Apoio: Esta pesquisa foi apoiada pelo National Institutes of Health (DK064862, NS058525 e AA019357 a GGF).

1. Curto prazo LMA Set Up and Procedure <ol><li> Curto prazo LMA gaiola configurar <ol><li> Teste LMA curto prazo dentro da gaiola de um rato alojados individualmente. Para facilitar a análise de imagens VTS-dependente, usar roupa de cama em alto contraste com a cor do mouse (ou seja, roupa de cama de cor escura para ratos brancos e luz de cama colorida para ratos pretos). </li><li> Durante os testes de LMA de curto prazo, cobrir gaiolas com ¼ claro em acrílico com 11 x ¼ em furos de diâmetro para suportar a ventilação adequada. </li></ol></li><li> Curto prazo sala de testes LMA <ol><li> Teste ratinhos num ambiente limpo, em que tenham sido aclimatados durante pelo menos a 24 horas 8. </li><li> Para iluminar o ambiente de teste, use chocadeira luzes braçadeira com lâmpadas vermelhas (&lt;125 lumens). Antes de videografia, ajustar o ângulo ea posição das luzes para remover o brilho ou reflexos dos topos de gaiola de acrílico. </li><li> Remover ou evitar o ruído ambienteacima de 70 dB. Use um gerador de ruído branco se o ruído de fundo intermitente é inevitável durante o teste de mouse. </li><li> Cubra a superfície de teste no qual as gaiolas são colocadas na sala de testes com o açougueiro ou laboratório de banco de papel de cor semelhante à da cama gaiola, caso o chão da gaiola é exposto durante videografia. </li></ol></li><li> Curto prazo LMA videografia <ol><li> Centro e coloque a noite câmera de vídeo com capacidade de visão do tecto com pelo menos 120 GB de memória acima da área de gravação na sala de testes. Não exceda a capacidade de resolução dos VTS usados, colocando muitas gaiolas dentro da câmera quadro de visualização de vídeo (por exemplo, Ethovision XT 7 por Noldus Tecnologia da Informação (Leesburg VA) pode rastrear simultaneamente oito camundongos em tempo real). </li></ol></li><li> Procedimento de teste LMA curto prazo <ol><li> Teste LMA curto prazo durante a fase ativa / escuro do ciclo de luz. Certifique-se o tempo em relação ao ciclo de luz é the mesmo para todos os grupos, como LMA segue um padrão crepuscular, com mais actividade durante os períodos de início e depois do ciclo de escuro 1. Incluir tanto os sujeitos experimentais e controle em cada grupo de camundongos gravadas para garantir os potenciais efeitos das diferenças temporais na LMA são minimizados. </li><li> Individualmente casas e aclimatar os ratos, pelo menos, 24 horas antes do início do teste de LMA 8. </li><li> Coloque as divisórias de gaiolas e jaulas (contendo camundongos individuais) diretamente sob a câmera. Remova todo o material de nidificação / enriquecimento das gaiolas, pois isso pode obstruir a visão do mouse (por exemplo, se o mouse tocas / cobre-se com o material do assentamento) e interferir com VTS precisão. Coloque uma tampa de acrílico claro em cada gaiola gravado. </li><li> Quando videoing várias gaiolas ao mesmo tempo, certifique-se de que a resolução é compatível com os VTS escolhidos. Para facilitar a análise VTS, posicione as gaiolas de forma idêntica em cada um divisor (ou seja, canto superior direito). </li><li> Iniciar a gravação. Comece cada vídeo com uma lousa que identifica todas as informações experimental relevante para assegurar a identificação correcta do vídeo independente de qualquer documentação não-vídeo adicional. Use um temporizador ou cronômetro para acompanhar o tempo de LMA testes (5-10 min). Se a informação de tempo é perdida, recuperá-lo a partir do registro de vídeo. </li><li> Para as medidas repetidas usando a curto prazo método LMA, permitir ratos acesso a comida e água durante os intervalos de medição não. </li><li> Durante o teste, manter todos os experimentadores fora da vista dos ratos de modo a não agir como um sinal visual externo. </li><li> Quando o teste estiver concluído, o retorno / substitua o material do assentamento / enriquecimento, para reduzir o potencial de desenvolvimento da ansiedade-como o comportamento 8. </li></ol></li><li> LMA análise de curto prazo <ol><li> Armazenar as gravações de vídeo da sessão de testes para uma mídia de arquivamento compatíveis com os VTS usados. Para Ethovision XT 7, uma unidade de disco rígido (HDD) é usado. </li><li> Reproduzir os vídeos no VTS usando seu player de mídia associada, analisando os vídeos em tempo real. Na Figura 1, as variáveis ​​de detecção eram: 2 cm / seg começa rastreamento e 1,75 cm / s rastreamento pára. </li><li> Inicialmente, escolher todo o chão da gaiola, como a região de interesse (ROI) para o rastreamento. Para as análises avançadas, tais como a avaliação de tigmotatismo, criar várias ROIs intra-gaiola. </li><li> Para os estudos de alto rendimento, analisar todas as gaiolas no vídeo simultaneamente com piso ROIs all-gaiola individual. </li><li> Usar o VTS para calcular a distância total movida (cm), a velocidade de movimento (cm / seg) e da