Name: the modified hole board - measuring behavior, cognition and social interaction in mice and rats
Uploaded: 2015-04-08T13:00:00
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Description: Watch this Scientific Journal Video about The Modified Hole Board - Measuring Behavior, Cognition and Social Interaction in Mice and Rats at JoVE.com

The authors would like to acknowledge the work of Annemarie Baars and Jos&#233; Lozeman-van &#8216;t Klooster in performing, assisting and teaching experiments using the mHB set-up.

BEMÆRK: Forsøgene er blevet godkendt af Dyreforsøgstilsynet udvalg af University Medical Center Utrecht og Utrecht Universitet, Holland. Desuden dyreforsøg fulgte principperne for laboratoriedyr pleje og henviser til retningslinjerne for pasning og anvendelse af Pattedyr i Neurovidenskab og Behavioural Research. 25 1. forsøgsopstilling BEMÆRK: standard MHB Apparatet består af en grå PVC eksperimentel kasse (100 x 50 x 50 cm) adskilt fra en yderligere rum (50 x 50 x 50 cm), hvor gruppe kammerater i den eksperimentelle dyr kan placeres i testperioden af et gennemsigtigt, perforeret skillevæg 1. Hvis tilstedeværelsen af tilknyttede hjælpere er uønsket, eller hvis individuelt opstaldede dyr testes, udskiftes den gennemsigtige partition af en skillevæg lavet af grå PVC (figur 1;. Se også Ohl et al (2001) 1). Den MHB (with forskellige målinger), der anvendes til kognitiv test er beskrevet i afsnit 5 i protokollen. <ol><li> Placer bord (60 x 20 x 0,5 cm; fremstillet af grå PVC) i midten af ​​kassen. BEMÆRK: Bestyrelsen kan indeholde 20 cylindre (ᴓ 1,5 cm) 14 forskudt i to linjer eller 23 cylindre (ᴓ 3 cm) 8 forskudt i tre linjer. </li><li> Opdel området omkring brættet ved sorte linjer i 10 rektangler (20 x 15 cm) og 2 pladser (20 x 20 cm). </li><li> Placer en scene lys over bord for at skabe en større kontrast i lysintensitet mellem bestyrelsen (der repræsenterer en ubeskyttet område sammenlignes med midten af ​​en åben mark eller det lyse rum af en lys-mørke overgang test) og boksen (beskyttet område) for at øge den afskrækningsmiddel karakter af bestyrelsen 4,8. </li><li> Hus dyrene under en 12 hr omvendt dag-nat cyklus (f.eks lyset slukket på 7:00 AM og lys på 7:00 PM). BEMÆRK: kan imidlertid MHB ogsåanvendes under den konventionelle lys tidsplan (se diskussionen om potentielle mangler) 1,6. </li><li> Udfør den adfærdsmæssige test i den mest aktive fase af dyrene (f.eks, fra 10:00 til 14:00) 4,15. </li><li> Opretholde en tilvænningsperiode på 2 uger efter ankomsten af ​​dyrene i faciliteterne. I denne periode har den samme person, der udfører den adfærdsmæssige eksperiment håndtere dyrene fire gange om ugen og omfatter alle håndteringsprocedurer, der skal udføres under selve prøvningen. Håndter dyrene udelukkende i den tid af dagen, hvor dyrene vil blive udsat for forsøg senere. </li><li> Optag forsøgene til datalagring på video og for at optimere resultaterne og reducere inter variabilitet observatør, øve adfærdsmæssige scoring ved hjælp af en videooptagelse af tidligere forsøg 2. Sørg for at standardisere alle handlinger og procedurer udført af observatør. </li></ol> 2. Behavioral Testing - uden tilstedeværelse af koncernens Mates <ol><li> Udfør adfærdsmæssige test i rummet dyrene regelmæssigt anbragt i (for at undgå mulige virkninger af transport til en test placering) og installere alle testudstyr før ankomst af dyrene i de faciliteter (at vænne dyrene til tilstedeværelsen af ​​udstyret). </li><li> Saml dyret ved haleroden fra sit hjem bur og direkte placere den i MHB. </li><li> Placer hvert dyr i apparatet i det samme hjørne mod væggen (som angivet i figur 1). </li><li> Lad dyret frit udforske MHB for en periode (ofte 5 min 1,6,16-18). </li><li> Har