Name: chronic sleep deprivation in mouse pups by means of gentle handling
Uploaded: 2018-10-11T13:30:00
Description: Watch this Scientific Journal Video about Chronic Sleep Deprivation in Mouse Pups by Means of Gentle Handling at JoVE.com

Esta pesquisa foi financiada pelo programa de pesquisa Intramural do NIMH (ZIA MH00889). RMS também foi apoiado por uma bolsa de pós-doutorado FRAXA.

Nota: Todos os procedimentos foram aprovados pelo Comitê de uso e National Institute de Mental Saúde Cuidado Animal e efectuados de acordo com os institutos nacionais de saúde orientações sobre os cuidados e uso de animais.
1. instalação de privação de sono
<ol><li>Use pincéis macios para picar delicadamente os ratos quando dormindo.<ol>
<li>Certifique-se que o pincel pode caber através das grades do alimentador gaiola do rato para garantir que os ratos têm acesso à comida e ág...

<ol>
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L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sleep+and+pain:+A+relationship+that+begins+in+early+life.">Sleep and pain: A relationship that begins in early life.</a> Pain Physician. 17 (6), E787-E798 (2014).</li><li>Hairston, I. S., 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=Sleep+deprivation+effects+on+growth+factor+expression+in+neonatal+rats:+a+potential+role+for+BDNF+in+the+mediation+of+delta+power.">Sleep deprivation effects on growth factor expression in neonatal rats: a potential role for BDNF in the mediation of delta power.</a> J Neurophysiol. 91 (4), 1586-1595 (2004).</li><li>Hairston, I. S., 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=Sleep+deprivation+elevates+plasma+corticosterone+levels+in+neonatal+rats.">Sleep deprivation elevates plasma corticosterone levels in neonatal rats.</a> Neurosci Lett. 315 (1-2), 29-32 (2001).</li><li>Blumberg, M. S., Middlemis-Brown, J. E., Johnson, E. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sleep+homeostasis+in+infant+rats.">Sleep homeostasis in infant rats.</a> Behav Neurosci. 118 (6), 1253-1261 (2004).</li><li>Feng, P., Vogel, G. W., Obermeyer, W., Kinney, G. 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B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Circadian+rhythm+of+melatonin,+corticosterone+and+phagocytosis:+Effect+of+stress.">Circadian rhythm of melatonin, corticosterone and phagocytosis: Effect of stress.</a> J Pineal Res. 30 (3), 180-187 (2001).</li><li>Nadler, J. J., 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=Automated+apparatus+for+quantitation+of+social+approach+behaviors+in+mice.">Automated apparatus for quantitation of social approach behaviors in mice.</a> Genes Brain Behav. 3 (5), 303-314 (2004).</li><li>Thomas, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Marble+burying+reflects+a+repetitive+and+perseverative+behavior+more+than+novelty-induced+anxiety.">Marble burying reflects a repetitive and perseverative behavior more than novelty-induced anxiety.</a> Psychopharmacology (Berl). 204 (2), 361-373 (2009).</li><li>Karlsson, K. A., Blumberg, M. 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Aqui nós descrevemos um método para a privação de sono, suave manipulação, que pode ser usada em ambos os filhotes de roedores e em adultos. Entrega de suave requer os pesquisadores a observar os ratos para a duração de um período de privação de sono e suavemente prod os animais quando eles estão inativos ou contrair para impedir o sono. Estudos de privação de sono prévio verificou-se por meio de gravações simultâneas de eletroencefalograma (EEG), que suave manipulação impede que ambos REM e NREM não...