duração do movimento (seg). Para Ethovision XT 7, exportar os dados gerados em Microsoft Excel para armazenamento e análise adicional. </li></ol></li></ol> 2. Teste LMA longo prazo Configurar e Procedimento <ol><li> LMA testes de construção de gaiola de longo prazo <ol><li> Corte um buraco de diâmetro 5,5 centímetros na parede de curto-endde uma gaiola padrão mouse. Criar uma cobertura para o buraco com ¼ de malha de aço galvanizado. Corte a malha, criando vários ½ nas aberturas para permitir o acesso do rato à alimentação. Polir qualquer malha corte para remover bordas afiadas. </li><li> Use malha adicional para criar uma tremonha externa directamente ligada à malha utilizada para cobrir a abertura de 5,5 cm. Conecte o funil para a malha usada para cobrir o buraco, e anexar a montagem da gaiola usando rebites de alumínio. </li><li> Construir uma garrafa de água a partir de um padrão de 50 ml cônico (Corning) e uma rolha de borracha 1-hole (Fisher), que é o excesso de perfuração para permitir a inserção de não-gotejamento tubo sipper (Sistemas de Sta-Pure). Fixe a garrafa de água para a gaiola com fita de velcro (3M) e inserir o tubo sipper através da abertura coberta mesh. </li><li> Durante os testes de LMA de longo prazo, cobrir gaiolas com ¼ claro em acrílico com 11 x ¼ em furos de diâmetro para suportar a ventilação adequada. </li></ol></li><li> Longo prazo sala de testes LMA<ol><li> Longo prazo sala de testes LMA criado é idêntico ao LMA curto prazo. Consulte a seção 1.2 para procedimento. </li></ol></li><li> Longo prazo LMA videografia <ol><li> Center e colocar um tecto montado no infravermelho, de estilo de segurança, a câmara directamente ligado a um PC por cima da área da gravação na sala de teste. Apenas registrar o número de gaiolas de uma vez que resulta em resolução gaiola indivíduo dentro da capacidade dos VTS utilizados (por exemplo, Ethovision XT 7 pode controlar simultaneamente oito camundongos em tempo real). </li></ol></li><li> Procedimento de teste LMA longo prazo <ol><li> Teste LMA longo prazo durante a fase ativa / escuro do ciclo de luz. Certifique-se o tempo em relação ao ciclo de luz é a mesma em todos os grupos, como LMA segue um padrão crepuscular com mais atividade durante os períodos iniciais e depois do ciclo escuro 1. Teste os dois sujeitos experimentais e controle em cada grupo de camundongos gravadas para garantir a minimização do potencialefeitos das diferenças temporais na LMA. </li><li> Individualmente casa e aclimatar os ratos nas gaiolas de teste modificados, para, pelo menos, 24 horas. </li><li> Coloque as divisórias de gaiolas e jaulas (contendo camundongos individuais) diretamente sob a câmera. Remover todo o material de assentamento / enriquecimento das gaiolas para assegurar uma vista desimpedida do do rato. Rato enterrando-se o material de assentamento irá interferir com a capacidade do VTS para controlar o rato. Colocar uma tampa de Plexiglas claro em cada uma das gaiolas a ser gravados. </li><li> Ao gravar várias gaiolas de uma vez, ter certeza que a resolução é compatível com os VTS escolhidos. Para facilitar a análise VTS, posição idêntica gaiolas dentro de cada um divisor (ou seja, canto superior direito). </li><li> LMA testes a longo prazo é geralmente analisada em tempo real, e um registro de vídeo geralmente não é criado. A gravação de vídeo pode ser gravado usando uma câmera secundária, ou através de um gravador de DVD ligado à câmera principal. Certifique-se de manter registros detalhados de indivíduorato e informações sobre o grupo. </li><li> Configure os VTS para análise em tempo real. O chão da gaiola deve abranger todo o ROI para rastreamento. Configure ROIs para todas as gaiolas a ser gravado. Usar o VTS para calcular a distância total movida (cm), a velocidade de movimento (cm / seg) e da duração do movimento (seg). Na Figura 2, as variáveis ​​de detecção eram: 2 cm / seg começa rastreamento e 1,75 cm / s rastreamento pára. </li><li> Começar a gravar rato longo prazo LMA. Monitorar a duração do ensaio de LMA de longo prazo através do relógio associado com o computador analisar. </li><li> Durante o teste, manter todos os experimentadores fora da vista dos ratos de modo a não agir como um sinal visual externo. </li><li> Quando o teste estiver concluído, o retorno / substitua o material do assentamento / enriquecimento, para reduzir o potencial de desenvolvimento da ansiedade-como o comportamento 8. </li></ol></li><li> LMA análise de longo prazo <ol><li> Análise LMA longo prazo podem ocorrer em tempo real, sem a geração de uma gravação de vídeo. Exportar os dados para Microsoft Excel para armazenamento e análise, se estiver usando Ethovision XT 7. </li><li> Para gerar um registro de vídeo para LMA longo prazo, usar uma câmera secundária. Certifique-se a câmera secundária tem ampla capacidade de armazenamento suficiente para armazenar o potencialmente longa de gravação (s). </li></ol></li></ol>