du en erfaren observatør levende score de adfærdsmæssige parametre ved hjælp adfærdsmæssige scoring software. Brug parametrene i tabel 1. BEMÆRK: Visse adfærdsmæssige parametre (fx motor- og udforskende adfærd) kunne scoret automatisk som fx, </em&gt; Diskuteret af Henry et al. (2010) 19 efter nødvendig tilpasning af MHB. </li><li> Rengør apparat med ledningsvand og en papirserviet efter hver prøve for at undgå en bias baseret på olfaktoriske signaler. BEMÆRK: Mulige test ordens effekter Ved test huses socialt dyr skal holdes for øje 17,20. </li></ol> 3. Behavioral Testing - i overværelse af Group Mates <ol><li> I tilfælde af gruppe boliger, måle interaktionen med forsøgsdyret og dens burkammerater under prøvningen. <ol><li> Placer gruppe kammerater i gruppen rum før afprøvning af forsøgsdyr for at tillade tilvænning (hovedsagelig 10-30 min 1,12). BEMÆRK: Test under social-stressbetingelser er muligt ved at placere en dominerende bur-mate i gruppen rummet ved test af en socialt besejret individuel 13. </li><li> Placer forsøgsdyret i testen rum og gøre det muligt at frigørely udforske MHB som beskrevet i afsnit 2. BEMÆRK: Mulige test ordens effekter Ved test huses socialt dyr skal holdes for øje 17,20. </li><li> Har du en erfaren observatør levende score de adfærdsmæssige parametre ved hjælp adfærdsmæssige scoring software. Brug parametrene i tabel 1. </li><li> Rengør apparat med ledningsvand og en papirserviet efter hver prøve for at undgå en bias baseret på olfaktoriske signaler. </li></ol></li></ol> 4. Novel Object Anerkendelse og fødeindtag Inhibition <ol><li> Gør dyrene med en genstand (for eksempel en terning eller en fødevare pellet) i deres hjem bur 2 dage før eksperimentet. </li><li> Placer bekendt objekt i apparatet 2 cm bortset fra en hidtil ukendt genstand (for eksempel en bolt eller ukendte mad) i hjørnet på tværs fra startpunktet. </li><li> Den tid dyret tager at nærme romanen og velkendte objekt / mad. Brug parametre &lt;strong&gt; Tabel 1. </li><li> Rengør apparat med ledningsvand og en papirserviet efter hver prøve for at undgå en bias baseret på olfaktoriske signaler. </li></ol> 5. Kognitiv Test <ol><li> Placer en mindre bord (35 x 22 x 1 cm) med 10 cylindre i midten af kassen (figur 1) til afprøvning rotter 3,22. Reducer boksen i størrelse til 50 x 50 cm til test mus 21 ved at indsætte en skillevæg lavet af grå PVC. </li><li> Duft alle cylindre med en smag dyr er tiltrukket af (f.eks vanille) og agn alle med en belønning (fx et stykke mandel, en meget velsmagende belønning for mus og rotter) under et gitter, så dyrene ikke kan fjerne det. </li><li> Cue cylindre (ofte tre) med en farvet ring (kontrast til den grå PVC) og agn dem med en aftagelig belønning (fx 0,05 g stykke mandel). </li><li> Gør dyrene med belønning dagligt i 2 dage før forsøget ideres hjem bur ved at tilbyde det med en pincet og sikre, at dyrene spiser den. </li><li> Har du en erfaren observatør levende score de adfærdsmæssige parametre ved hjælp adfærdsmæssige scoring software. Mål parametrene i tabel 2 i tillæg til de adfærdsmæssige parametre, der er nævnt i § 2 (tabel 1) med undtagelse af parametrene vedrørende indsigelse anerkendelse eller fødeindtagelse hæmning. </li><li> Trin 1: Med hvert dyr, udføre fire forsøg dagligt med en konstant inter-trial interval (fx 30-60 min), indtil en konstant tid til at afslutte en retssag er nået (dvs. når alle tre mandel stykker er blevet indsamlet). </li><li> Trin 2: Cue og agn tre forskellige cylindre og placere dyrene i opsætningen for fire forsøg for at teste tilbageførslen indlæringsevnen. </li><li> Rengør apparat med ledningsvand og en papirserviet efter hver prøve for at undgå en bias baseret på olfaktoriske signaler. </li></ol>

