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

Denna forskning har finansierats av intramurala forskningsprogrammet av NIMH (ZIA MH00889). RMS stöddes också av en FRAXA postdoktorsstipendium.

Obs: Alla förfaranden godkändes av det nationella institutet för Mental djur vård och användning kommittén och utförs i enlighet med nationella institut för hälsa riktlinjerna för vård och användning av djur.
1. sleep Deprivation Setup
<ol><li>Använd mjuka penslar för att försiktigt peta möss när du sover.<ol>
<li>Kontrollera att pensel kan passa genom barerna i mus bur mataren så att möss har tillgång till mat och vatten ad libitum under sömnbrist.</li>
		<li>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>

Här beskriver vi en metod för sömnbrist, skonsam hantering, som kan användas i både gnagare ungar och vuxna. Mild lämna kräver forskare att iaktta möss för varaktigheten av en sleep deprivation perioden och försiktigt prod djuren när de är inaktiva eller ryckningar att förhindra sömn. Tidigare sömnstudier frihetsberövande har kontrollerat genom samtidig elektroencefalogram (EEG) inspelningar att varsam hantering förhindrar både REM och icke - REM NREM sömn10. Medan varsam hanter...

Sömnen spelar en avgörande roll i neuronal plasticitet och synaps bildas och eliminering under hjärnans utveckling1,2. Specifikt, är rapid eye movement (REM) sömn viktigt för att bilda stabila synaptic kretsar genom båda stärka specifika synapser och synaptic beskärning2. Farmakologiska sömnbrist tidigt i livet leder till många fysiologiska och beteendemässiga underskott3,4. Förutom farmakologisk sömn deprivation, andra former av sömnbrist som konstant skakningar, tvingade locomotion eller presentation av ett aversivt stimulus har förknippats med depression-liknande symtom under vuxenlivet, olika neurala aktivering mönster och förändringar i sömn tid och sömn kontinuitet under en återhämtningsperiod efter frihetsberövande5,6,7. När sömnen är kroniskt begränsad hos möss i över fem veckor under utveckling, observerades beteendemässiga underskott efter en månad av sömn återvinning8. Sammantaget tyder dessa studier på att störningar av sömn i neonatal gnagare kan påverka senare sömnmönster, hjärnans funktion och beteende. Belysa vikten av sömn för hjärnans normala utveckling hos människor, många patienter med nervsystemets utveckling, inklusive autismspektrumstörningar, tuberös skleros Complex, Fragil X-syndromet och Attention-Deficit/Hyperactivity Sjukdom, rapportera avvikelser i sömnmönster förutom en mängd beteendemässiga underskott1.
Med tanke på antalet kind som är associerade med onormalt sömnmönster, är det viktigt att förstå hur Sömnbrist påverkar hjärnans funktion och beteende. Trots att data som stöder vikten av sömn under utveckling, de flesta metoder som begränsar eller beröva djur av sömn fokuserar på vuxna. Dessa tekniker inkluderar en mängd olika tester som kräver tvingade rörelseaktivitet (t.ex. konstant löpband), konstant störningar (t.ex. roterande sopmaskin bar eller kontinuerlig roman objekt presentation) eller störa djuret vid uppkomsten av REM-sömn (t.ex. plattform över vatten)9,10. Även om dessa metoder har effektivt visat problemen i samband med sömnbrist hos vuxna, är ingen av dessa metoder lämpliga för gnagare valpar på grund av deras begränsade rörlighet, hög sömn drive och de negativa effekterna av stress tidigt i livet. På grund av vikten av sömn under utveckling är det viktigt att förstå hur begränsad sömn i neonatal pups påverkar framtida beteende under vuxenlivet.
En teknik att begränsa sömn som kan användas hos gnagare ungar är varsam hantering. Skonsam hantering innebär att utredaren interagerar direkt med gnagare någon tid sömn beteende observeras. Utredarna kan använda sina händer, en pensel eller presentation av romanen objekt att fysiskt störa sömnen i neonatal gnagare8,10,11,12,13,14 . Sömn kan bestämmas behaviorally av bristande motorisk aktivitet, myokloniska ryckningar och öga stängning (i äldre gnagare pups). EEG validering bekräftar att störa sömnen med varsam hantering baserat på dessa beteenden minskar totala sömntiden med 91% i P12 råttor14. Andra tekniker som har använts för att beröva unga möss inkluderar elstötar och presentation av obehagliga stimuli, men dessa tekniker kan inte användas för kronisk sömn begränsning och är mer stressande än varsam hantering7, 15. kronisk sömn begränsning kan också åstadkommas genom en skakande protokoll som kan användas med eller utan EEG elektrod implantation och tillhörande programvara, men detta protokoll hindrar inte alla sömn och är därför mindre kontrollerade jämfört med varsam hantering4,16. Möjlighet att begränsa eller beröva möss sömn för många dagar i rad utan särskild utrustning, programvara eller elektrod implantation är en fördel av tekniken med varsam hantering. Kronisk sömn begränsning via skonsam hantering effektivt begränsar sömn i neonatal gnagare och producerar en mängd olika beteendeförändringar efter de sleep deprivation periodiska8,11,12, 17.
Här, beskriver vi resultat där kronisk sömnbrist tre timmar per dag från postnatal dag 5 (P5) till P42 avsevärt påverkar sällskaplighet möss efter en 30-dagars återhämtningsperiod där sömn inte störs. Dessa resultat belysa de långsiktiga effekterna av kronisk sömn begränsning under utveckling på framtida beteende.

