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

Denne forskning er finansieret af murene forskningsprogrammet NiMH (ZIA MH00889). RMS blev også støttet af en FRAXA postdoc stipendium.

Bemærk: Alle procedurer blev godkendt af det nationale Institut for Mental sundhed Animal Care og brug udvalg og udføres i overensstemmelse med de nationale institutter sundhed retningslinjer for pleje og brug af dyr.
1. søvn berøvelse Setup
<ol><li>Brug bløde pensler til at forsigtigt stikke mus når søvn.<ol>
<li>Sørg for pensel kan passe gennem tremmerne af musen bur feeder til at sikre, at mus har adgang til mad og vand ad libitum under søvn.</li>
		<li>Brug ikke pensler...

<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>

Her vi beskrive en metode for søvnmangel, skånsom håndtering, der kan bruges i både gnaver unger og voksne. Blid aflevering kræver forskere at observere mus for varigheden af en søvn berøvelse periode og forsigtigt prod dyrene, når de er inaktive eller trækninger at forhindre søvn. Forudgående søvn berøvelse undersøgelser har bekræftet ved hjælp af samtidige elektroencefalografi (EEG) optagelser, skånsom håndtering forhindrer både REM og ikke - REM NREM søvn10. Mens skånsom h?...

Søvn spiller en afgørende rolle i neuronal plasticitet og synapse dannelse og eliminering under hjerne udvikling1,2. Specifikt, er hurtige øjenbevægelser (REM) søvn afgørende for at danne stabile synaptic kredsløb gennem begge styrkelse af specifikke synapser og synaptic beskæring2. Farmakologiske søvnmangel tidligt i livet fører til mange fysiologiske og adfærdsmæssige underskud3,4. Ud over farmakologiske søvn berøvelse, andre former for søvnmangel som konstant ryster, tvunget bevægelse eller præsentation af et afskrækningsmiddel stimulus har været forbundet med depression-lignende symptomer i voksenalderen, forskellige neurale aktivisering mønstre, og ændringer i søvn tid og søvn kontinuitet i et opsving periode efter afsavn5,6,7. Når søvn er kronisk begrænset i mus i over fem uger under udvikling, blev adfærdsmæssige underskud observeret efter en måned af søvn recovery8. Taget sammen, tyder disse undersøgelser på, at forstyrrelse af søvn i neonatal gnaver kan påvirke senere søvnmønster, hjernefunktion og adfærd. Fremhævelsen af betydningen af søvn i normal hjernens udvikling hos mennesker, mange patienter med nervesystemets, autismespektrumforstyrrelser, knolde sklerose komplekse, skrøbelige X syndrom og Attention-Deficit/Hyperactivity Lidelse, rapport abnormiteter i søvnmønstre ud over en række adfærdsmæssige underskud1.
I betragtning af antallet af nervesystemets, der er forbundet med unormale søvnmønstre, er det vigtigt at forstå, hvordan mangel på søvn påvirker hjernefunktion og adfærd. Trods de data, der understøtter betydningen af søvn under udvikling, de fleste metoder, som begrænser eller fratage dyr i søvn fokus på voksne. Disse teknikker omfatter en række test, der kræver tvunget bevægeapparatet aktivitet (f.eks. konstant løbebånd), konstante forstyrrelser (fx, roterende sweeper bar eller løbende roman objekt præsentation) eller forstyrrende dyr i starten af REM-søvn (fx platform over vand)9,10. Selv om disse metoder har effektivt vist problemerne i forbindelse med søvn hos voksne, er ingen af disse metoder passende for gnavere unger på grund af deres begrænsede mobilitet, høje søvn drev og de negative virkninger af stress tidligt i livet. Betydningen af søvn under udvikling er det vigtigt at forstå, hvordan begrænset søvn i neonatal pups påvirker fremtidige adfærd i voksenalderen.
En teknik til at begrænse søvn, der kan bruges i gnavere unger er skånsom håndtering. Skånsom håndtering indebærer investigator direkte interagere med gnavere enhver tid sleep behavior er observeret. Efterforskere kan bruge deres hænder, en pensel eller præsentation af nye objekter til fysisk forstyrre søvn i neonatal gnavere8,10,11,12,13,14 . Søvn kan bestemmes behaviorally ved manglende motorisk aktivitet, myoklon trækninger og øje lukning (i ældre gnaver pups). EEG validering bekræfter, at forstyrre søvn med skånsom håndtering baseret på disse adfærdsmønstre reducerer total sleep-tid med 91% i P12 rotter14. Andre teknikker, der er blevet brugt til at fratage unge mus omfatter elektrisk stød og præsentation af ubehagelige stimuli, men disse teknikker kan ikke bruges til kronisk søvn begrænsning og er mere stressende end skånsom håndtering7, 15. kronisk søvn begrænsning kan også ske gennem en rystende protokol, der kan bruges med eller uden EEG elektrode implantation og tilknyttede computersoftware, men denne protokol forhindrer ikke alle sove og derfor er mindre kontrolleres i forhold til skånsom håndtering4,16. Evnen til at begrænse eller fratage mus søvn for mange dage i træk uden særlige udstyr, computersoftware eller elektrode implantation er en fordel for skånsom håndtering teknik. Kronisk søvn begrænsning via skånsom håndtering effektivt begrænser søvn i neonatal gnavere og producerer en række adfærdsmæssige ændringer efter den søvn berøvelse periode8,11,12, 17.
Her, beskriver vi resultater som kronisk søvnmangel tre timer om dagen fra postnatal dag 5 (P5) til P42 væsentligt påvirker selskabelighed i mus efter en 30 dages tilbagebetalingsperioden, hvor søvn ikke er forstyrret. Disse resultater fremhæve de langsigtede virkninger af kronisk søvn begrænsning under udvikling på fremtidige adfærd.

