse:
 collapse;width:192pt">
	<colgroup>
		<col span="4" />
	</colgroup>
	<tbody>
		<tr>
			<td class="xl63">Mouse cage</td>
			<td class="xl64">Italplast (Italy)</td>
			<td class="xl64">1144B</td>
			<td class="xl64">36 cm length x 20.5 cm width x 13.5 cm height</td>
		</tr>
		<tr>
			<td class="xl63">Chipped wood bedding</td>
			<td class="xl64">Abedd (Austria)</td>
			<td class="xl64">LTE E-001</td>
			<td class="xl64">3 cm high</td>
		</tr>
		<tr>
			<td class="xl63">Peanut butter</td>
			<td class="xl64">Migros (Swizterland)</td>
			<td class="xl64">NA</td>
			<td align="right" class="xl65">1:10</td>
		</tr>
		<tr>
			<td class="xl63">2-Methylbutyric</td>
			<td class="xl64">Sigma Aldrich (Switzerland)</td>
			<td class="xl64">W269514</td>
			<td class="xl64">Pure</td>
		</tr>
		<tr>
			<td class="xl63">Female Urine from fertile females of same mouse strain</td>
			<td class="xl64">NA</td>
			<td class="xl64">NA</td>
			<td class="xl64">Dilution series</td>
		</tr>
		<tr>
			<td class="xl63">Camera</td>
			<td class="xl64">Olympus (US)</td>
			<td class="xl64">Camedia C-8080</td>
			<td class="xl64">MOV files</td>
		</tr>
		<tr>
			<td class="xl63">Quicktime for Java (Windows)</td>
			<td class="xl64">Apple (USA)</td>
			<td class="xl64">NA</td>
			<td class="xl64">video plugin for visualizing MOV files</td>
		</tr>
		<tr>
			<td class="xl63">Image J for Windows</td>
			<td class="xl64">NIH (USA)</td>
			<td class="xl64">NA</td>
			<td class="xl64">Video Processing/Analysis</td>
		</tr>
	</tbody>
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simple and computer-assisted olfactory testing for mice

Olfaction é altamente conservada entre espécies e é necessária para a reprodução e sobrevivência. Nos seres humanos, olfacto é também um dos sentidos que é afectado com o envelhecimento e é um forte indicador de doenças neurodegenerativas. Assim, o teste de olfacto é utilizado como um método de diagnóstico não invasivo para detectar os défices neurológicos no início. A fim de compreender os mecanismos subjacentes susceptibilidade rede olfativo, a investigação olfativa em roedores ganhou impulso na última década. Aqui, apresentamos um tempo método muito simples, eficiente e reproduzível teste olfativo de percepção de odor inata e sensibilidade em ratos sem a necessidade de qualquer restrição de alimentos ou água antes. Os testes são realizados em um ambiente familiar para os ratos, requerem apenas os aromas e uma sessão de 2 min de exposição odorante. A análise é realizada, post-hoc, usando comandos assistidas por computador em ImageJ e pode ser, portanto,, Realizado do início ao fim por um pesquisador. Este protocolo não requer nenhum hardware ou configuração especial e é indicado para qualquer laboratório interessado em testar percepção olfativa e sensibilidade.

O teste mede a percepção atração a manteiga de amendoim e evasão de ácido 2-MB. Três grupos de ratinhos são testados e o tempo gasto no &quot;perímetro odor&quot; são quantificadas em relação à água. No teste de preferência, o grupo controlo exibe uma significativa preferência para o odor em comparação com a água (8 t = 2,52, p &lt;0,05). Por outro lado, o grupo B não apresenta qualquer atracção significativa para a manteiga de amendoim e gasta mais tempo com água (6 t = 3,22...

Watch this Scientific Journal Video about Simple and Computer-assisted Olfactory Testing for Mice at JoVE.com

Simple and Computer-assisted Olfactory Testing for Mice

We present a simple and unbiased olfactory test in mice. With this protocol olfactory discrimination, preference, avoidance and sensitivity to a novel odor as compared to water can be assessed in single behavioral sessions. This method is indicated for a single experimenter and analysis is based on computer-assisted video processing.