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The authors have nothing to disclose.

Den MHB paradigme kan anvendes til at måle flere dimensioner konditioneret adfærd. Protokollen kan ændres en smule, afhængigt af formålet med forsøget. I denne protokol vi diskutere indstillingerne tider og målinger normalt bruges i vores laboratorium. Imidlertid har små afvigelser til målingerne af apparatet blevet brugt i fortiden og også mængden af flasker i bestyrelsen kan variere 3. Ofte undersøgelser ansætte en testtid på 5 min pr forsøg, men andre test tider kan også være hensigtsmæssig, dvs. sluttede testen, så snart en kognitiv forsøg er blevet gennemført med succes eller udvide test tid, hvis dyrene er ekstremt nervøse eller fysisk svækket. Tidspunktet på dagen for testning blev valgt til at være under rødt lys tilstand, da gnavere er nataktive dyr, og er mest aktive i den tidlige mørke fase. Roedel et al. (2006) viser virkningerne af lys eller mørk fase test på adfærdsmæssige og kognitive præstationeri DBA mus i MHB 16. Andre undersøgelser har udført MHB eksperimenter under hvide lysforhold 1,6, men det skal bemærkes, at teste under hvidt lys betingelser kan fremkalde adfærdsmæssige hæmning og kognitive forstyrrelser (som vist i DBA mus) 16. Tabel 1 og 2 indeholder en stor mængde af adfærdsmæssige parametre, der skal måles. Under dataanalyse kan det føre til nogle parametre indikerer en betydelig stigning på eksempelvis &quot;latens til første indrejse bord &#39;, men ikke i andre parametre af samme motiverende systemet (i dette tilfælde&quot; Undgåelse &quot;). I nogle tilfælde kan det føre til overbevisende resultater. Guilloux et al. (2011) indført integrerede adfærdsmæssige z-scores til den adfærdsmæssige fænotypebestemmelse i mus 24. Med brugen af ​​integrerede adfærdsmæssige z-scorer flere parametre kan kombineres til en enkelt z-score beskriver en særlig motivational system. De efterfølgende z-scores kan på deres side være lettere sammenlignes på tværs af adfærdsmæssige forsøg og eksperimenter. Ud over de beskrevne træk i dette paradigme, en mere dybtgående brug blev klart i undersøgelsen af Salomons et al. (2012). Den tilvænning til nyhed af to musestammer (Balc / cJ og 129P3 / J) i MHB blev sammenlignet, udviser en forskel i adfærdsmæssige fleksibilitet indikerer en ikke-adaptiv adfærdsmæssige profil 129P3 / J-mus 4, spejling forringet adaptive kapacitet og sandsynligvis selv patologisk angst. Afsluttende, den MHB tillader opmåling af flere adfærdsmæssige dimensioner i et enkelt forsøg. Ved at kombinere funktioner fra en traditionel hul bord og åben mark test, ubetinget adfærd, social interaktion, kognition og adaptive kapacitet, dvs. kan velfærd undersøges. Denne test kan for eksempel anvendes til vurdering af adfærdsmæssige ændringer som følge pharmacological- og / ellergenetiske manipulationer, selektiv avl og adaptive kapacitet. I forhold til klassiske test batterier, er antallet af dyr, der er nødvendige reducerede tydeligt og stress opleves af dyrene under test er ekstremt lav.

Den modificerede hul board (MHB) bruges til at vurdere flere dimensioner af ubetinget adfærd, primært i mus og rotter 1. En række almindeligt anvendte tests måle en enkelt adfærdsmæssige parameter, som ikke fuldstændigt dækker hele fænotype af en adfærdsmæssig dimension. Den MHB er udviklet ud fra det koncept, at gnavere kan vise deres rige adfærdsrepertoire kun i et rigt testmiljø 2 og dermed giver mulighed for komplekse etologiske observationer. Det set-up omfatter karakteristika traditionelle hul bord og åben mark test, hvilket resulterer i et enkelt kompleks paradigme, som overvinder ulemperne ved en test batteri (dvs. at reducere antallet af dyr, der anvendes 1,3,4, omgå mulige virkninger af test orden 5, og reducere time-effekt og omkostninger 6). I modsætning til de fleste adfærdsmæssige tests (f.eks Hanell &amp; Marklund, 2014) 7 &lt;/ Sup&gt;, en fordel ved MHB er, at dyr ikke behøver at være mad berøvet for at øge motivationen for at løse opgaven. Derudover kan social isolation omgås under test ved at placere gruppe kammerater i forsøgsordningen dyr i en (gruppe-) rum adskilt fra test rummet ved en gennemsigtig perforeret skillevæg, der giver mulighed for visuel, auditiv og olfaktoriske kontakt 8,9. Den MHB er (farmakologisk) valideret for både mus og rotter 1,6. Kan måles en bred vifte af adfærd, såsom undgåelsesadfærd, risikovurdering, ophidselse, udforskning, lokomotorisk aktivitet, tilvænning, social affinitet og kognition 2,8-10. Derudover kan MHB kombineres med en fødeindtagelse inhibition test, samt nye objekt anerkendelse test 10,11. Endelig kan MHB også bruges til at udføre sociale stress eksperimenter ved at teste socialt besejrede dyr mens placere en dominerende person igruppen rummet 12,13. Denne protokol for mus og rotter vil give et overblik over de mange anvendelser af MHB.