<ol>
	<li>Goulding, E. H., Schenk, A. K., Juneja, P., MacKay, A. W., Wade, J. M., Tecott, L. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+robust+automated+system+elucidates+mouse+home+cage+behavioral+structure.">A robust automated system elucidates mouse home cage behavioral structure.</a> Proc. Natl. Acad. Sci. U.S.A. 105, 20575-20582 (2008).</li><li>Kent, S., Bluth&#233;, R. M., Kelley, K. W., Dantzer, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sickness+behavior+as+a+new+target+for+drug+development.">Sickness behavior as a new target for drug development.</a> Trends Pharmacol. Sci. 13, 24-28 (1992).</li><li>Dantzer, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cytokine,+sickness+behavior,+and+depression.">Cytokine, sickness behavior, and depression.</a> Immunol. Allergy Clin. North Am. 29, 247-264 (2009).</li><li>Godbout, J. P., 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=Exaggerated+neuroinflammation+and+sickness+behavior+in+aged+mice+following+activation+of+the+peripheral+innate+immune+system.">Exaggerated neuroinflammation and sickness behavior in aged mice following activation of the peripheral innate immune system.</a> FASEB J. 19, 1329-1331 (2005).</li><li>York, J. 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+biobehavioral+and+neuroimmune+impact+of+low-dose+ionizing+radiation.">The biobehavioral and neuroimmune impact of low-dose ionizing radiation.</a> Brain Behav. Immun. 26, 218-227 (2012).</li><li>York, J. 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=Individually+ventilated+cages+cause+chronic+low-grade+hypoxia+impacting+mice+hematologically+and+behaviorally.">Individually ventilated cages cause chronic low-grade hypoxia impacting mice hematologically and behaviorally.</a> Brain Behav. Immun. 26, 951-958 (2012).</li><li>Jennings, 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=Refining+rodent+husbandry:+the+mouse.+Report+of+the+Rodent+Refinement+Working+Party.">Refining rodent husbandry: the mouse. Report of the Rodent Refinement Working Party.</a> Lab. Anim. 32, 233-259 (1998).</li><li>York, J. M., Blevins, N. A., Baynard, T., Freund, G. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mouse+testing+methods+in+psychoneuroimmunology.">Mouse testing methods in psychoneuroimmunology.</a> Methods in Molecular Biology. 934, 243-276 (2012).</li><li>Dantzer, R., O'Connor, J. C., Freund, G. G., Johnson, R. W., Kelley, K. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=From+inflammation+to+sickness+and+depression:+when+the+immune+system+subjugates+the+brain.">From inflammation to sickness and depression: when the immune system subjugates the brain.</a> Nat. Rev. Neurosci. 9, 46-56 (2008).</li><li>Crawley, J. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Behavioral+phenotyping+of+rodents.">Behavioral phenotyping of rodents.</a> Comp. Med. 53, 140-146 (2003).</li><li>Karelina, K., 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=Social+isolation+alters+neuroinflammatory+response+to+stroke.">Social isolation alters neuroinflammatory response to stroke.</a> Proc. Natl. Acad. Sci. U.S.A. 106, 5895-5900 (2009).</li></ol>

Os autores declaram não haver conflitos de interesse.