<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

Søvn er afgørende for korrekt udvikling og neural plasticitet. Derudover er unormalt søvnmønster karakteristiske for mange forstyrrelser i nervesystemets. At studere hvordan kronisk søvn begrænsning under udvikling kan påvirke voksne adfærd kan føje til vores forståelse af fremkomsten af adfærdsmæssige symptomer på nervesystemets udvikling. Mens der er mange metoder, der kan bruges til at begrænse søvn i gnavere herunder tvungen bevægelse, konstante forstyrrelser, præsentation af et afskrækningsmiddel stimulus, eller elektrisk stød, mange af disse metoder er meget stressende og kan ikke bruges i neonatal mus. Vi beskriver her, skånsom håndtering, en sleep afsavn teknik, der kan bruges kronisk hele udvikling og ind i voksenalderen for at opnå søvn begrænsning. Skånsom håndtering indebærer tæt observation af mus i hele perioden søvn berøvelse og kræver forsker at forsigtigt prod dyrene, når de er inaktive eller vise adfærd forbundet med søvn. Kombineret med EEG optagelser, kan skånsom håndtering anvendes til selektivt forstyrre en bestemt fase i søvn som hurtige øjenbevægelser (REM) søvn. Skånsom håndtering teknik er et kraftfuldt værktøj til undersøgelse af virkningerne af kronisk søvn restriktion selv i neonatal mus, der omgår mange af mere stressende metoder til søvnmangel.

For at undersøge virkningerne af kronisk søvn begrænsning på adfærd, sover vi begrænset mus i tre timer dagligt mellem 11:00:00 og 2:00 fra P5 til P428. Efter søvn begrænsning, var mus venstre uforstyrret i fire uger og lov til at inddrive fra søvn begrænsning. Mus blev testet for adfærd efter fire ugers tilbagebetalingsperioden. De tre-kammer sociale adfærd test blev brugt til at vurdere selskabelighed (præference for en roman mus i forhold til en rom...

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 søvn berøvelse teknik kendt som skånsom håndtering hvor efterforskere forsigtigt prod mus helst sove adfærd er observeret. Denne metode er et kraftfuldt værktøj, der giver forskere til at studere de effekter, som kronisk søvn begrænsning i hele udvikling kan have på fremtidige hjernens fysiologi og adfærd.

Kronisk søvnmangel i mus unger med skånsom håndtering

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

Denne metode kan hjælpe med at besvare centrale spørgsmål i søvn området såsom påvirker af kronisk søvn begrænsning under postnatal udvikling. Den største fordel ved denne teknik er, at det er mindre stressende end andre søvnmangel teknikker og kan nemt bruges i hele udviklingen og fortsætte ind i voksenalderen. Begynd med at tildele hvert bur til enten søvnmangel eller kontrolgruppe.Sørg for, at mus har adgang til mad og vand ad libitum, og fjern alle mad-, strøelses- eller berigelsesobjekter, der kan begrænse musenes udsigt. For kontrolgruppen fjernes faren fra opdrætterburet efter fem dage, og dæmningen efterlades hos ungerne, indtil de fravænner. Dernæst flytte mus tildelt kontrolgruppen til søvnmangel værelse og forsigtigt prod musene med en pensel kontinuerligt i 10 minutter om dagen for varigheden af forsøget.Efter 10 minutter skal musene returneres til dyreholderummet, og de må ikke forstyrres. Begynd med at fjerne faren fra opdrætterburet på postnatal dag fem og lad dæmningen med hvalpene indtil fravænning. Flyt mus tildelt søvnmangel gruppe til søvnmangel værelse dagligt i løbet af lyset cyklus, hvor de vil blive overvåget.I løbet af denne tid, forsigtigt prod musen med en pensel, hvis mistænkt for at være i søvn, indtil en reaktion er observeret. Alternativt, invertere mus og skubbe over på ryggen for at forstyrre søvn. Overvej, at en mus sover, hvis et af følgende forekommer: trækninger, inaktivitet, eller hvis øjnene er lukkede.Overvej mus til at være vågen, hvis de bevæger sig rundt, forsøger at vende efter at være på ryggen, eller grooming sig selv. Hvis dæmningen dækker hvalpene under søvnmangel, forsigtigt prod hende væk fra hvalpene, så udsigten over hvalpene er ubegrænset. Fortsæt med protokollen om søvnmangel, indtil eksperimentslutpunktet.Når de tre timers periode med søvnmangel er færdig, sætte strøelse og mad, der kan have været fjernet tilbage og sætte låget tilbage på buret. Endelig returnere bure til dyret bedrift værelse indtil den næste dag i søvnmangel. Efter en måned restitutionsperiode, viste resultaterne, at i omgængelighed test, søvn begrænset hunner brugt mere tid sniffing den fremmede mus end kontrol hunner og søvn begrænset mænd havde en tendens til at bruge mere tid sniffing objektet i forhold til kontrol mænd.Yderligere, i den sociale nyhed fase af test, søvn begrænset kvindelige mus brugt en øget mængde tid sniffing den nye mus end kvindelige kontrol. Mens du forsøger denne procedure, Er det vigtigt at huske at være opmærksom og straks reagere på pre-sleep adfærd, især i neonatal mus, hvor søvn behov er høj. Efter denne procedure, andre metoder som åbent felt, rotarod, forhøjet plus labyrint, og social adfærd test kan udføres for at besvare yderligere spørgsmål som den langsigtede adfærdsmæssige virkninger af kronisk søvn begrænsning under udvikling.

