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

Dit onderzoek werd gefinancierd door de intramurale Research Program van de NIMH (ZIA MH00889). RMS werd ook ondersteund door een FRAXA Postdoctoral Fellowship.

Opmerking: Alle procedures werden goedgekeurd door het nationale Instituut voor geestelijke gezondheid Animal Care en gebruik Comité en uitgevoerd overeenkomstig de nationale instituten van gezondheid richtsnoeren inzake de zorg en het gebruik van dieren.
1. slaap deprivatie Setup
<ol><li>Gebruik zachte schilderskwast zachtjes steken muizen als in slaap.<ol>
<li>Zorg ervoor dat het penseel kan passen door de tralies van de muis kooi feeder om ervoor te zorgen dat muizen toegang tot voedsel ...

<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. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sleep+in+brain+development.">Sleep in brain development.</a> Biol Res. 40 (4), 471-478 (2007).</li><li>Mirmiran, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effects+of+experimental+suppression+of+active+(REM)+sleep+during+early+development+upon+adult+brain+and+behavior+in+the+rat.">Effects of experimental suppression of active (REM) sleep during early development upon adult brain and behavior in the rat.</a> Brain Res. 283 (2-3), 277-286 (1983).</li><li>Feng, P., Ma, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Instrumental+REM+sleep+deprivation+in+neonates+leads+to+adult+depression-like+behaviors+in+rats.">Instrumental REM sleep deprivation in neonates leads to adult depression-like behaviors in rats.</a> Sleep. 26 (8), 990-996 (2003).</li><li>Frank, M. 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>

Hier beschrijven we een methode voor slaaptekort, zachte behandeling, die kan worden gebruikt in zowel knaagdier pups en bij volwassenen. Zachte inleveren vereist onderzoekers te observeren van muizen voor de duur van een periode van slaap deprivatie en zachtjes prod de dieren wanneer ze inactief of twitching ter voorkoming van slaap. Voorafgaande slaap deprivatie studies hebt gecontroleerd door middel van gelijktijdige electroencephalogram (EEG) opnamen dat zachte behandeling beide REM voorkomt en niet REM NREM - slaap<...

Slaap speelt een cruciale rol in de neuronale plasticiteit en synapse formatie en eliminatie tijdens1,2van de ontwikkeling van de hersenen. Rapid eye movement (REM) slaap is met name essentieel voor het vormen van stabiele synaptic circuits door beide versterking van specifieke synapsen en synaptische snoeien2. Farmacologische slaaptekort vroeg in het leven leidt tot veel fysiologische en gedragsmatige tekorten3,4. Naast farmacologische slaap deprivatie, andere vormen van slaaptekort zoals constante schudden, gedwongen motoriek of presentatie van een aversieve stimulus zijn geassocieerd met depressie-achtige symptomen tijdens volwassenheid, verschillende neurale activering patronen, en veranderingen in slaap tijd en slaap continuïteit tijdens een herstelperiode na ontbering5,6,7. Wanneer slaap het chronisch in muizen voor meer dan vijf weken tijdens de ontwikkeling beperkt is, de gedrags tekorten werden waargenomen na een maand van de slaap herstel8. Samen genomen, suggereren deze studies dat de verstoring van de slaap in de neonatale knaagdier later slaappatronen, hersenfunctie en -gedrag kan beïnvloeden. Wijzen op het belang van de slaap voor de ontwikkeling van de normale hersenen bij de mens, vele patiënten met neurologische aandoeningen, met inbegrip van Autisme Spectrum stoornissen, Tubereuze Sclerose Complex, fragiele X-syndroom en aandacht-Deficit/Hyperactivity Obsessieve compulsieve stoornis, verslag abnormaliteiten in slaappatronen naast allerlei gedrags tekorten1.
Gezien het aantal neurologische aandoeningen die gekoppeld aan abnormale slaappatronen zijn, is het belangrijk om te begrijpen hoe gebrek aan slaap beïnvloedt hersenfunctie en gedrag. Echter, ondanks de gegevens die het belang van slaap tijdens de ontwikkeling, ondersteunen de meeste methodologieën die te beperken of te ontnemen van slaap focus op volwassenen dieren. Deze technieken omvatten allerlei tests waarvoor gedwongen motorische activiteit (bijvoorbeeld, constante loopband), constante verstoring (bijvoorbeeld roteren Veger bar of voortdurende roman object presentatie), of het verstoren van het dier aan het begin van REM-slaap (bijvoorbeeld platform waterbeheersing)9,10. Hoewel deze methoden is de problemen in verband met slaaptekort bij volwassenen effectief gebleken, zijn geen van deze methoden geschikt voor knaagdieren pups vanwege hun beperkte mobiliteit, hoge slaap station en de schadelijke effecten van stress vroeg in het leven. Vanwege het belang van slaap tijdens de ontwikkeling is het essentieel om te begrijpen hoe beperkt slaap in neonatale pups beïnvloedt toekomstig gedrag tijdens volwassenheid.
Een techniek om te slapen, die kan worden gebruikt in knaagdier pups beperken is zachte behandeling. Zachte behandeling omvat de onderzoeker directe wisselwerking met de knaagdieren die ieder tijd slaap gedrag wordt waargenomen. Onderzoekers kunnen gebruiken hun handen, een penseel of presentatie van de nieuwe objecten fysiek verstoort slaap in neonatale knaagdieren8,10,11,12,13,14 . Slaap kan gedragsgestoorde worden bepaald door gebrek aan motorische activiteit, myoclonische spiertrekkingen en oog sluiting (in oudere knaagdier pups). EEG validatie bevestigt dat slaap verstoren met zachte behandeling op basis van deze gedragingen vermindert de totale slaaptijd met 91% in P12 ratten14. Andere technieken die zijn gebruikt om het ontnemen van de jonge muizen omvatten elektrische schok en presentatie van onaangename stimuli, maar deze technieken kunnen niet worden gebruikt voor chronische slaap beperking en zijn meer stress dan zachte behandeling7, 15. chronische slaap beperking kan ook worden bereikt door een schudden protocol dat kan worden gebruikt met of zonder EEG elektrode implantatie en bijbehorende computersoftware, maar dit protocol belet niet alle slaap en is daarom minder gecontroleerd ten opzichte van zachte behandeling4,16. De mogelijkheid om te beperken of te ontnemen van muizen van de slaap voor vele dagen in een rij zonder specifieke apparatuur, computersoftware of elektrode implantatie is een voordeel van de zachte behandeling techniek. Chronische slaap beperking via zachte behandeling effectief beperkt slaap in neonatale knaagdieren en produceert een scala aan gedragsveranderingen na de slaap deprivatie periode8,11,12, 17.
Hier beschrijven we resultaten waarin Chronisch slaaptekort gedurende drie uur per dag van postnatale dag 5 (P5) t P42 heeft grote invloed op gezelligheid van muizen na een herstelperiode van 30 dagen waarin slaap niet wordt verstoord. Deze resultaten wijzen op de lange termijn effecten van chronische slaap beperking tijdens ontwikkeling op toekomstig gedrag.