Os sintomas da doença em camundongos incluem letargia, perda de interesse em ambientes sociais e ambientais, mal-estar e anorexia 2-3. Em ratos e outros roedores, LMA pode ser utilizado para avaliar a doença após a activação imunitária 4-5. Além disso, LMA longo prazo oferece a capacidade de acompanhar o movimento ao longo de 24 horas (ou mais) período, permitindo a análise de alterações induzidas pelo tratamento no comportamento circadiano ou distúrbios nos padrões de sono / vigília....

<table border="0" cellpadding="0" cellspacing="0" width="586">
	<colgroup>
		<col />
		<col />
		<col />
		<col />
	</colgroup>
	<tbody>
		<tr height="21">
			<td height="21">Name of Reagent/Material</td>
			<td>Company</td>
			<td>Catalog Number</td>
			<td>Comments</td>
		</tr>
		<tr height="276">
			<td height="276">Video camera</td>
			<td>Sony</td>
			<td>HDR-XR500V</td>
			<td>Other, similar video cameras may work as well. The file storage format (codec/container) must be compatible with the VTS used. It is recommended when initially establishing a VTS to choose the VTS first and work with the vendor in choosing an appropriate camera, especially with respects to file format and video resolution compatibility.</td>
		</tr>
		<tr height="331">
			<td height="331">Opaque cage dividers</td>
			<td>&nbsp;</td>
			<td>&nbsp;</td>
			<td>Custom fabricated based on specific cage dimensions. To facilitate image analysis, dividers should be of a similar color to the cage bedding. Structurally fixed dividers are preferrable because they speed setup between multi-mice experiments, such as those involving repeated measurements. Sturdy transportable cage dividers can be constructed of 1/2 inch opaque plastic and joined via a series of cross laps.</td>
		</tr>
		<tr height="74">
			<td height="74">Light or Dark bedding (opposite of mouse coat color)</td>
			<td>&nbsp;</td>
			<td>&nbsp;</td>
			<td>Available from several laboratory animal supply distributors, depending on location.</td>
		</tr>
		<tr height="37">
			<td height="37">Clear Plexiglas cage tops</td>
			<td>&nbsp;</td>
			<td>&nbsp;</td>
			<td>Custom fabricated based on cage dimensions.</td>
		</tr>
		<tr height="92">
			<td height="92">Automated video tracking software (EthoVision XT 7)</td>
			<td>Noldus Information Technology</td>
			<td>&nbsp;</td>
			<td>A variety of VTS manufacturers exist, use of other software may suffice.</td>
		</tr>
		<tr height="55">
			<td height="55">PC</td>
			<td>Dell Inc.</td>
			<td>&nbsp;</td>
			<td>See VTS software providers minimum PC .requirements</td>
		</tr>
		<tr height="83">
			<td height="83">White noise generator</td>
			<td>Dohm</td>
			<td>DS</td>
			<td>A variety of white noise generators are available, depending on need.</td>
		</tr>
		<tr height="146">
			<td height="146">Brooder clamp lights with red light bulbs</td>
			<td>&nbsp;</td>
			<td>&nbsp;</td>
			<td>Several sizes and styles available. Number required depends on room size/lighting needs. Do not exceed 125 lumens in the testing room.</td>
		</tr>
		<tr height="62">
			<td height="62">Lab bench or butcher paper</td>
			<td>&nbsp;</td>
			<td>&nbsp;</td>
			<td>Paper should match or be very similar in color to the cage bedding.</td>
		</tr>
	</tbody>
</table>

mouse short- and long-term locomotor activity analyzed by video tracking software

