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

Cette recherche a été financée par le programme de recherche intra-muros de la NIMH (ZIA MH00889). RMS a été également soutenue par une bourse de recherche postdoctorale FRAXA.

NOTE : Toutes les procédures ont été approuvées par le Comité de l’emploi et de National Institute of Mental Health Animal Care et effectués selon le National Institutes of Health Guidelines sur le soin et l’utilisation des animaux.
1. installation de privation de sommeil
<ol><li>Pinceaux doux permet de pousser doucement les souris pendant que vous dormez.<ol>
<li>Assurez-vous que le pinceau peut s’adapter à travers les barreaux de la mangeoire cage de souris pour s’assurer q...

<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>

Nous décrivons ici une méthode pour la privation de sommeil, douce manipulation, qui peut être utilisée dans les deux petits rongeurs et chez les adultes. Remise douce oblige les chercheurs à observer des souris pendant la durée d’une période de privation de sommeil et prod doucement les animaux lorsqu’ils sont inactifs ou contractions pour empêcher le sommeil. Études de privation de sommeil préalable ont vérifié au moyen d’enregistrements simultanés électroencéphalogramme (EEG) que manipulation douc...

Sommeil joue un rôle essentiel dans la formation de la plasticité et la synapse neuronale et élimination pendant le développement de cerveau1,2. Plus précisément, sommeil paradoxal (REM) est essentiel pour la formation des circuits synaptiques stables en renforçant les deux synapses spécifiques et élagage synaptique2. Privation de sommeil pharmacologique tôt dans la vie conduit à de nombreux déficits physiologiques et comportementaux3,4. En plus de sommeil pharmacologique privation, autres formes de privation de sommeil comme une agitation constante, forcé de locomotion, ou présentation d’un stimulus aversif ont été associés à la dépression-comme des symptômes à l’âge adulte, différent neurones activation des modes et des changements dans la continuité de temps et de sommeil sommeil pendant une période de récupération après privation5,6,7. Lorsque le sommeil est chroniquement limité chez les souris pendant plus de cinq semaines au cours du développement, des déficits comportementaux ont été observés après un mois de sommeil récupération8. Pris ensemble, ces études suggèrent que la perturbation du sommeil chez le rongeur néonatal peut affecter les habitudes de sommeil plus tard, le fonctionnement du cerveau et le comportement. Soulignant l’importance du sommeil pour le développement normal du cerveau chez l’être humain, de nombreux patients souffrant de troubles du développement neurologique, y compris les troubles du spectre autistique, complexe de sclérose tubéreuse de Bourneville, Syndrome de le X Fragile et d’Attention-Déficit/hyperactivité Trouble, des anomalies de rapport dans les habitudes de sommeil en plus une variété de déficits comportementaux1.
Vu le nombre de désordres neurodéveloppementaux qui sont associés à des habitudes de sommeil anormal, il est important de comprendre comment le manque de sommeil affecte le fonctionnement du cerveau et comportement. Cependant, malgré les données qui confirment l’importance du sommeil au cours du développement, la plupart des méthodologies qui restreignent ou privent les animaux d’activité de sommeil sur les adultes. Ces techniques incluent une variété de tests qui requièrent contraint l’activité locomotrice (p. ex., tapis de course constante), constante perturbation (p. ex., rotation bar balayeuse ou présentation d’objet roman continue) ou déranger l’animal au début de la Le sommeil paradoxal (p. ex., la plate-forme au-dessus de l’eau)9,10. Bien que ces méthodes aient démontré efficacement les problèmes liés au manque de sommeil chez l’adulte, aucune de ces méthodes sont appropriées pour les petits rongeurs à cause de leur mobilité réduite, voiture de sommeil haute et les effets néfastes du stress tôt dans la vie. En raison de l’importance du sommeil au cours du développement, il est essentiel de comprendre comment restreint de sommeil dans le comportement futur de chiots nouveau-nés affecte à l’âge adulte.
Une technique pour limiter le sommeil qui peut être utilisé chez les petits rongeurs est manipulation douce. Manipulation douce implique l’enquêteur interagissant directement avec les rongeurs que tout comportement au moment du sommeil est observée. Enquêteurs peuvent utiliser leurs mains, un pinceau ou la présentation d’objets nouveaux perturber physiquement couchés sur des rongeurs nouveau-nés8,10,11,12,13,14 . Sommeil peut être déterminé sur le plan comportemental en manque d’activité motrice, secousses myocloniques et fermeture des yeux (chez les petits rongeurs plus âgés). Validation de l’EEG confirme que perturber le sommeil avec manipulation douce basée sur ces comportements réduit le temps de sommeil total de 91 % en P12 rats14. Autres techniques qui ont été utilisés pour priver les jeunes souris incluent électrocution et la présentation de stimuli désagréables, mais ces techniques ne peuvent être utilisés pour la restriction chronique de sommeil et sont plus stressantes que manipulation douce7, 15. restriction chronique de sommeil peut également être exécutée via un protocole de secousse qui peut être utilisé avec ou sans implantation d’électrode EEG et de logiciels connexes, mais ce protocole n’empêche pas tous dormir et est donc moins contrôlée par rapport à manutention douce4,16. La capacité de restreindre ou de priver les souris de sommeil pendant plusieurs jours d’affilée sans équipement spécifique, logiciels ou l’implantation de l’électrode est un avantage de la technique de manipulation douce. Restriction de sommeil chronique via manipulation douce effectivement limite le sommeil chez les rongeurs nouveau-nés et produit une gamme de changements de comportement après le sommeil privation période8,11,12, 17.
Nous décrivons ici les résultats dans laquelle privation de sommeil chronique pendant trois heures par jour du jour après la naissance 5 (P5) au P42 altèrent de sociabilité de souris après une période de récupération de 30 jours dans lequel sommeil n’est pas perturbé. Ces résultats mettent en évidence les effets à long terme de restriction chronique de sommeil au cours du développement sur le comportement futur.