Sono desempenha um papel fundamental na formação de sinapse e plasticidade neuronal e eliminação durante o desenvolvimento de cérebro1,2. Especificamente, o sono de movimento rápido dos olhos (REM) é essencial para a formação de circuitos sinápticos estáveis através de ambos fortalecimento das sinapses específicas e poda sináptica2. Privação de sono farmacológicos no início da vida leva a muitos déficits fisiológicos e comportamentais3,4. Além de farmacológico sono privação, outras formas de privação do sono, tais como vibração constante, forçado a locomoção, ou apresentação de um estímulo contrário têm sido associados com sintomas de depressão durante a vida adulta, diferente neural padrões de ativação e mudanças na continuidade de tempo e sono sono durante um período de recuperação após privação5,6,7. Quando o sono é cronicamente restrito em camundongos há mais de cinco semanas, durante o desenvolvimento, os défices comportamentais foram observados após um mês de sono recuperação8. Tomados em conjunto, estes estudos sugerem que a perturbação do sono no roedor neonatal pode afetar posteriores padrões de sono, a função cerebral e comportamento. Destacando a importância do sono para o desenvolvimento normal do cérebro em humanos, muitos pacientes com transtornos do desenvolvimento neurológico, incluindo transtornos do espectro do autismo, complexo de esclerose tuberosa, síndrome de X frágil e atenção-déficit Desordem, anormalidades do relatório nos padrões de sono, além de uma variedade de défices comportamentais1.
Dado o número de transtornos do desenvolvimento neurológico que estão associados com padrões de sono anormais, é importante entender como falta de sono afeta o comportamento e a função cerebral. No entanto, apesar dos dados que suportam a importância do sono durante o desenvolvimento, a maioria das metodologias que restringem ou privam os animais de foco do sono em adultos. Estas técnicas incluem uma variedade de testes que exigem forçado a atividade locomotora (por exemplo, constante da esteira), perturbação constante (por exemplo, rotação bar vassoura ou apresentação contínua objeto romance) ou perturbar o animal no início da 9,de sono REM (por exemplo, a plataforma sobre a água)10. Embora esses métodos demonstraram efetivamente os problemas associados com a privação do sono em adultos, nenhum desses métodos são apropriados para filhotes de roedores devido sua mobilidade limitada, unidade de sono alta e os efeitos adversos do stress no início da vida. Devido à importância do sono durante o desenvolvimento, é fundamental para entender o sono como restrito no comportamento futuro do afeta neonatal filhotes durante a vida adulta.
Uma técnica para restringir o sono que pode ser usado em filhotes de roedores é manipulação suave. Manipulação suave envolve o investigador interagindo diretamente com os roedores que qualquer hora sono comportamento é observado. Os investigadores podem usar suas mãos, um pincel ou apresentação de novos objetos fisicamente perturbar o sono em roedores neonatal8,10,11,12,13,14 . Sono pode ser determinado de forma comportamental por falta de atividade motora, espasmos Mioclônicas e fechamento do olho (em filhotes de roedores mais velhos). Validação de EEG confirma que interromper o sono com a manipulação suave baseada esses comportamentos reduz o tempo total de sono em 91% no P12 ratos14. Outras técnicas que foram usadas para privar os ratos jovens incluem choque elétrico e apresentação de estímulos desagradáveis, mas essas técnicas não podem ser usadas para restrição de sono crônica e são mais estressantes do que manipulação suave7, 15. restrição de sono crônica pode também ser realizada através de um protocolo de agitação que pode ser usado com ou sem implante de eletrodos de EEG e software de computador associado, mas este protocolo não impede o sono de todos e, portanto, é menos controlada em comparação com manipulação suave4,16. A capacidade de restringir ou privar ratos de sono por muitos dias seguidos sem equipamentos específicos, software de computador ou implantação do eletrodo é um benefício a técnica de manipulação suave. Restrição de sono crônica através de manipulação suave efetivamente limita o sono em roedores Neonatais e produz uma variedade de mudanças de comportamento após o sono privação período8,11,12, 17.
Aqui, descrevemos os resultados em que privação de sono crônica por três horas por dia pós-Natal dia 5 (P5)-P42 afeta significativamente sociabilidade de ratos após um período de recuperação de 30 dias em que o sono não seja perturbado. Esses resultados destacam os efeitos a longo prazo de restrição de sono crônica durante o desenvolvimento no comportamento futuro.