Atividade locomotora (LMA) é uma medida simples e de fácil execução de comportamento em ratos e outros roedores. Melhorias no software de monitoramento de vídeo (VTS) ter permitido que ele seja acoplado a LMA testes, melhorando drasticamente a sensibilidade e especificidade quando comparado com o método de cruzamentos de linha com a pontuação manual. Além disso, o VTS permite a experimentação de alto rendimento. Embora semelhante ao controle de vídeo automatizado usado para o teste de campo aberto (OFT), LMA teste é o único que permite a ratos para permanecer em sua gaiola e não utiliza o estímulo ansiogênico de iluminação brilhante durante a fase ativa do claro-escuro ciclo. Tradicionalmente, o LMA tem sido usada durante curtos períodos de tempo (minutos), enquanto os estudos de circulação mais longos (HRS-dia), muitas vezes têm utilizado transmissores e biotelemetria implantado. Com a opção de rastreamento em tempo real, de longo, como testes de LMA de curto prazo, podem agora ser realizados utilizando videografia. LMA testes a longo prazo requer um especialista, Mas facilmente construído, de modo que a gaiola de comida e água (o que é normalmente posicionado na parte superior da gaiola) não obstrua videografia. Importante, videografia e VTS permite a quantificação de parâmetros, como caminho de movimento de mouse, que são difíceis ou inviáveis ​​para medir com a linha de cruzamento e / ou biotelemetria. Em resumo, o teste LMA acoplado a VTS proporciona uma descrição mais completa do movimento do rato e a capacidade de examinar locomoção ao longo de um período de tempo prolongado.

Um exemplo dos resultados de uma série de medidas repetidas, teste de LMA de curto prazo é mostrado na Figura 1. Esta figura mostra que, após administração de lipopolissacarídeo (LPS), ratos alojados em enjaular ventilado individualmente (IVC) recuperar a atividade locomotora de curto prazo mais rápido do que os ratos alojados em ambiente ambiente prendendo (AEC) 6. Enquanto a perda inicial de actividade locomotora é semelhante entre os grupos injectados com LPS, a recuperação de ba...

Watch this Scientific Journal Video about Mouse Short- and Long-term Locomotor Activity Analyzed by Video Tracking Software at JoVE.com

Mouse Short- and Long-term Locomotor Activity Analyzed by Video Tracking Software

Atividade locomotora (LMA) é uma medida simples e de fácil execução de comportamento em camundongos. O acoplamento do software de rastreamento de vídeo (VTS) e LMA permite a melhoria da especificidade e sensibilidade, especialmente quando comparado com o manual, o método de análise cruzando a linha de LMA. Além disso VTS permite o acompanhamento a longo prazo de rato LMA.

Rato de curto e longo prazo da atividade locomotora analisados ​​pelo software Rastreamento Vídeo

Locomotor  activity (LMA) is a simple and easily performed measurement of behavior in mice  and other rodents. Improvements in video tracking software ...

mouse-short-long-term-locomotor-activity-analyzed-video-tracking

Research

JoVE Journal

Behavior

20.0K visualizações.  University of Illinois at Urbana-Champaign. Atividade locomotora (LMA) é uma medida simples e de fácil execução de comportamento em camundongos. O acoplamento do software de rastreamento de vídeo (VTS) e LMA permite a melhoria da especificidade e sensibilidade, especialmente quando comparado com o manual, o método de análise cruzando a linha de LMA. Além disso VTS permite o acompanhamento a longo prazo de rato LMA.

Vídeo: Mouse Short- and Long-term Locomotor Activity Analyzed by Video Tracking Software

Long-term Potentiation of Perforant Pathway-dentate Gyrus Synapse in Freely Behaving Mice

Studies of long-term potentiation of synaptic efficacy, an activity-dependent synaptic phenomenon having properties that make it attractive as a potential cellular mechanism underlying learning and information storage, have long been used to elucidate the physiology of various neuronal circuits in the hippocampus, amygdala, and other limbic and cortical structures. With this in mind, transgenic mouse models of neurological diseases represent useful platforms to conduct long-term&#160;potentiation (LTP) studies to develop a greater understanding of the role of genes in normal and abnormal synaptic communication in neuronal networks involved in learning, emotion and information processing. This article describes methodologies for reliably inducing LTP in the freely behaving mouse. These methodologies can be used in studies of transgenic and knockout freely&#160;behaving mouse models of neurodegenerative diseases.

Transgenic and knockout mouse models of neurological diseases are useful for studying the role of genes in normal and abnormal neurophysiology. This article describes methodologies which can be used to study long-term potentiation, a cellular mechanism which may underlie learning and memory, in transgenic and knockout freely&#160;behaving mouse models of neuropathology.

Potenciação de longo prazo da Via Perfurante-giro denteado Synapse em Livremente Comportando Ratos

Studies of long-term potentiation of synaptic efficacy, an activity-dependent synaptic phenomenon having properties that make it attractive as a potential ...

