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

Questa ricerca è stata finanziata dal programma di ricerca intramurale del NIMH (ZIA MH00889). RMS è stato supportato anche da una borsa di studio post-dottorato di FRAXA.

Nota: Tutte le procedure erano approvate dal comitato di uso e National Institute di mentale salute Animal Care ed eseguite secondo gli istituti nazionali di salute orientamenti sull'uso degli animali e manutenzione.
1. installazione di privazione di sonno
<ol><li>Usare i pennelli morbidi per colpire delicatamente topi durante il sonno.<ol>
<li>Assicurarsi che il pennello può adattarsi attraverso le sbarre dell'alimentatore del mouse gabbia per garantire che i topi hanno accesso al cibo e a...

<ol>
	<li>Picchioni, D., Reith, R. M., Nadel, J. L., Smith, C. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sleep,+plasticity+and+the+pathophysiology+of+neurodevelopmental+disorders:+the+potential+roles+of+protein+synthesis+and+other+cellular+processes.">Sleep, plasticity and the pathophysiology of neurodevelopmental disorders: the potential roles of protein synthesis and other cellular processes.</a> Brain Sci. 4 (1), 150-201 (2014).</li><li>Li, W., Ma, L., Yang, G., Gan, W. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=REM+sleep+selectively+prunes+and+maintains+new+synapses+in+development+and+learning.">REM sleep selectively prunes and maintains new synapses in development and learning.</a> Nat Neurosci. 20 (3), 427-437 (2017).</li><li>Peirano, P. D., Algarin, C. 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G., Morrissette, R., Heller, H. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effects+of+sleep+deprivation+in+neonatal+rats.">Effects of sleep deprivation in neonatal rats.</a> Am J Physiol. 275 (1 Pt 2), R148-R157 (1998).</li><li>Todd, W. D., Gibson, J. L., Shaw, C. S., 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=Brainstem+and+hypothalamic+regulation+of+sleep+pressure+and+rebound+in+newborn+rats.">Brainstem and hypothalamic regulation of sleep pressure and rebound in newborn rats.</a> Behav Neurosci. 124 (1), 69-78 (2010).</li><li>Sare, R. M., Levine, M., Hildreth, C., Picchioni, D., Smith, C. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Chronic+sleep+restriction+during+development+can+lead+to+long-lasting+behavioral+effects.">Chronic sleep restriction during development can lead to long-lasting behavioral effects.</a> Physiol Behav. 155, 208-217 (2016).</li><li>Colavito, V., 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=Experimental+sleep+deprivation+as+a+tool+to+test+memory+deficits+in+rodents.">Experimental sleep deprivation as a tool to test memory deficits in rodents.</a> Front Syst Neurosci. 7, 106 (2013).</li><li>Franken, P., Dijk, D. J., Tobler, I., Borbely, A. 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>

Qui descriviamo un metodo per la privazione di sonno, movimentazione, che può essere utilizzato in entrambi cuccioli roditore e negli adulti di gentile. Consegna dolci richiede ai ricercatori di osservare i topi per la durata di un periodo di privazione di sonno e prod delicatamente gli animali quando sono inattivi o contrazioni per impedire il sonno. Gli studi di privazione di sonno precedente hanno verificato per mezzo di registrazioni simultanee elettroencefalogramma (EEG) che impedisce la manipolazione delicata sia ...

