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 investigación fue financiada por el programa de investigación intramuros del NIMH (ZIA MH00889). RMS también fue apoyado por una Beca Postdoctoral de FRAXA.

Nota: Todos los procedimientos fueron aprobados por el nacional Instituto of Mental Salud Animal Care y uso y conformidad con directrices para los institutos nacionales de salud en el cuidado y uso de animales.
1. configuración de la privación de sueño
<ol><li>Usar pinceles blandos para suavemente empujar ratones durante el sueño.<ol>
<li>Asegúrese de que el pincel puede caber a través de las barras del comedero de la jaula del ratón para asegurarse de que los ratones tienen acceso a ...

<ol>
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A. <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+in+rats:+Effects+on+EEG+power+spectra,+vigilance+states,+and+cortical+temperature.">Sleep deprivation in rats: Effects on EEG power spectra, vigilance states, and cortical temperature.</a> Am J Physiol. 261 (1 Pt 2), R198-R208 (1991).</li><li>Araujo, P., Coelho, C. A., Oliveira, M. G., Tufik, S., Andersen, M. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neonatal+sleep+restriction+increases+nociceptive+sensitivity+in+adolescent+mice.">Neonatal sleep restriction increases nociceptive sensitivity in adolescent mice.</a> Pain Physician. 21 (2), E137-E148 (2018).</li><li>Araujo, P., Tufik, S., Anderson, M. 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. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=An+instrumental+method+for+long-term+continuous+REM+sleep+deprivation+of+neonatal+rats.">An instrumental method for long-term continuous REM sleep deprivation of neonatal rats.</a> Sleep. 23 (2), 175-183 (2000).</li><li>Xu, Z. Q., Gao, C. Y., Fang, C. Q., Zhou, H. D., Jiang, X. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+mechanism+and+characterization+of+learning+and+memory+impairment+in+sleep-deprived+mice.">The mechanism and characterization of learning and memory impairment in sleep-deprived mice.</a> Cell Biochem Biophys. 58 (3), 137-140 (2010).</li><li>Barriga, C., Martin, M. I., Tabla, R., Ortega, E., Rodriguez, A. 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. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+union+of+the+state:+Myoclonic+twitching+is+coupled+with+nuchal+muscle+atonia+in+infant+rats.">The union of the state: Myoclonic twitching is coupled with nuchal muscle atonia in infant rats.</a> Behav Neurosci. 116 (5), 912-917 (2002).</li></ol>

Aquí se describe un método para la privación del sueño, suave manejo, que puede utilizarse en ambos roedores cachorros y adultos. Entrega suave requiere que los investigadores observar los ratones durante un período de privación de sueño y suavemente prod los animales cuando están inactivos o temblor para evitar el sueño. Estudios de privación de sueño previa han comprobado por medio de grabaciones simultáneo del electroencefalograma (EEG) que evita la manipulación suave tanto REM y no - REM NREM sueño<sup ...