Comportamento

Behavioral and Locomotor Measurements Using an Open Field Activity Monitoring System for Skeletal Muscle Diseases

The open field activity monitoring system comprehensively assesses locomotor and behavioral activity levels of mice. It is a useful tool for assessing locomotive impairment in animal models of neuromuscular disease and efficacy of therapeutic drugs that may improve locomotion and/or muscle function. The open field activity measurement provides a different measure than muscle strength, which is commonly assessed by grip strength measurements. It can also show how drugs may affect other body systems as well when used with additional outcome measures. In addition, measures such as total distance traveled mirror the 6 min walk test, a clinical trial outcome measure. However, open field activity monitoring is also associated with significant challenges: Open field activity measurements vary according to animal strain, age, sex, and circadian rhythm. In addition, room temperature, humidity, lighting, noise, and even odor can affect assessment outcomes. Overall, this manuscript provides a well-tested and standardized open field activity SOP for preclinical trials in animal models of neuromuscular diseases. We provide a discussion of important considerations, typical results, data analysis, and detail the strengths and weaknesses of open field testing. In addition, we provide recommendations for optimal study design when using open field activity in a preclinical trial.

Open field activity levels are used to assess locomotive and behavioral activity levels. This protocol provides a well-designed, standardized protocol to use in preclinical trials for neuromuscular disorders.

Medidas comportamentais e do aparelho locomotor Usando um sistema aberto Activity Monitoring Campo de Doenças músculo-esquelético

The open field activity monitoring system comprehensively assesses locomotor and behavioral activity levels of mice. It is a useful tool for assessing ...

Using a Split-belt Treadmill to Evaluate Generalization of Human Locomotor Adaptation

Understanding the mechanisms underlying locomotor learning helps researchers and clinicians optimize gait retraining as part of motor rehabilitation. However, studying human locomotor learning can be challenging. During infancy and childhood, the neuromuscular system is quite immature, and it is unlikely that locomotor learning during early stages of development is governed by the same mechanisms as in adulthood. By the time humans reach maturity, they are so proficient at walking that it is difficult to come up with a sufficiently novel task to study de novo locomotor learning. The split-belt treadmill, which has two belts that can drive each leg at a different speed, enables the study of both short- (i.e., immediate) and long-term (i.e., over minutes-days; a form of motor learning) gait modifications in response to a novel change in the walking environment. Individuals can easily be screened for previous exposure to the split-belt treadmill, thus ensuring that all experimental participants have no (or equivalent) prior experience. This paper describes a typical split-belt treadmill adaptation protocol that incorporates testing methods to quantify locomotor learning and generalization of this learning to other walking contexts. A discussion of important considerations for designing split-belt treadmill experiments follows, including factors like treadmill belt speeds, rest breaks, and distractors. Additionally, potential but understudied confounding variables (e.g., arm movements, prior experience) are considered in the discussion.

We describe a protocol for investigating human locomotor adaptation using the split-belt treadmill, which has two belts that can drive each leg at a different speed. We specifically focus on a paradigm designed to test the generalization of adapted locomotor patterns to different walking contexts (e.g., gait speeds, walking environments).

Usando uma escada rolante Split-cinto para avaliar a generalização de adaptação de locomoção humana

Understanding the mechanisms underlying locomotor learning helps researchers and clinicians optimize gait retraining as part of motor rehabilitation. ...

A Novel Single Animal Motor Function Tracking System Using Simple, Readily Available Software

We have recently demonstrated that implanting intracortical microelectrodes in the motor corteces of rats&#160;results in immediate and lasting motor deficits. Motor impairments were manually quantified through an open field grid test to measure the gross motor function and through a ladder test to measure the fine motor function. Here, we discuss a technique for the automated quantification of the video-recorded tests using our custom Capadona Behavioral Video Analysis System: Grid and Ladder Test, or BVAS. Leveraging simple and readily available coding software (see the Table of Materials), this program allows for the tracking of a single animal on both the open field grid and the ladder tests. In open field grid tracking, the code thresholds the video for intensity, tracks the position of the rat over the 3 min duration of the grid test, and analyzes the path. It then computes and returns measurements for the total distance traveled, the maximum velocity achieved, the number of left- and right-handed turns, and the total number of grid lines crossed by the rat. In ladder tracking, the code again thresholds the video for intensity, tracks the movement of the rat across the ladder, and returns calculated measurements including the time it took the rat to cross the ladder, the number of paw slips occurring below the plane of the ladder rungs, and the incidence of failures due to stagnation or reversals. We envision that the BVAS developed here can be employed for the analysis of motor function in a variety of applications, including many injury or disease models.