Il sonno gioca un ruolo critico nella formazione di plasticità e sinapsi neuronale e l'eliminazione durante lo sviluppo del cervello1,2. In particolare, sonno veloce del movimento oculare (REM) è essenziale per la formazione di circuiti sinaptici stabile attraverso entrambi rafforzamento di specifiche sinapsi e sinaptica potatura2. Privazione del sonno farmacologico presto nella vita conduce a molti deficit fisiologici e comportamentali3,4. Oltre al sonno farmacologico privazione, altre forme di privazione del sonno come agitazione costante, costretto locomozione o presentazione di uno stimolo avversivo sono stati associati con la depressione-come i sintomi durante l'età adulta, diverso neurale pattern di attivazione e cambiamenti nella continuità di tempo e il sonno sonno durante un periodo di recupero dopo privazione5,6,7. Quando il sonno è cronicamente limitato nei topi per cinque settimane durante lo sviluppo, deficit comportamentistici sono stati osservati dopo un mese di sonno recupero8. Presi insieme, questi studi suggeriscono che la rottura di sonno nel roditore neonatale possa influenzare successivi modelli di sonno, la funzione del cervello e comportamento. Evidenziando l'importanza del sonno per lo sviluppo normale del cervello in esseri umani, molti pazienti con disturbi dello sviluppo neurologico, tra cui disturbi dello spettro autistico, sclerosi tuberosa complessa, sindrome da X Fragile e Attention-Deficit/Hyperactivity Disordine, le anomalie di relazione nel ritmo del sonno oltre ad una varietà di deficit comportamentistici1.
Dato il numero di disturbi dello sviluppo neurologico che sono associati con modelli di sonno anormale, è importante capire come la mancanza di sonno influisce comportamento e funzione del cervello. Tuttavia, nonostante i dati che supportano l'importanza del sonno durante lo sviluppo, la maggior parte delle metodologie che limitano o privano gli animali di messa a fuoco di sonno sugli adulti. Queste tecniche comprendono una varietà di prove che richiedono forzati attività locomotrice (ad es., costante tapis roulant), costante interruzione (ad es., rotante barra spazzatrice o presentazione oggetto continuo di romanzo) o disturbare l'animale al momento della comparsa di Sonno di rem (ad es., piattaforma sull'acqua)9,10. Sebbene questi metodi hanno dimostrato in modo efficace i problemi legati alla privazione di sonno in adulti, nessuno di questi metodi sono appropriati per roditori cuccioli a causa della loro mobilità limitata, unità alto sonno e gli effetti negativi dello stress presto nella vita. Data l'importanza del sonno durante lo sviluppo, è fondamentale per capire come limitato sonno nel comportamento futuro di cuccioli neonatale colpisce durante l'età adulta.
Una tecnica per limitare il sonno che può essere utilizzato nei cuccioli roditore è una manipolazione delicata. Manipolazione delicata coinvolge l'investigatore che interagiscono direttamente con i roditori che qualsiasi comportamento di sonno ora è osservato. Gli investigatori possono utilizzare le loro mani, un pennello o presentazione di nuovi oggetti per fisicamente interrompe il sonno in roditori neonatale8,10,11,12,13,14 . Sonno può essere relativamente al comportamento determinato dalla mancanza di attività motoria, spasmi mioclonici e chiusura degli occhi (in più vecchi cuccioli roditori). Convalida di EEG conferma che interrompere il sonno con delicatezza basato su questi comportamenti riduce il tempo totale di sonno di 91% nel P12 ratti14. Altre tecniche che sono state utilizzate per privare i giovani topi includono scosse elettriche e presentazione di stimoli sgradevoli, ma queste tecniche non possono essere utilizzate per la restrizione di sonno cronica e sono più stressante di manipolazione delicata7, 15. la restrizione di sonno cronica può essere effettuata anche tramite un protocollo di agitazione che può essere utilizzato con o senza l'impianto di elettrodi EEG ed il software associato, ma questo protocollo non impediscono il sonno tutti e quindi è meno controllato rispetto a manipolazione delicata4,16. La capacità di limitare o privare topi di sonno per molti giorni di fila senza attrezzature specifiche, software per computer o l'impianto dell'elettrodo è un vantaggio della tecnica di manipolazione delicata. Restrizione di sonno cronica tramite manipolazione delicata efficacemente limita il sonno nei roditori neonatale e produce una varietà di cambiamenti comportamentali seguendo il sonno privazione periodo8,11,12, 17.
Qui, descriviamo i risultati in cui privazione di sonno cronica per tre ore al giorno dal giorno postnatale 5 (P5) al P42 influisce in modo significativo la socievolezza dei topi dopo un periodo di recupero di 30 giorni in cui il sonno non è disturbato. Questi risultati evidenziano gli effetti a lungo termine della restrizione di sonno cronica durante lo sviluppo il 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

Sonno è fondamentale per il corretto sviluppo e plasticità neurale. Inoltre, modelli di sonno anormale sono caratteristici di molti disturbi dello sviluppo neurologico. Restrizione di sonno cronica come studiare durante lo sviluppo possa influire sul comportamento adulto può aggiungere alla nostra comprensione della comparsa di sintomi comportamentali di disturbi dello sviluppo neurologico. Mentre ci sono molti metodi che possono essere utilizzati per limitare il sonno nei roditori tra cui locomozione forzata, costante interruzione, presentazione di uno stimolo avversivo, o scosse elettriche, molti di questi metodi sono molto stressanti e non può essere utilizzati in topi neonatali. Qui, descriviamo manipolazione delicata, una tecnica di privazione di sonno che può essere utilizzata cronicamente durante lo sviluppo e nell'età adulta per raggiungere la restrizione di sonno. Manipolazione delicata comporta la stretta osservazione dei topi per tutto il periodo di privazione di sonno e richiede al ricercatore di prod delicatamente gli animali quando sono inattivi o visualizzare i comportamenti associati al sonno. Accoppiato con le registrazioni di EEG, manipolazione delicata potrebbe essere utilizzata per distruggere selettivamente una specifica fase del sonno come sonno veloce del movimento oculare (REM). La tecnica di manipolazione delicata è un potente strumento per lo studio degli effetti della restrizione di sonno cronica anche in topi neonatali che aggira molte delle procedure più stressante utilizzate per la privazione di sonno.

Per studiare gli effetti della restrizione di sonno cronica sul comportamento, dormiamo limitati topi per tre ore ogni giorno tra le 11:00 e 14:00 da P5 a P428. Dopo la restrizione di sonno, topi sono stati lasciati indisturbati per quattro settimane e ha permesso di recuperare dalla restrizione di sonno. Topi sono stati testati per un comportamento dopo il periodo di recupero di quattro settimane. La prova di comportamento sociale tre-alloggiamento è stata usata ...