Simples e Testes Olfativo para Ratos assistido por computador

Olfaction is highly conserved among species and is required for reproduction and survival.
In humans, olfaction is also one of the senses that is affected ...

simple-and-computer-assisted-olfactory-testing-for-mice

Research

JoVE Journal

Behavior

9.9K visualizações.  University of Fribourg. We present a simple and unbiased olfactory test in mice. With this protocol olfactory discrimination, preference, avoidance and sensitivity to a novel odor as compared to water can be assessed in single behavioral sessions. This method is indicated for a single experimenter and analysis is based on computer-assisted video processing. 

Vídeo: Simple and Computer-assisted Olfactory Testing for Mice

Barnes Maze Testing Strategies with Small and Large Rodent Models

Spatial learning and memory of laboratory rodents is often assessed via navigational ability in mazes, most popular of which are the water and dry-land (Barnes) mazes. Improved performance over sessions or trials is thought to reflect learning and memory of the escape cage/platform location. Considered less stressful than water mazes, the Barnes maze is a relatively simple design of a circular platform top with several holes equally spaced around the perimeter edge. All but one of the holes are false-bottomed or blind-ending, while one leads to an escape cage. Mildly aversive stimuli (e.g.&#160;bright overhead lights) provide motivation to locate the escape cage. Latency to locate the escape cage can be measured during the session; however, additional endpoints typically require video recording. From those video recordings, use of automated tracking software can generate a variety of endpoints that are similar to those produced in water mazes (e.g.&#160;distance traveled, velocity/speed, time spent in the correct quadrant, time spent moving/resting, and confirmation of latency). Type of search strategy (i.e.&#160;random, serial, or direct) can be categorized as well. Barnes maze construction and testing methodologies can differ for small rodents, such as mice, and large rodents, such as rats. For example, while extra-maze cues are effective for rats, smaller wild rodents may require intra-maze cues with a visual barrier around the maze. Appropriate stimuli must be identified which motivate the rodent to locate the escape cage. Both Barnes and water mazes can be time consuming as 4-7 test trials are typically required to detect improved learning and memory performance (e.g.&#160;shorter latencies or path lengths to locate the escape platform or cage) and/or differences between experimental groups. Even so, the Barnes maze is a widely employed behavioral assessment measuring spatial navigational abilities and their potential disruption by genetic, neurobehavioral manipulations, or drug/ toxicant exposure.

The dry-land Barnes maze is widely used to measure spatial navigation ability in response to mildly aversive stimuli. Over consecutive days, performance (e.g.&#160;latency to locate escape cage) of control subjects improves, indicative of normal learning and memory. Differences between rats and mice necessitate apparatus and methodology changes that are detailed here.

Barnes Estratégias Teste Labirinto com pequenas e grandes roedores Modelos

Spatial learning and memory of laboratory rodents is often assessed via navigational ability in mazes, most popular of which are the water and dry-land ...

Comportamento

Testing for Odor Discrimination and Habituation in Mice

This video demonstrates a technique to establish the presence of a normally functioning olfactory system in a mouse. The test helps determine whether the mouse can discriminate between non-social odors and social odors, whether the mouse habituates to a repeatedly presented odor, and whether the mouse demonstrates dishabituation when presented with a novel odor. Since many social behavior tests measure the experimental animal&#8217;s response to a familiar or novel mouse, false positives can be avoided by establishing that the animals can detect and discriminate between social odors. There are similar considerations in learning tests such as fear conditioning that use odor to create a novel environment or olfactory cues as an associative stimulus. Deficits in the olfactory system would impair the ability to distinguish between contexts and to form an association with an olfactory cue during fear conditioning. 
In the odor habitation/dishabituation test, the mouse is repeatedly presented with several odors. Each odor is presented three times for two minutes. The investigator records the sniffing time directed towards the odor as the measurement of olfactory responsiveness. A typical mouse shows a decrease in response to the odor over repeated presentations (habituation). The experimenter then presents a novel odor that elicits increased sniffing towards the new odor (dishabituation). After repeated presentation of the novel odor the animal again shows habituation. This protocol involves the presentation of water, two or more non-social odors, and two social odors. In addition to reducing experimental confounds, this test can provide information on the function of the olfactory systems of new knockout, knock-in, and conditional knockout mouse lines.

This manuscript describes a protocol to examine the olfactory system of rodents. The olfactory habituation/dishabituation test will allow investigators to determine whether a mouse habituates to a repeatedly presented odor and whether the mouse demonstrates dishabituation when presented a novel odor.