<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

Slaap is essentieel voor de goede ontwikkeling en Neurale plasticiteit. Bovendien, abnormale slaappatronen zijn kenmerkend voor veel neurologische aandoeningen. Bestuderen hoe chronische beperking van de slaap tijdens de ontwikkeling van invloed kan zijn op de volwassen gedrag kan toevoegen aan ons begrip van de opkomst van gedrags symptomen van neurologische aandoeningen. Hoewel er vele methoden die kunnen worden gebruikt om te slapen in knaagdieren, met inbegrip van gedwongen motoriek, verstoring van de constante, presentatie van een aversieve stimulus, of een elektrische schok te beperken, veel van deze methoden zijn zeer belastend en kunnen niet worden gebruikt bij neonatale muizen. We beschrijven hier, zachte behandeling, een slaap deprivatie techniek die kan gebruikt worden chronisch in de gehele ontwikkeling en in volwassenheid tot beperking van de slaap. Zachte behandeling omvat nauwkeurige observatie van de muizen gedurende de ontbering slaap en vereist de onderzoeker te zachtjes prod de dieren wanneer ze inactief worden of weer gedrag geassocieerd met slapen. In combinatie met EEG-opnamen, kan zachte behandeling worden gebruikt om een specifieke fase van de slaap zoals rapid eye movement (REM) slaap selectief te verstoren. De techniek van zachte behandeling is een krachtig hulpmiddel voor de studie van de effecten van chronische slaap beperking zelfs in de neonatale muizen die veel van de meer stress procedures gebruikt voor slaaptekort omzeilt.

Om te onderzoeken de effecten van chronische slaap beperking van gedrag, slapen we beperkt muizen gedurende drie uur dagelijks tussen 11:00 en 2:00 PM van P5 P428. Naar aanleiding van de beperking van de slaap, waren muizen ongemoeid gelaten voor vier weken en toegestaan om te herstellen van beperking van de slaap. Muizen werden getest voor gedrag na de vier weken durende herstelperiode. De test van sociaal gedrag van de drie-kamer werd gebruikt voor het beoordelen...

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

We beschrijven een slaap deprivatie techniek die bekend staat als zachte behandeling waar onderzoekers zachtjes prod muizen elk moment slaap gedrag wordt waargenomen. Deze methode is een krachtig hulpmiddel waarmee onderzoekers de effecten die chronische slaap beperking in de gehele ontwikkeling op toekomstige hersenen fysiologie en gedrag hebben kan te bestuderen.