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

Descriviamo una tecnica di privazione di sonno conosciuta come manipolazione delicata dove gli investigatori delicatamente prod topi qualsiasi momento viene osservato il comportamento di sonno. Questo metodo è un potente strumento che permette ai ricercatori di studiare gli effetti che la restrizione di sonno cronica nel corso dello sviluppo può avere sul comportamento e sulla fisiologia futuro cervello.

Deprivazione di sonno cronica nel topo cuccioli per mezzo di Gentle Handling

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

Questo metodo può aiutare a rispondere a domande chiave nel campo del sonno come gli effetti della restrizione cronica del sonno durante lo sviluppo postnatale. Il vantaggio principale di questa tecnica è che è meno stressante di altre tecniche di privazione del sonno e può essere facilmente utilizzato durante lo sviluppo e continuare fino all'età adulta. Inizia assegnando ogni gabbia alla privazione del sonno o al gruppo di controllo.Assicurarsi che i topi abbiano accesso a cibo e acqua ad libitum e rimuovere qualsiasi cibo, biancheria da letto o oggetti di arricchimento che potrebbero limitare la vista dei topi. Per il gruppo di controllo, rimuovere il sire dalla gabbia dell'allevatore il quinto giorno postnatale e lasciare la diga con i cuccioli fino allo svezzamento. Quindi, spostare i topi assegnati al gruppo di controllo nella stanza di privazione del sonno e pungere delicatamente i topi con un pennello continuamente per 10 minuti al giorno per tutta la durata dell'esperimento.Dopo 10 minuti, riportare i topi nella stanza di detenzione dell'animale e non disturbarli. Inizia rimuovendo il sire dalla gabbia dell'allevatore il quinto giorno postnatale e lascia la diga con i cuccioli fino allo svezzamento. Spostare quotidianamente i topi assegnati al gruppo di privazione del sonno nella stanza di privazione del sonno durante il ciclo di luce in cui verranno monitorati.Durante questo periodo, pungere delicatamente il mouse con un pennello se sospettato di dormire fino a quando non viene osservata una risposta. In alternativa, inverti i topi e spingi sulla schiena per interrompere il sonno. Considera un mouse addormentato se si verifica una delle seguenti contrazioni: contrazioni, inattività o se gli occhi sono chiusi.Considera i topi svegli se si muovono, cercano di capovolgersi dopo essere stati sulla schiena o pulirsi. Se la diga copre i cuccioli durante la privazione del sonno, allontanarla delicatamente dai cuccioli in modo che la vista dei cuccioli sia illimitata. Continuare con il protocollo di privazione del sonno fino all'endpoint dell'esperimento.Quando il periodo di tre ore di privazione del sonno è completo, metti la biancheria da letto e qualsiasi cibo che potrebbe essere stato rimosso e rimette il coperchio sulla gabbia. Infine, riportare le gabbie nella stanza di detenzione degli animali fino al giorno successivo di privazione del sonno. Dopo un periodo di recupero di un mese, i risultati hanno indicato che nel test di socialità, le femmine con restrizioni di sonno passavano più tempo a fiutare il topo straniero rispetto a controllare le femmine e i maschi con restrizioni di sonno avevano la tendenza a passare più tempo a annusare l'oggetto rispetto ai maschi di controllo.Inoltre, nella fase di novità sociale dei test, i topi femmine con restrizioni di sonno hanno trascorso una maggiore quantità di tempo a fiutare il nuovo topo rispetto ai controlli femminili. Durante il tentativo di questa procedura, è importante ricordare di prestare attenzione e rispondere prontamente ai comportamenti pre-sonno, specialmente nei topi neonatale in cui il bisogno di sonno è elevato. Seguendo questa procedura, altri metodi come campo aperto, rotarod, elevato più labirinto e test del comportamento sociale possono essere eseguiti al fine di rispondere a domande aggiuntive come gli effetti comportamentali a lungo termine della restrizione cronica del sonno durante lo sviluppo.

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.

La Task Congruenza crossmodali come un mezzo per ottenere una misura comportamentale Obiettivo della mano di gomma Illusion Paradigma

The rubber hand illusion (RHI) is a popular experimental paradigm. Participants view touch on an artificial rubber hand while the participants' own hidden ...

Ricerca

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.

L&#39;uso del Puzzle Box come un mezzo per valutare l&#39;efficacia di arricchimento ambientale

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.

Errori come un mezzo per ridurre impulsivo scelta alimentare

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.

Manipolazione di epilettiforme Electrocorticograms (ECoGs) e sonno in ratti e topi con l&#39;agopuntura

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.

Neurologico Reflex Test in neonatali Rat 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.

Determinazione non invasiva, High-throughput di durata di sonno in roditori

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.

Attiva e passiva docilità separatamente nei topi di misura

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.

Riconoscimento di oggetti nuovi e oggetto posizione comportamentali test in topi con un budget limitato

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.

Misure automatizzate di sonno e di attività locomotrice in caverna messicano

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 modello di frammentazione del sonno cronico che utilizza il rotore orbitale vibrante per indurre deficit cognitivo e comportamento simile all'ansia nei giovani topi di tipo selvaggio

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