Testes para Odor Discriminação e habituação em ratos

This video demonstrates a technique to establish the presence of a normally functioning olfactory system in a mouse. The test helps determine whether the ...

Morris Water Maze Test: Optimization for Mouse Strain and Testing Environment

The Morris water maze (MWM) is a commonly used task to assess hippocampal-dependent spatial learning and memory in transgenic mouse models of disease, including neurocognitive disorders such as Alzheimer&#8217;s disease. However, the background strain of the mouse model used can have a substantial effect on the observed behavioral phenotype, with some strains exhibiting superior learning ability relative to others. To ensure differences between transgene negative and transgene positive mice can be detected, identification of a training procedure sensitive to the background strain is essential. Failure to tailor the MWM protocol to the background strain of the mouse model may lead to under- or over- training, thereby masking group differences in probe trials. Here, a MWM protocol tailored for use with the F1 FVB/N x 129S6 background is described. This is a frequently used background strain to study the age-dependent effects of mutant P301L tau (rTg(TauP301L)4510 mice) on the memory deficits associated with Alzheimer&#8217;s disease. Also described is a strategy to re-optimize, as dictated by the particular testing environment utilized.

This manuscript describes a Morris water maze (MWM) protocol tailored for use with a commonly used mouse model of Alzheimer's disease. The MWM is widely used in transgenic mouse models. Implementation of a procedure sensitive to the background strain of the mouse model is essential for detecting group differences.

Morris Water Maze Teste: Otimização para Mouse e Strain Testing Ambiente

 The Morris water maze (MWM) is a commonly used task to assess hippocampal-dependent spatial learning and memory in transgenic mouse models of disease, ...

The Treadmill Fatigue Test: A Simple, High-throughput Assay of Fatigue-like Behavior for the Mouse

Fatigue is a prominent symptom in many diseases and disorders and reduces quality of life for many people. The lack of clear pathogenesis and failure of current interventions to adequately treat fatigue in all patients leaves a need for new treatment options. Despite the therapeutic need and importance of preclinical research in helping identify promising novel treatments, few preclinical assays of fatigue are available. Moreover, the most common preclinical assay used to assess fatigue-like behavior, voluntary wheel running, is not suitable for use with some strains of mice, may not be sensitive to drugs that reduce fatigue, and has relatively low throughput. The current protocol describes a novel, non-voluntary preclinical assay of fatigue-like behavior, the treadmill fatigue test, and provides evidence of its efficacy in detecting fatigue-like behavior in mice treated with a chemotherapy drug known to cause fatigue in humans and fatigue-like behavior in animals. This assay may be a beneficial alternative to wheel running, as fatigue-like behavior and potential interventions can be assessed in a greater number of mice over a shorter time frame, thus permitting faster discovery of new therapeutic options.

Fatigue is a common, undertreated and frequently poorly-understood symptom in many diseases and disorders. New preclinical assays of fatigue may help to improve current understanding and future treatment of fatigue. To that end, the current protocol provides a novel means of measuring fatigue-like behavior in the mouse.

A esteira de fadiga de teste: Um ensaio simples, de alta taxa de transferência de Fadiga-like Comportamento para o Mouse

Fatigue is a prominent symptom in many diseases and disorders and reduces quality of life for many people. The lack of clear pathogenesis and failure of ...

Task Interruption and Resumption Paradigm for Testing the Activation and Pursuit of an Abstract Thinking Goal

This protocol is based on the task interruption and resumption paradigm, the premise of which is that active goals lead to persistent behavior and thus a higher resumption rate after a period of delay or interruption. The task interruption and resumption protocol described in this research is tailored to test the activation of cognitive goals (e.g., a goal to think more abstractly). Cognitive goals may be pursued even during the interruption period; thus, to prevent this, the protocol involves cognitive distraction. The protocol consists of several stages. Specifically, the initial stage includes the goal activation process, where the treatment (versus control) condition receives a manipulation to activate the cognitive goal being tested by the researcher. In the next stage, participants are presented with the introduction of a task that is perceived to either satisfy or not satisfy the cognitive goal of interest. Importantly, this task is interrupted a few seconds after it begins. The task interruption forces a delay period and introduces a cognitive distraction to prevent the automatic pursuit and fulfillment of the cognitive goal. After the interruption period, participants are given a choice between resuming the interrupted task and abandoning the interrupted task to complete an alternative task instead. Among participants whose cognitive goals had been activated at the earlier stage, the task resumption rate should be higher if the task was perceived as an opportunity to satisfy (versus not satisfy) the goal. Such a finding would provide empirical evidence that the cognitive goal has been activated and pursued. In previous research, this protocol has been used to test whether causal uncertainty activates an abstract thinking goal. Adapting the protocol to test the activation of other cognitive goals is also discussed.