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 Congruency opgave som middel til at opnå et mål Behavioral måling på 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.

Brugen af ​​Puzzle Box som et middel til vurdering af effektiviteten af ​​miljøberigelse

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.

Fejl som et middel til at reducere Impulsive Food valg

Nowadays, the increasing incidence of eating disorders due to poor self-control has given rise to increased obesity and other chronic weight problems, and ...

Manipulation of Epileptiform Electrocorticograms (ECoGs) and Sleep in Rats and Mice by Acupuncture

Ancient Chinese literature has documented that acupuncture possesses efficient therapeutic effects on epilepsy and insomnia. There is, however, little research to reveal the possible mechanisms behind these effects. To investigate the effect of acupuncture on epilepsy and sleep, several issues need to be addressed. The first is to identify the acupoints, which correspond between humans, rats, and mice. Furthermore, the depth of insertion of the acupuncture needle, the degree of needle twist in manual needle acupuncture, and the stimulation parameters for electroacupuncture (EA) need to be determined. To evaluate the effects of acupuncture on epilepsy and sleep, a feasible model of epilepsy in rodents is required. We administer pilocarpine into the left central nucleus of the amygdala (CeA) to simulate focal temporal lobe epilepsy (TLE) in rats. Intraperitoneal (IP) injection of pilocarpine induces generalized epilepsy and status epilepticus (SE) in rats. Five IP injections of pentylenetetrazol (PTZ) with a one-day interval between each injection successfully induces spontaneous generalized epilepsy in mice. Recordings of electrocorticograms (ECoGs), electromyograms (EMGs), brain temperature, and locomotor activity are used for sleep analysis in rats, while ECoGs, EMGs, and locomotor activity are employed for sleep analysis in mice. ECoG electrodes are implanted into the frontal, parietal, and contralateral occipital cortices, and a thermistor is implanted above the cerebral cortex by stereotactic surgery. EMG electrodes are implanted into the neck muscles, and an infrared detector determines locomotor activity. The criteria for categorizing vigilance stages, including wakefulness, rapid eye movement (REM) sleep, and non-REM (NREM) sleep are based on information from ECoGs, EMGs, brain temperature, and locomotor activity. Detailed classification criteria are stated in the text.

Manipulation of Epileptiform Electrocorticograms ECoGs and Sleep in Rats and Mice by Acupuncture

This paper demonstrates the performance of acupuncture, epilepsy models, and the analysis of sleep in rodents. The acupuncture procedure and the identification of acupoints are described. Pilocarpine or pentylenetetrazol (PTZ) is used to induce epilepsy. Electrocorticogram (ECoG), electromyogram (EMG), brain temperature, and locomotor activity recordings are employed for sleep analysis.

Manipulation af epileptiforme Electrocorticograms (ECoGs) og Sleep i rotter og mus ved 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.

Neurologisk Reflex Afprøvning i Neonatal rotteunger

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.

Noninvasiv, høj overførselshastighed bestemmelse af søvn varigheden i gnavere

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.

Måling af aktive og Passive Tameness separat i mus

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 objekt anerkendelse og objekt placering adfærdsmæssige test i mus 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.

Automatiserede målinger af søvn og bevægeapparatet aktivitet i mexicansk 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.

En kronisk søvnfragmenteringsmodel, der bruger vibrerende orbital rotor til at fremkalde kognitivt underskud og angstlignende adfærd hos unge mus af vildtype

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