<table>
	<tbody>
		<tr>
			<td>C57BL/6J mice</td>
			<td>Jackson Labs</td>
			<td>000664</td>
			<td>Wildtype mice used</td>
		</tr>
		<tr>
			<td>Super Mouse 750 Mouse Cage</td>
			<td>Lab Products, Inc.&#160;</td>
			<td></td>
			<td>Cages for the mice</td>
		</tr>
		<tr>
			<td>SANI-Chips Bedding</td>
			<td>PJ Murphys</td>
			<td></td>
			<td>Bedding for the mice</td>
		</tr>
		<tr>
			<td>S&#38;S Bristle Brush Assortment Pack</td>
			<td>Staples</td>
			<td>Item # 13764, Model AB37100</td>
			<td>Paint brushes are used to gently poke mice</td>
		</tr>
	</tbody>
</table>

chronic sleep deprivation in mouse pups by means of gentle handling

O sono é fundamental para o bom desenvolvimento e plasticidade neural. Além disso, padrões de sono anormais são características de muitos transtornos do desenvolvimento neurológico. Restrição de sono estudando como crônica durante o desenvolvimento pode afetar o comportamento adulto pode aumentar a nossa compreensão do surgimento dos sintomas comportamentais de transtornos do desenvolvimento neurológico. Embora existam muitos métodos que podem ser usados para restringir o sono em roedores, incluindo locomoção forçada, perturbação constante, apresentação de um estímulo contrário ou choque elétrico, muitos destes métodos são muito estressantes e não podem ser usados em camundongos neonatais. Aqui, descrevemos manipulação suave, uma técnica de privação de sono que pode ser usada cronicamente durante todo o desenvolvimento e na idade adulta para alcançar a restrição de sono. Manipulação suave envolve estreita observação dos ratos durante todo o período de privação de sono e requer o pesquisador prod suavemente os animais quando eles estão inativos ou exibir comportamentos associados com o sono. Juntamente com as gravações de EEG, manipulação suave poderia ser usada para interromper seletivamente uma fase específica do sono, como sono de movimento rápido dos olhos (REM). A técnica de manipulação suave é uma poderosa ferramenta para o estudo dos efeitos da restrição de sono crônica mesmo em ratos neonatais que contorna a muitos dos procedimentos utilizados para a privação do sono mais estressantes.

Para investigar os efeitos da restrição de sono crônica sobre comportamento, dormimos ratos restritos por três horas diariamente entre as 11:00 e 14:00 de P5 P428. Após a restrição de sono, ratos foram deixados inalterados durante quatro semanas e permitiu recuperar da restrição de sono. Os ratos foram testados pelo comportamento após o período de quatro semanas de recuperação. Foi utilizado o teste de comportamento social três-câmara para avaliar a ...

Watch this Scientific Journal Video about Chronic Sleep Deprivation in Mouse Pups by Means of Gentle Handling at JoVE.com

Chronic Sleep Deprivation in Mouse Pups by Means of Gentle Handling

Nós descrevemos uma técnica de privação de sono conhecida como manipulação suave, onde os investigadores suavemente prod ratos qualquer momento sono comportamento é observado. Este método é uma ferramenta poderosa que permite aos pesquisadores estudar os efeitos que a restrição de sono crônica durante todo o desenvolvimento pode ter no comportamento e fisiologia do cérebro futuras.

Privação de sono crônica no Mouse Puppies suave através de movimentação

Sleep is critical for proper development and neural plasticity. Moreover, abnormal sleep patterns are characteristic of many neurodevelopmental disorders. ...