<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

Sömn är avgörande för god utveckling och neural plasticitet. Dessutom är onormal sömnmönster kännetecknar många neurologiska utvecklingsavvikelser. Studera hur kronisk sömn begränsning under utveckling kan påverka vuxna beteende kan lägga till vår förståelse av uppkomsten av beteendemässiga symtom på kind. Medan det finns många metoder som kan användas för att begränsa sömn hos gnagare inklusive påtvingad förflyttning, ständiga avbrott, presentation av ett aversivt stimulus, eller elektriska stötar, många av dessa metoder är mycket stressande och kan inte användas i neonatala möss. Här beskriver vi skonsam hantering, en sleep deprivation teknik som kan användas för kroniskt hela utveckling och i vuxen ålder för att uppnå sömn begränsning. Skonsam hantering innebär noggrann observation av möss under hela sleep deprivation och kräver forskaren som försiktigt prod djuren när de är inaktiva eller Visa beteenden associerade med sömn. Tillsammans med EEG inspelningar, kunde varsam hantering användas selektivt störa en viss fas av sömn såsom rapid eye movement (REM) sömn. Tekniken med varsam hantering är ett kraftfullt verktyg för att studera effekterna av kronisk sömn begränsning även i neonatala möss som kringgår många av de mer stressande förfaranden som används för sömnbrist.

För att undersöka effekterna av kronisk sömn inskränkning beteende, sover vi begränsad möss i tre timmar dagligen mellan 11:00 och 2:00 PM från P5 till P428. Efter sömn begränsningen, var möss lämnas ostörda i fyra veckor och får återhämta sig från sömn begränsning. Möss testades för beteende efter fyra veckors återhämtningsperiod. Tre-kammare sociala beteende testet användes för att bedöma sällskaplighet (preferens för en roman mus jämf...

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

Vi beskriver en sleep deprivation teknik känd som varsam hantering där utredarna försiktigt prod möss som helst sova beteende observeras. Denna metod är ett kraftfullt verktyg som tillåter forskare att studera effekterna som kronisk sömn begränsning i hela utvecklingen kan få på framtida hjärnans fysiologi och beteende.