Sueño juega un papel fundamental en la formación de sinapsis y la plasticidad neuronal y eliminación durante el cerebro desarrollo1,2. En concreto, movimientos oculares rápidos (REM) es esencial para la formación de circuitos sinápticos estables por tanto fortalecimiento de la sinapsis específicas y poda sináptica2. La privación del sueño farmacológico temprano en la vida lleva a muchos déficits fisiológicos y de comportamiento3,4. Además de dormir farmacológica privación, otras formas de privación de sueño como agitación constante, forzado locomoción, o presentación de un estímulo aversivo se han asociado con síntomas como la depresión en la edad adulta, diferentes nervios patrones de activación y cambios en la continuidad de tiempo y sueño de sueño durante un período de recuperación después de privación5,6,7. Cuando sueño es crónica en ratones durante más de cinco semanas durante el desarrollo, déficits conductuales fueron observados después de un mes de sueño de recuperación8. Tomados juntos, estos estudios sugieren que la interrupción del sueño en el roedor neonatal puede afectar más adelante patrones del sueño, función cerebral y comportamiento. La importancia del sueño para el desarrollo normal del cerebro en los seres humanos, muchos pacientes con trastornos del desarrollo neurológico, incluyendo trastornos del espectro autista, complejo de esclerosis tuberosa, síndrome de X frágil y déficit de atención con hiperactividad Desorden, las anormalidades en patrones de sueño, además de una gran variedad de déficits conductuales1informe.
Dado el número de trastornos del neurodesarrollo que se asocian a patrones de sueño anormales, es importante entender cómo la falta de sueño afecta el comportamiento y la función cerebral. Sin embargo, a pesar de los datos que apoyan la importancia de dormir durante el desarrollo, más metodologías que restringen o privar a los animales del foco del sueño en adultos. Estas técnicas incluyen una variedad de pruebas que requieren obligados actividad locomotora (por ejemplo, constante caminadora), interrupción constante (por ejemplo, girando la barra de barrendero o presentación objeto novela continua) o molestar a los animales en el inicio de El sueño REM (por ejemplo, la plataforma sobre el agua)9,10. Aunque estos métodos han demostrado eficacia los problemas relacionados con la privación del sueño en adultos, ninguno de estos métodos son apropiado para crías de roedores debido a su limitada movilidad, sueño alto en coche y los efectos adversos del estrés temprano en la vida. Debido a la importancia del sueño durante el desarrollo, es fundamental para entender el sueño como restringido en el comportamiento futuro de los cachorros neonatales afecta durante la edad adulta.
Una técnica para restringir el sueño que puede ser utilizado en crías de roedores es un manejo suave. Manipulación suave implica el investigador interactuar directamente con los roedores que se observa cualquier comportamiento en tiempo de sueño. Los investigadores pueden utilizar sus manos, un pincel o la presentación de objetos novedosos para alterar físicamente el sueño en roedores neonatales8,10,11,12,13,14 . Sueño puede determinarse su comportamiento por la falta de actividad motora, espasmos mioclónicos y encierro del ojo (en crías de roedores mayores). Validación de EEG confirma que perturban el sueño con la dirección apacible basado en estos comportamientos reduce el tiempo de sueño total en un 91% en P12 ratas14. Otras técnicas que se han utilizado para privar a ratones jóvenes incluyen descargas eléctricas y presentación de los estímulos desagradables, pero estas técnicas no pueden utilizarse para restricción de sueño crónica y son más estresantes que una manipulación suave7, 15. restricción de sueño crónica también puede lograrse a través de un protocolo de agitación que puede utilizarse con o sin implantación de electrodos de EEG y los programas informáticos asociados, pero este Protocolo no impide dormir todos y por lo tanto es menos controlada en comparación con un manejo suave4,16. La capacidad de restringir o privar a los ratones de sueño durante varios días en una fila sin equipos específicos, programas informáticos o implantación de electrodo es un beneficio de la técnica de manipulación suave. Restricción del sueño crónica mediante manipulación suave efectivamente limita el sueño en roedores neonatales y produce una variedad de cambios en el comportamiento tras el sueño privación período8,11,12, 17.
Aquí, Describimos resultados que privación de sueño crónica durante tres horas por día del día postnatal 5 (P5) a P42 afecta significativamente la sociabilidad de los ratones después de un período de recuperación de 30 días en el que sueño no es perturbado. Estos resultados destacan los efectos a largo plazo de la restricción del sueño crónica durante el desarrollo de comportamiento 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

El sueño es fundamental para el correcto desarrollo y plasticidad neuronal. Por otra parte, los patrones de sueño anormales son características de muchos trastornos del neurodesarrollo. Estudiar cómo crónica restricción del sueño durante el desarrollo puede afectar comportamiento adulto puede aumentar nuestra comprensión de la aparición de síntomas conductuales de trastornos del neurodesarrollo. Si bien hay muchos métodos que pueden utilizarse para restringir el sueño en roedores incluyendo locomoción forzada, interrupción constante, presentación de un estímulo aversivo o descarga eléctrica, muchos de estos métodos son muy estresantes y no pueden usarse en ratones neonatales. Aquí, describimos la manipulación suave, una técnica de privación de sueño que puede utilizarse crónicamente a lo largo de desarrollo y en la edad adulta para lograr la restricción del sueño. Manipulación suave consiste en una observación más cercana de los ratones durante todo el período de privación de sueño y requiere al investigador suavemente prod los animales cuando están inactivos o mostrar comportamientos asociados con el sueño. Junto con las grabaciones de EEG, manejo suave podría utilizarse para interrumpir selectivamente una fase específica del sueño como el sueño de movimientos oculares rápidos (MOR). La técnica de manejo suave es una poderosa herramienta para el estudio de los efectos de restricción del sueño crónica incluso en ratones neonatales que evita muchos de los procedimientos más estresantes para la privación del sueño.

Para investigar los efectos de la restricción del sueño crónica sobre el comportamiento, dormimos ratones restringidas durante tres horas todos los días entre las 11:00 y 14:00 de P5 a P428. A raíz de la restricción del sueño, ratones quedaban imperturbados durante cuatro semanas y permite recuperarse de restricción del sueño. Ratones fueron probados por su comportamiento tras el período de recuperación de cuatro semanas. La prueba de comportamiento soci...