<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

Le sommeil est essentiel pour assurer le bon développement et plasticité neurale. En outre, les habitudes de sommeil anormal sont caractéristique de nombreux désordres neurodéveloppementaux. Restriction de sommeil chronique Comment étudier au cours du développement peut affecter le comportement adulte peut ajouter à notre compréhension de l’apparition de symptômes comportementaux des troubles du développement neurologique. Bien qu’il existe beaucoup de méthodes qui peut être utilisé pour restreindre le sommeil chez les rongeurs, y compris la locomotion forcée, perturbation constante, présentation d’un stimulus aversif ou choc électrique, beaucoup de ces méthodes sont très stressants et ne peuvent être utilisés chez les souris néonatales. Nous décrivons ici la manipulation douce, une technique de privation de sommeil qui peut être utilisée chroniquement tout au long de l’élaboration et à l’âge adulte pour réaliser la restriction de sommeil. Manipulation douce implique une surveillance étroite des souris tout au long de la période de privation de sommeil et oblige le chercheur à prod doucement les animaux lorsqu’ils sont inactifs ou affichent des comportements associés à dormir. Couplé avec les enregistrements EEG, manipulation douce pourrait servir à interrompre sélectivement une phase spécifique de sommeil comme le sommeil paradoxal (REM). La technique de manipulation douce est un outil puissant pour l’étude des effets de la restriction de sommeil chronique même chez les souris néonatales qui contourne les nombreuses procédures plus stressantes utilisées pour la privation de sommeil.

Afin d’étudier les effets de la restriction chronique de sommeil sur le comportement, nous dormons souris restreintes pendant trois heures tous les jours entre 11:00 et 14:00 de P5 à P428. Suite à la restriction de sommeil, les souris étaient laissés intacts pendant quatre semaines et a permis de récupérer de la restriction de sommeil. Souris ont été testées pour le comportement suivant la période de quatre semaines de récupération. Le test de compor...

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

Nous décrivons une technique de privation de sommeil dite douce manipulation où les enquêteurs prod doucement souris tout moment sommeil comportement est observé. Cette méthode est un outil puissant qui permet aux chercheurs d’étudier les effets que la restriction chronique de sommeil au cours du développement peut avoir sur le comportement et la physiologie du cerveau futures.