This protocol was designed to test the activation and pursuit of cognitive goals (e.g., an abstract thinking goal) using the task interruption and resumption paradigm. The protocol is suitable for cognitive goals that are automatically pursued once activated, as the distraction procedure prevents goal pursuit during the interruption period.

Interrupção de tarefas e Paradigm Reinício para testar a Ativação e busca de um objetivo pensamento abstrato

This protocol is based on the task interruption and resumption paradigm, the premise of which is that active goals lead to persistent behavior and thus a ...

A Simple and Low-cost Assay for Measuring Ambulation in Mouse Models of Muscular Dystrophy

Measuring functional outcomes in the treatment of muscular dystrophy is an essential aspect of preclinical testing. The assessment of voluntary ambulation in mouse models is a non-invasive and reproducible activity assay that is directly analogous to measures of patient ambulation such as the 6-minute walk test and related mobility scores. Many common methods for testing mouse ambulation speed and distance are based on the open field test, where an animal&#39;s free movement within an arena is measured over time. One major downside to this approach is that commercial software and equipment for high-resolution motion tracking is expensive and may require transferring mice to specialized facilities for testing. Here, we describe a low-cost, video-based system for measuring mouse ambulation that utilizes free and open-source software. Using this protocol, we demonstrate that voluntary ambulation in the dystrophin-null mdx mouse model for Duchenne muscular dystrophy (DMD) is decreased relative to wild-type mouse activity. In mdx mice expressing the utrophin transgene, these activity deficits are not observed and the total distance traveled is indistinguishable from wild-type mice. This method is effective for measuring changes in voluntary ambulation associated with dystrophic pathology, and provides a versatile platform that can be readily adapted to diverse research settings.

This protocol describes a flexible, low-cost system for measuring mouse ambulation in an open field activity assay. We show that a 6-minute ambulation assay based on this system detects a decrease in voluntary movement in mdx mice, and accurately distinguishes improvement in a muscle-specific rescue of these animals.

Um ensaio simples e de baixo custo para medir a deambulação em modelos de Mouse de Distrofia Muscular

Measuring functional outcomes in the treatment of muscular dystrophy is an essential aspect of preclinical testing. The assessment of voluntary ambulation ...

A Simple Non-invasive Method for Temporary Knockdown of Upper Limb Proprioception

Proprioception may be the least well measured of all contributors to the neural control of movement. New precise, reliable measures of proprioception are needed for clinical diagnosis of impairment, and to measure outcomes of proprioceptive training. The purpose of this simple, non-invasive method is to temporarily knockdown upper limb proprioception in healthy adults, to an extent that would be useful in the development and testing of upper limb proprioception measures. Knockdown models have two main advantages over studying humans with impaired proprioception: participant availability and the ability to control the extent of impairment across participants. Current published methods of temporary proprioception knockdown of the upper limb, such as ischemic nerve blocks and cryotherapy, are invasive, impractical, or uncomfortable for the participant. Here, vibration over the ulnar groove was used to reduce upper limb proprioception. High frequency vibration may reduce proprioceptive acuity by inhibiting pacinian corpuscle-induced input. The effect of vibration used in this protocol was confirmed using two quantitative measures. This method was simple to administer, comfortable for participants, and practical.

The goal of this protocol is to demonstrate a practical method to temporarily interfere with proprioception in the upper limb of healthy humans.

Um método não-invasivo simples para Knockdown temporária de propriocepção do membro superior

Proprioception may be the least well measured of all contributors to the neural control of movement. New precise, reliable measures of proprioception are ...