Este método pode ajudar a responder a perguntas-chave no campo do sono, como os efeitos da restrição crônica do sono durante o desenvolvimento pós-natal. A principal vantagem dessa técnica é que ela é menos estressante do que outras técnicas de privação do sono e pode ser facilmente usada durante todo o desenvolvimento e continuar até a idade adulta. Comece atribuindo cada gaiola à privação do sono ou ao grupo de controle.Certifique-se de que os ratos tenham acesso a alimentos e anúncios de água libitum, e remova qualquer alimento, roupa de cama ou objetos de enriquecimento que possam restringir a visão dos ratos. Para o grupo controle, retire o pai da gaiola do criador no quinto dia pós-natal e deixe a represa com os filhotes até desmamar. Em seguida, mova os ratos atribuídos ao grupo controle para a sala de privação do sono e gentilmente impulsione os ratos com um pincel continuamente por 10 minutos por dia durante a duração do experimento.Após 10 minutos, devolva os ratos para a sala de espera dos animais e não os perturbe. Comece removendo o pai da gaiola do criador no quinto dia pós-natal e deixe a represa com os filhotes até desmamar. Mova os ratos atribuídos ao grupo de privação do sono para a sala de privação do sono diariamente durante o ciclo de luz onde serão monitorados.Durante este tempo, gentilmente prod o mouse com um pincel se suspeitar de estar dormindo até que uma resposta é observada. Alternativamente, inverta ratos e empurre para suas costas para interromper o sono. Considere um rato adormecer se ocorrer algum dos seguintes: contração, inatividade ou se os olhos estiverem fechados.Considere os ratos acordados se eles estão se movendo, tentando virar depois de estar em suas costas, ou se preparando. Se a represa está cobrindo os filhotes durante a privação do sono, gentilmente afaste-a dos filhotes para que a visão dos filhotes seja irrestrita. Continue com o protocolo de privação do sono até o ponto final do experimento.Quando o período de três horas de privação do sono estiver completo, coloque a cama e qualquer alimento que possa ter sido removido de volta e coloque a tampa de volta na gaiola. Finalmente, devolva as gaiolas para a sala de espera dos animais até o dia seguinte da privação do sono. Após um período de recuperação de um mês, os resultados indicaram que, no teste de sociabilidade, as fêmeas restritas ao sono passaram mais tempo farejando o rato estranho do que controlando as fêmeas e os machos restritos ao sono tinham a tendência de passar mais tempo farejando o objeto quando comparados ao controle masculino.Além disso, na fase de novidade social dos testes, camundongos fêmeas restritas ao sono passaram um tempo maior farejando o rato novo do que os controles femininos. Ao tentar esse procedimento, é importante lembrar de prestar atenção e responder prontamente aos comportamentos pré-sono, especialmente em camundongos neonatais onde a necessidade de sono é alta. Após esse procedimento, outros métodos como campo aberto, rotarod, labirinto elevado e teste de comportamento social podem ser realizados a fim de responder a perguntas adicionais como os efeitos comportamentais de longo prazo da restrição crônica do sono durante o desenvolvimento.

chronic-sleep-deprivation-in-mouse-pups-by-means-of-gentle-handling

Research

JoVE Journal

Behavior

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

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 ...

Errors as a Means of Reducing Impulsive Food Choice

Nowadays, the increasing incidence of eating disorders due to poor self-control has given rise to increased obesity and other chronic weight problems, and ultimately, to reduced life expectancy. The capacity to refrain from automatic responses is usually high in situations in which making errors is highly likely. The protocol described here aims at reducing imprudent preference in women during hypothetical intertemporal choices about appetitive food by associating it with errors. First, participants undergo an error task where two different edible stimuli are associated with two different error likelihoods (high and low). Second, they make intertemporal choices about the two edible stimuli, separately. As a result, this method decreases the discount rate for future amounts of the edible reward that cued higher error likelihood, selectively. This effect is under the influence of the self-reported hunger level. The present protocol demonstrates that errors, well known as motivationally salient events, can induce the recruitment of cognitive control, thus being ultimately useful in reducing impatient choices for edible commodities.

Giving in to temptation of tasty food may result in long-term overweight problems. This protocol describes how to reduce imprudent preference for edible commodities during hypothetical intertemporal choices in women by associating them with errors.

Erros como um meio de reduzir Impulsivo Food escolha

Nowadays, the increasing incidence of eating disorders due to poor self-control has given rise to increased obesity and other chronic weight problems, and ...

Manipulation of Epileptiform Electrocorticograms (ECoGs) and Sleep in Rats and Mice by Acupuncture