Privation de sommeil chronique chez la souris chiots au moyen de doux manutention

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

Cette méthode peut aider à répondre à des questions clés dans le domaine du sommeil telles que les effets de la restriction chronique du sommeil pendant le développement postnatal. Le principal avantage de cette technique est qu’elle est moins stressante que les autres techniques de privation de sommeil et peut facilement être utilisée tout au long du développement et se poursuivre jusqu’à l’âge adulte. Commencez par attribuer chaque cage soit au groupe de privation de sommeil, soit au groupe témoin.Assurez-vous que les souris ont accès à la nourriture et à l’eau ad libitum, et retirez tout aliment, literie ou objets d’enrichissement qui pourraient restreindre la vue des souris. Pour le groupe témoin, retirez le père de la cage d’éleveur le cinquième jour postnatal et laissez le barrage avec les chiots jusqu’au sevrage. Ensuite, déplacez les souris assignées au groupe témoin dans la salle de privation de sommeil et poussez doucement les souris avec un pinceau en continu pendant 10 minutes par jour pendant toute la durée de l’expérience.Après 10 minutes, retourner les souris dans la salle de détention de l’animal et ne pas les déranger. Commencez par retirer le père de la cage d’éleveur le cinquième jour postnatal et laissez le barrage avec les chiots jusqu’au sevrage. Déplacez quotidiennement les souris affectées au groupe de privation de sommeil dans la salle de privation de sommeil pendant le cycle de lumière où elles seront surveillées.Pendant ce temps, poussez doucement la souris avec un pinceau si on le soupçonne d’être endormi jusqu’à ce qu’une réponse soit observée. Alternativement, inverser les souris et pousser sur leur dos pour perturber le sommeil. Considérez qu’une souris dort si l’un des éléments suivants se produit : contractions, inactivité ou si les yeux sont fermés.Considérez les souris comme étant éveillées si elles se déplacent, essayant de se retourner après avoir été sur leur dos, ou se toilettant. Si le barrage couvre les chiots pendant la privation de sommeil, poussez-la doucement loin des chiots de sorte que la vue des chiots est sans restriction. Continuer avec le protocole de privation de sommeil jusqu’au point de terminaison de l’expérience.Lorsque la période de privation de sommeil de trois heures est terminée, remettre la literie et toute nourriture qui aurait pu être retirée et remettre le couvercle sur la cage. Enfin, retournez les cages dans la salle de détention des animaux jusqu’au lendemain de privation de sommeil. Après une période de rétablissement d’un mois, les résultats ont indiqué que dans le test de sociabilité, les femelles ayant restreint le sommeil passaient plus de temps à renifler la souris étrangère que les femelles témoins et les mâles restreints au sommeil avaient tendance à passer plus de temps à renifler l’objet que les mâles témoins.En outre, dans la phase de nouveauté sociale de l’essai, les souris femelles restreintes de sommeil ont passé une quantité accrue de temps reniflant la souris nouvelle que les contrôles femelles. Tout en essayant cette procédure, il est important de se rappeler de prêter attention et de répondre rapidement aux comportements pré-sommeil, en particulier chez les souris néonatales où le besoin de sommeil est élevé. Suivant cette procédure, d’autres méthodes comme le champ ouvert, rotarod, labyrinthe élevé plus, et les tests de comportement social peuvent être effectués afin de répondre à des questions supplémentaires comme les effets comportementaux à long terme de la restriction chronique du sommeil pendant le développement.

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.

Le groupe de congruence intermodale comme un moyen d&#39;obtenir une mesure comportementale objectif dans le caoutchouc main 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 ...

Recherche

Sciences du comportement

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.

L&#39;utilisation de la boîte de Puzzle comme un moyen d&#39;évaluer l&#39;efficacité d&#39;enrichissement du milieu

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.

Erreurs comme un moyen de réduire Impulsive choix d&#39;aliments

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.

Manipulation des épileptiforme Electrocorticograms (ECoGs) et du sommeil chez les rats et les souris par Acupuncture

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.

Neurodéveloppemental Test Reflex rats nouveau-nés Pups

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.

Non invasive, High-throughput détermination de la durée du sommeil chez les rongeurs

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.

Mesure Active et Passive docilité séparément chez les souris

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.

Reconnaissance d’objets nouveaux et emplacement de l’objet comportementale essais chez la souris sur un Budget

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.

Mesures automatisées de sommeil et de l’activité locomotrice dans Agassizii mexicain

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 modèle chronique de fragmentation du sommeil utilisant rotor orbital vibrant pour induire le déficit cognitif et le comportement anxieux chez les jeunes souris de type sauvage

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