The current study aimed to automate the quantification of motor deficits in rats. The initial evaluation model assesses motor loss resulting from an intracortical microelectrode implantation in the motor cortex. We report on the development and use of a tracking algorithm using easily adaptable, simple, and readily available coding software.

Uma romance única Animal motricidade sistema usando simples, prontamente disponível Software de rastreamento

We have recently demonstrated that implanting intracortical microelectrodes in the motor corteces of rats&#160;results in immediate and lasting motor ...

Automated, Long-term Behavioral Assay for Cognitive Functions in Multiple Genetic Models of Alzheimer's Disease, Using IntelliCage

Multiple factors&#8212;such as aging and genes&#8212;are frequently associated with cognitive decline. Genetically modified mouse models of cognitive decline, such as Alzheimer&#39;s disease (AD), have become a promising tool to elucidate the underlying mechanisms and promote the therapeutic advances. An important step is the validation and characterization of expected behavioral abnormality in the models, in the case of AD, cognitive decline. The long-term behavioral investigations of laboratory animals to study the effect of aging demand substantial efforts from researchers. The IntelliCage system is a high-throughput and cost-effective test battery for mice that eliminates the need for daily human handling. Here, we describe how the system is utilized in the long-term phenotyping of a genetic Alzheimer&#39;s disease model, specifically focusing on the cognitive functions. The experiment employs repeated battery of tests that assess spatial learning and executive functions. This cost-effective age-dependent phenotyping allows us to identify the transient and/or permanent effects of genes on various cognitive aspects.

Automated, Long-term Behavioral Assay for Cognitive Functions in Multiple Genetic Models of Alzheimer&#039;s Disease, Using IntelliCage

This paper describes a protocol for cognitive assessments for genetic models of the Alzheimer&#39;s disease using the IntelliCage system, which is a high throughput automated behavioral monitoring system with operant conditioning.

Automatizada, a longo prazo ensaio comportamental para funções cognitivas em vários modelos genéticos da doença de Alzheimer, usando IntelliCage

Multiple factors&#8212;such as aging and genes&#8212;are frequently associated with cognitive decline. Genetically modified mouse models of cognitive ...

Classical Short-Delay Eyeblink Conditioning in One-Year-Old Children

Classical eyeblink conditioning (EBC) refers to the learned association between a conditioned stimulus (an auditory tone) and an unconditioned stimulus (a puff of air to the cornea). Eyeblink conditioning is often used experimentally to detect abnormalities in cerebellar-dependent learning and memory that underlies this type of associative learning. While experiments in adults and older children are relatively simple to administer using commercial equipment, eyeblink conditioning in infants is more challenging due to their poor compliance, which makes correct positioning of the equipment difficult. To achieve conditioning in one-year-old infants, a custom-made or an adapted commercial system can be used to deliver the air puff to the infant's cornea. The main challenge lies in successfully detecting and classifying the behavioral responses. We report that automated blink detection methods are unreliable in this population, and that conditioning experiments should be analyzed using frame-by-frame analysis of supplementary video camera recordings. This method can be applied to study developmental changes in eyeblink conditioning and to examine whether this paradigm can detect children with neurological disorders.

This protocol describes an eyeblink conditioning paradigm appropriate for experiments with one-year-old infants. Commercial or custom-made equipment can be used to deliver the stimuli, and data collection and analysis should be performed on the video recordings.

Condicionamento clássico curto-atraso Eyeblink em crianças de um ano de idade

Classical eyeblink conditioning (EBC) refers to the learned association between a conditioned stimulus (an auditory tone) and an unconditioned stimulus (a ...

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 ...