Ancient Chinese literature has documented that acupuncture possesses efficient therapeutic effects on epilepsy and insomnia. There is, however, little research to reveal the possible mechanisms behind these effects. To investigate the effect of acupuncture on epilepsy and sleep, several issues need to be addressed. The first is to identify the acupoints, which correspond between humans, rats, and mice. Furthermore, the depth of insertion of the acupuncture needle, the degree of needle twist in manual needle acupuncture, and the stimulation parameters for electroacupuncture (EA) need to be determined. To evaluate the effects of acupuncture on epilepsy and sleep, a feasible model of epilepsy in rodents is required. We administer pilocarpine into the left central nucleus of the amygdala (CeA) to simulate focal temporal lobe epilepsy (TLE) in rats. Intraperitoneal (IP) injection of pilocarpine induces generalized epilepsy and status epilepticus (SE) in rats. Five IP injections of pentylenetetrazol (PTZ) with a one-day interval between each injection successfully induces spontaneous generalized epilepsy in mice. Recordings of electrocorticograms (ECoGs), electromyograms (EMGs), brain temperature, and locomotor activity are used for sleep analysis in rats, while ECoGs, EMGs, and locomotor activity are employed for sleep analysis in mice. ECoG electrodes are implanted into the frontal, parietal, and contralateral occipital cortices, and a thermistor is implanted above the cerebral cortex by stereotactic surgery. EMG electrodes are implanted into the neck muscles, and an infrared detector determines locomotor activity. The criteria for categorizing vigilance stages, including wakefulness, rapid eye movement (REM) sleep, and non-REM (NREM) sleep are based on information from ECoGs, EMGs, brain temperature, and locomotor activity. Detailed classification criteria are stated in the text.

Manipulation of Epileptiform Electrocorticograms ECoGs and Sleep in Rats and Mice by Acupuncture

This paper demonstrates the performance of acupuncture, epilepsy models, and the analysis of sleep in rodents. The acupuncture procedure and the identification of acupoints are described. Pilocarpine or pentylenetetrazol (PTZ) is used to induce epilepsy. Electrocorticogram (ECoG), electromyogram (EMG), brain temperature, and locomotor activity recordings are employed for sleep analysis.

Manipulação de epileptiforme Electrocorticograms (ECoGs) e do sono em ratos e camundongos por Acupuntura

Ancient Chinese literature has documented that acupuncture possesses efficient therapeutic effects on epilepsy and insomnia. There is, however, little ...

Neurodevelopmental Reflex Testing in Neonatal Rat Pups

Neurodevelopmental reflex testing is commonly used in clinical practice to assess the maturation of the nervous system. Neurodevelopmental reflexes are also referred to as primitive reflexes. They are sensitive and consistent with later outcomes. Abnormal reflexes are described as an absence, persistence, reappearance, or latency of reflexes, which are predictive indices of infants that are at high risk for neurodevelopmental disorders. Animal models of neurodevelopmental disabilities, such as cerebral palsy, often display aberrant developmental reflexes, as would be observed in human infants. The techniques described assess a variety of neurodevelopmental reflexes in neonatal rats. Neurodevelopmental reflex testing offers the investigator a testing method that is not otherwise available in such young animals. The methodology presented here aims to assist investigators in examining developmental milestones in neonatal rats as a method of detecting early-onset brain injury and/or determining the effectiveness of therapeutic interventions. The methodology presented here aims to provide a general guideline for investigators.

Behavioral testing is the gold standard for determining outcomes following brain injury, and can identify the presence of developmental disabilities in infants and children. Neurodevelopmental reflexes are an early indicator of these abnormalities. A host of easily accomplished developmental reflex tests in the neonatal rodent were developed and described here.

Desenvolvimento neurológico Reflex Teste em Neonatais rato filhotes de cachorro

Neurodevelopmental reflex testing is commonly used in clinical practice to assess the maturation of the nervous system. Neurodevelopmental reflexes are ...

Noninvasive, High-throughput Determination of Sleep Duration in Rodents

Traditionally, sleep is monitored by an electroencephalogram (EEG). EEG studies in rodents require surgical implantation of the electrodes followed by a long recovery period. To perform an EEG recording, the animal is connected to a receiver, creating an unnatural tether to the head-mount. EEG monitoring is time consuming, carries risk to the animal, and is not a completely natural setting for the measurement of sleep. Alternative methods to detect sleep, particularly in a high-throughput fashion, would greatly advance the field of sleep research. Here, we describe a validated method for detecting sleep via activity-based home-cage monitoring. Previous studies have shown that sleep assessed via this method has a high degree of agreement with sleep defined by traditional EEG-based measures. Whereas this method is validated for total sleep time, it is important to note that sleep bout duration should be assessed by an EEG which has better temporal resolution. The EEG can also differentiate rapid eye movement (REM) and non-REM sleep, giving more detail about the exact nature of sleep. Nevertheless, activity-based sleep determination can be used to analyze multiple days of undisturbed sleep and to assess sleep as a response to an acute event (like stress). Here, we show the power of this system to detect the response of mice to daily intraperitoneal injections.