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

Describimos una técnica de privación de sueño conocida como donde investigadores suavemente producciën ratones cualquier momento comportamiento de sueño se observa un manejo suave. Este método es una poderosa herramienta que permite a los investigadores estudiar los efectos que puede tener restricción del sueño crónica en todo el desarrollo sobre el comportamiento y la fisiología cerebral futuro.

Privación de sueño crónica en ratón cachorros por medio de suave manejo

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

Este método puede ayudar a responder preguntas clave en el campo del sueño, como los efectos de la restricción crónica del sueño durante el desarrollo posnatal. La principal ventaja de esta técnica es que es menos estresante que otras técnicas de privación del sueño y se puede utilizar fácilmente durante todo el desarrollo y continuar en la edad adulta. Comience asignando cada jaula a la privación del sueño o al grupo de control.Asegúrese de que los ratones tengan acceso a alimentos y agua ad libitum, y retire cualquier alimento, ropa de cama o objetos de enriquecimiento que puedan restringir la vista de los ratones. Para el grupo de control, retire al señor de la jaula del criador el día cinco postnatal y salga de la presa con los cachorros hasta el destete. A continuación, mueva los ratones asignados al grupo de control a la sala de privación del sueño y prodly los ratones con un pincel continuamente durante 10 minutos al día durante la duración del experimento.Después de 10 minutos, devolver los ratones a la sala de retención del animal y no molestarlos. Comience retirando el señor de la jaula del criador el día cinco postnatal y deje la presa con los cachorros hasta el destete. Mueva los ratones asignados al grupo de privación del sueño a la sala de privación del sueño diariamente durante el ciclo de luz donde serán monitoreados.Durante este tiempo, coloque suavemente el ratón con un pincel si se sospecha que está dormido hasta que se observe una respuesta. Alternativamente, invierta ratones y empuje sobre sus espaldas para interrumpir el sueño. Considere que un ratón está dormido si ocurre alguna de las siguientes situaciones: espasmos, inactividad o si los ojos están cerrados.Considere que los ratones están despiertos si se están moviendo, tratando de voltear después de estar de espaldas o acicalándose. Si la presa está cubriendo a los cachorros durante la privación del sueño, aleje suavemente de los cachorros para que la vista de los cachorros no esté restringida. Continúe con el protocolo de privación de sueño hasta el punto final del experimento.Cuando se complete el período de tres horas de privación del sueño, coloque la ropa de cama y cualquier alimento que pueda haber sido retirado de nuevo y vuelva a colocar la tapa en la jaula. Finalmente, devuelva las jaulas a la sala de retención de animales hasta el día siguiente de la privación del sueño. Después de un período de recuperación de un mes, los resultados indicaron que en la prueba de sociabilidad, las hembras restringidas de sueño pasaban más tiempo olfateando al ratón extraño que las hembras de control y los machos con restricciones de sueño tenían una tendencia a pasar más tiempo olfateando el objeto en comparación con los machos de control.Además, en la fase de novedad social de las pruebas, los ratones hembra restringidos por el sueño pasaban una mayor cantidad de tiempo olfateando el ratón novedoso que los controles femeninos. Al intentar este procedimiento, es importante recordar prestar atención y responder rápidamente a los comportamientos previos al sueño, especialmente en ratones neonatales donde la necesidad de sueño es alta. Después de este procedimiento, se pueden realizar otros métodos como campo abierto, rotarod, laberinto elevado más, y pruebas de comportamiento social con el fin de responder a preguntas adicionales como los efectos conductuales a largo plazo de la restricción crónica del sueño durante el desarrollo.

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.

El Grupo de Congruencia crossmodal como medio para obtener una medida del comportamiento objetivo de la Guía Caucho mano 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 ...

Investigación

Comportamiento

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.

El uso de la caja de rompecabezas como un medio para evaluar la eficacia de Enriquecimiento 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.

Errores como un medio de reducir impulsivo Choice Food

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.

La manipulación de epileptiforme Electrocorticograms (ECoGs) y del sueño en ratas y ratones 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.

Neurodesarrollo pruebas de reflejos neonatales en crías de rata

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.

Determinación no invasiva, de alto rendimiento de la duración del sueño en 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.

Medición activa y pasiva mansedumbre por separado en ratones

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.

Reconocimiento de objetos nuevos y ubicación del objeto conductual en ratones en un presupuesto

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.

Mediciones automatizadas de sueño y actividad locomotriz en mexicano Cavefish

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.

Un modelo crónico de fragmentación del sueño usando rotor orbital vibratorio para inducir déficit cognitivo y comportamiento similar a la ansiedad en ratones jóvenes de tipo salvaje

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