Measuring Active and Passive Tameness Separately in Mice

Domesticated animals such as dogs and laboratory mice show a high level of tameness, which is important for humans to handle them easily. Tameness has two behavioral components: a reluctance to avoid humans (passive tameness) and a motivation to approach humans (active tameness). To quantify these components in mice, we previously developed behavioral tests for active tameness, passive tameness, and the willingness to stay on a human hand, each designed to be completed within 3 min. The data obtained were used for selective breeding, with a large number of mice analyzed per generation. The active tameness test measures the movement of the mouse toward a human hand and the contact it engages in. The passive tameness test measures the duration of time that a mouse tolerates human touch. In the stay-on-hand test, a mouse is placed on a human hand and touched slowly using the thumb of that hand; the duration of time that the animal remains on the hand is measured. Here, we describe the test set-up and apparatus, explain the procedures, and discuss the appropriate data analysis. Finally, we explain how to interpret the results.

Tameness in animals includes a reduction of avoidance responses to humans (passive tameness) and an increase in active approaches to humans (active tameness). Here, we describe detailed protocols for three behavioral tests (active tameness, passive tameness, and stay-on-hand tests) to separately measure the active and passive tameness of mice.

Medição de ativa e passiva domesticidade separadamente em ratos

Domesticated animals such as dogs and laboratory mice show a high level of tameness, which is important for humans to handle them easily. Tameness has two ...

Computerized Adaptive Testing System of Functional Assessment of Stroke

The computerized adaptive testing system of the functional assessment of stroke (CAT-FAS) can simultaneously assess four functions (motor functions of the upper and lower extremities, postural control, and basic activities of daily living) with sufficient reliability and administrative efficiency. CAT, a modern measurement method, aims to provide a reliable estimate of the examinee's level of function rapidly. CAT administers only a few items whose item difficulties match an examinee's level of function and, thus, the administered items of CAT can provide sufficient information to reliably estimate the examinee's level of function in a short time. The CAT-FAS was developed through four steps: (1) determining the item bank, (2) determining the stopping rules, (3) validating the CAT-FAS, and (4) establishing a platform of online administration. The results of this study indicate that the CAT-FAS has sufficient administrative efficiency (average number of items = 8.5) and reliability (group-level Rasch reliability: 0.88 - 0.93; individual-level Rasch reliability: &#8805;70% of patients had Rasch reliability score &#8805;0.90) to simultaneously assess four functions in patients with stroke. In addition, because the CAT-FAS is a computer-based test, the CAT-FAS has three additional advantages: the automatic calculation of scores, the immediate storage of data, and the easy exporting of data. These advantages of the CAT-FAS will be beneficial to data management for clinicians and researchers.

Here, we present a protocol to develop the computerized adaptive testing system of the functional assessment of stroke (CAT-FAS). The CAT-FAS can simultaneously assess four functions (two motor functions [upper and lower extremities], postural control, and basic activities of daily living) with sufficient reliability and administrative efficiency.

Sistema de teste adaptativo computadorizado de avaliação funcional do curso

The computerized adaptive testing system of the functional assessment of stroke (CAT-FAS) can simultaneously assess four functions (motor functions of the ...

Olfactory Context Dependent Memory: Direct Presentation of Odorants

Information is retrieved more effectively when the retrieval occurs in the same context as that in which the information was first encoded. This is termed context dependent memory (CDM). One category of cues that have been shown to effectively produce CDM effects are odors. However, it is unclear what the boundary conditions of these CDM effects are. In particular, given that olfaction has been called an implicit sense, it is possible that odors are only effective mnemonic cues when they are presented in the background. This assertion seems even more likely given that previous research has shown odors to be poor cues during paired associate memory tests, where odors are in the focus of attention as mnemonic cues for other information. In order to determine whether odors are only effective contextual mnemonic cues when presented outside the central focus of an observer, an olfactory CDM experiment was performed in which odorants were presented directly, rather than ambiently. Direct presentation was accomplished with the aid of an olfactometer. The olfactometer not only allows for direct presentation of odorants, but provides other methodological benefits, including the allowance of trial by trial manipulations of odorant presentations and, relatedly, time-specific releases of odorants. The presence of the same odor during both encoding and retrieval enhanced memory performance, regardless of whether the odor was presented ambiently or directly. This finding can serve as a basis for future olfactory CDM research which can utilize the benefits of direct presentation.

The use of an olfactometer for directly presenting odorants opens exciting opportunities for researchers of olfactory memory. The current paper discusses issues related to this methodology as related to a previously published experiment on olfactory context dependent memory.

Memória dependente de contexto olfativa: Apresentação directa de odorantes

Information is retrieved more effectively when the retrieval occurs in the same context as that in which the information was first encoded. This is termed ...