We describe a high-throughput method of measuring sleep by means of activity-based home-cage monitoring. This method offers advantages over traditional EEG-based methods. It is well validated for the determination of total sleep duration and can be a powerful tool to monitor sleep in rodent models of human disease.

Determinação não-invasiva, elevado-throughput da duração do sono em roedores

Traditionally, sleep is monitored by an electroencephalogram (EEG). EEG studies in rodents require surgical implantation of the electrodes followed by 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 ...

Novel Object Recognition and Object Location Behavioral Testing in Mice on a Budget

Ethologically relevant behavioral testing is a critical component of any study that uses mouse models to study the cognitive effects of various physiological or pathological changes. The object location task (OLT) and the novel object recognition task (NORT) are two effective behavioral tasks commonly used to reveal the function and relative health of specific brain regions involved in memory. While both of these tests exploit the inherent preference of mice for the novelty to reveal memory for previously encountered objects, the OLT primarily evaluates spatial learning, which relies heavily on hippocampal activity. The NORT, in contrast, evaluates non-spatial learning of object identity, which relies on multiple brain regions. Both tasks require an open-field-testing arena, objects with equivalent intrinsic value to mice, appropriate environmental cues, and video recording equipment and the software. Commercially available systems, while convenient, can be costly. This manuscript details a simple, cost-effective method for building the arenas and setting up the equipment necessary to perform the OLT and NORT. Furthermore, the manuscript describes an efficient testing protocol that incorporates both OLT and NORT and provides typical methods for data acquisition and analysis, as well as representative results. Successful completion of these tests can provide valuable insight into the memory function of various mouse model systems and appraise the underlying neural regions that support these functions.

Here we provide a protocol which includes comprehensive instructions for the economical establishment of murine object location and novel object recognition behavioral testing, including the design, cost, and construction of required equipment as well as execution of behavioral testing, data collection, and analysis.

Reconhecimento de objetos novos e localização de objeto comportamental testes em ratos em um orçamento

Ethologically relevant behavioral testing is a critical component of any study that uses mouse models to study the cognitive effects of various ...

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 ...

A Chronic Sleep Fragmentation Model using Vibrating Orbital Rotor to Induce Cognitive Deficit and Anxiety-Like Behavior in Young Wild-Type Mice

Sleep disturbance is generally common in populations as a chronic disease or a complained event. Chronic sleep disturbance is proposed to be closely linked to the pathogenesis of diseases, especially neurodegenerative diseases. We recently found that 2 months of sleep fragmentation initiated Alzheimer&#8217;s disease (AD)-like behavioral and pathological changes in young wild-type mice. Herein, we present a standardized protocol to achieve chronic sleep fragmentation (CSF). Briefly, CSF was induced by an orbital rotor vibrating at 110 rpm and operating with a repetitive cycle of 10 s-on, 110 s-off, during light-ON phase (8:00 AM&#8211;8:00 PM) continuously for up to 2 months. Impairments of spatial learning and memory, anxiety-like but not depression-like behavior in mice as consequences of CSF modeling, were evaluated with Morris water maze (MWM), Novel object recognition (NOR), Open field test (OFT) and Forced swimming test (FST). In comparison with other sleep manipulations, this protocol minimizes the handling labors and maximizes the modeling efficiency. It produces stable phenotypes in young wild-type mice and can be potentially generated for a variety of research purposes.

Presented here is a protocol for chronic sleep fragmentation (CSF) model achieved by an electrically controlled orbital rotor, which could induce confirmed cognitive deficit and anxiety-like behavior in young wild-type mice. This model can be applied to explore the pathogenesis of chronic sleep disturbance and related disorders.

Um modelo de fragmentação crônica do sono usando rotor orbital vibrante para induzir déficit cognitivo e comportamento semelhante à ansiedade em camundongos jovens do tipo selvagem

Sleep disturbance is generally common in populations as a chronic disease or a complained event. Chronic sleep disturbance is proposed to be closely ...