Kronisk sömnbrist i musen PUP med hjälp av skonsam hantering

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

Denna metod kan hjälpa till att besvara viktiga frågor i sömnen området såsom påverkar av kronisk sömn begränsning under postnatal utveckling. Den största fördelen med denna teknik är att det är mindre stressande än andra sömnbrist tekniker och kan lätt användas under hela utvecklingen och fortsätta in i vuxen ålder. Börja med att tilldela varje bur till antingen sömnbrist eller kontrollgrupp.Se till att möss har tillgång till mat och vatten ad libitum, och ta bort alla livsmedel, sängkläder eller anrikning objekt som kan begränsa synen på mössen. För kontrollgruppen, ta bort faren från uppfödaren buren på postnatal dag fem och lämna dammen med ungarna tills avvänjning. Därefter flytta möss som tilldelats kontrollgruppen till sömnbristrummet och försiktigt prod mössen med en pensel kontinuerligt i 10 minuter om dagen under hela experimentet.Efter 10 minuter, returnera mössen till djurhållningsrummet och stör dem inte. Börja vid flyttande den sire från uppfödaren bur på postnatal dag fem och lämna den dam med den pups till avvänjningen. Flytta möss som tilldelats gruppen för sömnbrist till sömnbristrummet dagligen under ljuscykeln där de kommer att övervakas.Under denna tid, försiktigt prod musen med en pensel om misstänkt för att sova tills ett svar observeras. Alternativt, invertera möss och tryck över på ryggen för att störa sömnen. Betrakta en mus som sover om något av följande inträffar:ryckningar, inaktivitet eller om ögonen är stängda.Tänk möss att vara vaken om de rör sig, försöker vända efter att ha varit på ryggen, eller grooming själva. Om fördämningen är täckande den pups under sömnen deprivation, försiktigt prod henne bort från pups så synen om pups är unrestricted. Fortsätt med sömnbrist protokoll tills experiment slutpunkten.När tre timmars sömnbrist är klar, lägg sängkläder och all mat som kan ha tagits bort tillbaka och sätta locket tillbaka på buren. Slutligen, returnera burar till djurhållningsrummet till nästa dag av sömnbrist. Efter en månad återhämtningsperiod, visade resultaten att i sällskaplighet testet, sömn begränsade kvinnor tillbringade mer tid sniffa främlingen musen än kontroll honor och sömn begränsade män hade en tendens att spendera mer tid sniffa objektet jämfört med kontroll män.Vidare, i den sociala nyhet fasen av testning, sömn begränsad kvinnliga möss tillbringade en ökad tid sniffa romanen musen än kvinnliga kontroller. Medan du försöker detta förfarande, Det är viktigt att komma ihåg att uppmärksamma och snabbt svara på pre-sömn beteenden, särskilt i neonatal möss där sömn behov är hög. Efter denna procedur, andra metoder som öppet fält, rotarod, förhöjda plus labyrint, och socialt beteende testning kan utföras för att besvara ytterligare frågor som den långsiktiga beteendemässiga påverkar kronisk sömn begränsning under utveckling.

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.

Den Crossmodal kongruens Task som ett sätt att få en objektiv Behavioral Mät i gummihand 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.

Användning av Puzzle Box som ett sätt att bedöma hur effektiva miljöberikning

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.

Fel som ett sätt att minska impulsiv livsmedelsval

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.

Manipulering av Epileptiforma Electrocorticograms (ECoGs) och sömn hos råttor och möss genom akupunktur

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.

Neuro Reflex Testning i neonatala råttungar

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.

Icke-invasiv, High-throughput bestämning av sömn varaktighet hos gnagare

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.

Mätning av aktiv och passiv tamhet separat hos möss

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.

Nya objekt erkännande och objekt plats beteendemässiga testning i möss på en 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.

Automatiska mätningar av sömn och rörelseaktivitet i mexikanska släktet

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.

En kronisk sömnfragmenteringsmodell som använder vibrerande orbitalrotor för att inducera kognitivt underskott och ångestliknande beteende hos unga vilda möss

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