Chronisch slaaptekort in muis Pups door middel van zachte behandeling

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

Deze methode kan helpen bij het beantwoorden van belangrijke vragen in het slaapveld, zoals de invloed van chronische slaapbeperking tijdens de postnatale ontwikkeling. Het belangrijkste voordeel van deze techniek is dat het minder stressvol dan andere slaaptekort technieken en kan gemakkelijk worden gebruikt tijdens de ontwikkeling en blijven in de volwassenheid. Begin met het toewijzen van elke kooi aan de slaaptekort of controlegroep.Zorg ervoor dat muizen toegang hebben tot voedsel- en wateradvertentielibitum en verwijder voedsel-, beddengoed of verrijkingsobjecten die het zicht op de muizen kunnen beperken. Voor de controlegroep, verwijder de vader uit de fokker kooi op postnatale dag vijf en laat de dam met de pups tot het spenen. Beweeg vervolgens muizen toegewezen aan de controlegroep naar de slaaptekortkamer en prik de muizen voorzichtig gedurende 10 minuten per dag met een penseel voor de duur van het experiment.Na 10 minuten breng je de muizen terug naar de dierenkamer en verstoor ze niet. Begin met het verwijderen van de vader uit de fokkerskooi op postnatale dag vijf en laat de moeder met de pups tot het spenen. Verplaats muizen toegewezen aan de slaaptekort groep naar de slaaptekort kamer dagelijks tijdens de lichtcyclus waar ze zullen worden gecontroleerd.Gedurende deze tijd, voorzichtig prik de muis met een penseel als vermoed dat in slaap totdat een reactie wordt waargenomen. U muizen omkeren en op hun rug duwen om de slaap te verstoren. Overweeg een muis om te slapen als een van de volgende optreden: trillen, inactiviteit, of als de ogen gesloten zijn.Beschouw muizen wakker te zijn als ze bewegen, proberen om te draaien na op hun rug, of grooming zelf. Als de moeder de pups bedekt tijdens het slaaptekort, prik haar dan voorzichtig weg van de pups, zodat het zicht op de pups onbeperkt is. Ga verder met het slaaptekortprotocol tot het eindpunt van het experiment.Wanneer de periode van drie uur van slaaptekort is voltooid, zet het beddengoed en alle voedsel dat kan zijn verwijderd terug en zet het deksel terug op de kooi. Tot slot, terug kooien naar de dierlijke holding kamer tot de volgende dag van slaaptekort. Na een herstelperiode van een maand gaven de resultaten aan dat in de gezelligheidstest de vrouwtjes in slaap meer tijd besteedden aan het snuiven van de vreemdere muis dan controlevrouwtjes en slaapbeperkte mannetjes de neiging hadden om meer tijd te besteden aan het snuiven van het object in vergelijking met controlemannetjes.Verder, in de sociale nieuwigheid fase van het testen, slaap beperkt vrouwelijke muizen brachten een verhoogde hoeveelheid tijd snuiven de nieuwe muis dan vrouwelijke controles. Tijdens een poging tot deze procedure, is het belangrijk om te onthouden om aandacht te besteden en onmiddellijk te reageren op pre-slaap gedrag, vooral in neonatale muizen waar de slaapbehoefte hoog is. Na deze procedure, andere methoden zoals open veld, rotarod, verhoogde plus doolhof, en sociaal gedrag testen kunnen worden uitgevoerd om aanvullende vragen te beantwoorden, zoals de lange termijn gedragseffecten van chronische slaapbeperking tijdens de ontwikkeling.

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.

De crossmodaal Congruentie Taak als een middel om een ​​doelstelling Behavioral Meet verkrijgen in de 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 ...

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.

Het gebruik van de Puzzle Box als een manier om te beoordelen van de werkzaamheid van Environmental Enrichment

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.

Fouten als een middel om Impulsief Voedsel Keus

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.

Manipulatie van Epileptiforme Electrocorticograms (ECoGs) en Sleep bij ratten en muizen door Acupunctuur

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.

Neurologische Reflex Testing in de neonatale 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.

Noninvasive, High-throughput bepaling van de duur van de slaap in knaagdieren

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.

Meten van actieve en passieve Tameness afzonderlijk in muizen

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.

Roman Objectherkenning en Object locatie gedrags testen in muizen op een begroting

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.

Automatische metingen van slaap en motorische activiteit in Mexicaanse 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.

Een chronisch slaapfragmentatiemodel met trillende orbitale rotor om cognitieve tekorten en angstachtig gedrag op te wekken bij jonge wilde muizen

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