Elevated Plus Maze Test Combined with Video Tracking Software to Investigate the Anxiolytic Effect of Exogenous Ketogenic Supplements

The overall goal of this study is to describe the methodology of the elevated plus maze (EPM) test in combination with a video tracking software. The purpose of the method is to document the effect of various potential anxiolytic treatments on laboratory rodent models. The EPM test is based on the rodents&#39; proclivity toward protected, enclosed dark spaces and unconditioned fear of open spaces and heights, and their innate intense motivation to explore novel environments. The EPM test is a widely used behavioral test for investigating the anxiolytic or anxiogenic responses of rodents given drugs that are known to affect behavior. Observation demonstrating a decreased proportion of time spent on closed arms, an increased proportion of time spent on open arms, a reduced number of entries to closed arms, and an elevated number of entries to open arms measured by the EPM test may reflect reduced anxiety levels. Using this method, the effect of exogenous ketone supplements on anxiety-related behavior is tested in Sprague Dawley (SPD) rats. Exogenous ketone supplements are chronically fed to the rats for 83 days or subchronically and acutely orally gavaged, daily for 7 days, before conducting the EPM test. Behavioral data collection is performed using the SMART video tracking system by a blinded observer at the end of the treatments. The main findings indicate that the EPM test is an effective method to detect the ketone supplement-induced anxiolytic effect and can be considered a sensitive measure to assess changes in anxiety behavior associated with drug- or metabolic-based therapies.

Here, we present a protocol to investigate changes in the anxiety level of rodent animal models. The elevated plus maze (EPM) test, used together with a video tracking software, provides a reliable method to document the effect of various potential anxiolytic treatments in preclinical laboratory scenarios.

Elevado, mais teste combinado com vídeo que segue o Software para investigar o efeito ansiolítico dos suplementos cetogênica exógenos de labirinto

The overall goal of this study is to describe the methodology of the elevated plus maze (EPM) test in combination with a video tracking software. The ...

Behavioral Tracking and Neuromast Imaging of Mexican Cavefish

Cave-dwelling animals have evolved a series of morphological and behavioral traits to adapt to their perpetually dark and food-sparse environments. Among these traits, foraging behavior is one of the useful windows into functional advantages of behavioral trait evolution. Presented herein are updated methods for analyzing vibration attraction behavior (VAB: an adaptive foraging behavior) and imaging of associated mechanosensors of cave-adapted tetra, Astyanax mexicanus. In addition, methods are presented for high-throughput tracking of a series of additional cavefish behaviors including hyperactivity and sleep-loss. Cavefish also show asociality, repetitive behavior and higher anxiety. Therefore, cavefish serve as an animal model for evolved behaviors. These methods use free-software and custom-made scripts that can be applied to other types of behavior. These methods provide practical and cost-effective alternatives to commercially available tracking software.

Here, we present methods for high-throughput study of a series of the Mexican cavefish behaviors and vital staining of a mechanosensory system. These methods use free-software and custom-made scripts, providing a practical and cost-effective method for the studies of behaviors.

Acompanhamento comportamental e Neuromast imagem de Cavefish mexicano

Cave-dwelling animals have evolved a series of morphological and behavioral traits to adapt to their perpetually dark and food-sparse environments. Among ...

Automated Measurements of Sleep and Locomotor Activity in Mexican Cavefish

Across phyla, sleep is characterized by highly conserved behavioral characteristics that include elevated arousal threshold, rebound following sleep deprivation, and consolidated periods of behavioral immobility. The Mexican cavefish, Astyanax mexicanus (A. mexicanus), is a model for studying trait evolution in response to environmental perturbation. A. mexicanus exist as in eyed surface-dwelling forms and multiple blind cave-dwelling populations that have robust morphological and behavioral differences. Sleep loss has occurred in multiple, independently-evolved cavefish populations. This protocol describes a methodology for quantifying sleep and locomotor activity in A. mexicanus cave and surface fish. A cost-effective video monitoring system allows for behavioral imaging of individually-housed larval or adult fish for periods of a week or longer. The system can be applied to fish aged 4 days post fertilization through adulthood. The approach can also be adapted for measuring the effects of social interactions on sleep by recording multiple fish in a single arena. Following behavioral recordings, data is analyzed using automated tracking software and sleep analysis is processed using customized scripts that quantify multiple sleep variables including duration, bout length, and bout number. This system can be applied to measure sleep, circadian behavior, and locomotor activity in almost any fish species including zebrafish and sticklebacks.

This protocol details the methodology for quantifying locomotor behavior and sleep in the Mexican cavefish. Previous analyses are extended to measure these behaviors in socially-housed fish. This system can be widely applied to study sleep and activity in other fish species.

Medições automatizadas do sono e da atividade locomotora em Cavefish mexicano

Across phyla, sleep is characterized by highly conserved behavioral characteristics that include elevated arousal threshold, rebound following sleep ...

Rato de curto e longo prazo da atividade locomotora analisados ​​pelo software Rastreamento Vídeo

Rato de curto e longo prazo da atividade locomotora analisados pelo software Rastreamento Vídeo