<table border="0" cellpadding="0" cellspacing="0" width="843">
	<tbody>
		<tr height="42">
			<td height="42" style="height:42px;width:211px;">Name of Reagent/ Equipment</td>
			<td style="width:133px;">Company</td>
			<td style="width:119px;">Catalog Number</td>
			<td style="width:380px;">Comments/Description</td>
		</tr>
		<tr height="254">
			<td height="254" style="height:255px;width:211px;">Standard mHB apparatus</td>
			<td style="width:133px;">N/A</td>
			<td style="width:119px;">N/A</td>
			<td style="width:380px;">The standard mHB apparatus consists of a grey PVC experimental box (100x50x50 cm) separated from a group compartment (50x50x50 cm) by a transparent, perforated partition. If the presence of group mates is unwanted or if individually housed animals are tested, the transparent partition is replaced by a partition made of grey PVC. The board (60x20x0.5 cm; made of grey PVC) is placed in the middle of the box and can contain 20 cylinders (&#7443; 1.5 cm) staggered in two lines or 23 cylinders (&#7443; 3 cm) staggered in three lines positioned on the board. The area around the board is divided by black lines into 10 rectangles (20x15 cm) and 2 squares (20x20 cm).</td>
		</tr>
		<tr height="210">
			<td height="210" style="height:210px;width:211px;">Cognitive mHB apparatus</td>
			<td style="width:133px;">N/A</td>
			<td style="width:119px;">N/A</td>
			<td>The box for cognitive testing is 50x50 cm by inserting a partition made of grey PVC in the standard mHB. A smaller board (35x22x1 cm) with 10 cylinders is placed in the middle of the box. All cylinders are scented with a flavor animals are attracted to (e.g. vanilla) and all are baited with a reward (e.g. a piece of almond) beneath a grid so the animals cannot remove it. Cylinders (often three) are cued with a colored ring (contrasting with the grey PVC) and are baited with a removable reward (e.g. 0.05 g piece of almond).&#160;</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:211px;">Vanilla flavor</td>
			<td style="width:133px;">N/A</td>
			<td style="width:119px;">N/A</td>
			<td>Vanilla flavor dissolved in water (0.02%)</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:211px;">Camera</td>
			<td style="width:133px;">N/A</td>
			<td style="width:119px;">N/A</td>
			<td>Camera that generates sufficient quality output to rescore the behavior from video.&#160;</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:211px;">Stage light</td>
			<td style="width:133px;">N/A</td>
			<td style="width:119px;">N/A</td>
			<td>A theather light that allows to adjust the light intensity and the dimensions of the surface that is lit.&#160;</td>
		</tr>
		<tr height="88">
			<td height="88" style="height:88px;width:211px;">Behavioral scoring software</td>
			<td style="width:133px;">N/A</td>
			<td style="width:119px;">N/A</td>
			<td>Behavioral scoring software.</td>
		</tr>
	</tbody>
</table>

the modified hole board - measuring behavior, cognition and social interaction in mice and rats

Denne protokol beskriver den modificerede hul bord (MHB), som kombinerer funktioner fra en traditionel hul bord og åben mark, og er designet til at måle flere dimensioner af ubetinget adfærd i små laboratoriepattedyr (fx mus, rotter, træspidsmus og små primater). Dette paradigme er et værdifuldt alternativ til anvendelsen af ​​en adfærdsmæssig testbatteri, eftersom et bredt adfærdsmæssige spektrum af et dyrs adfærdsmæssige profil kan undersøges i en enkelt test. Apparatet består af en kasse, der repræsenterer den »beskyttet« område, adskilt fra en gruppe rum. Et bord, hvor små cylindre er forskudt i tre linjer, er placeret i midten af ​​kassen, der repræsenterer &#39;ubeskyttede&#39; område af set-up. De kognitive evner af dyr kan måles ved baiting nogle cylindre på tavlen og måle arbejdsmiljøet og reference hukommelse. Andre ubetinget adfærd, såsom Aktivitety-related-, angst-related- og social adfærd, kan observeres ved hjælp af denne paradigme. Behavioral fleksibilitet og evne til at vænne et hidtil ukendt miljø kan desuden observeres ved at udsætte dyrene for flere forsøg i MHB, afslørende indblik i dyrenes adaptive kapacitet. På grund af at teste ordens virkninger i en adfærdsmæssig test batteri, bør naive dyr skal bruges til hvert enkelt eksperiment. Ved at teste flere adfærdsmæssige dimensioner i et enkelt paradigme og dermed omgå dette problem er antallet af forsøgsdyr, der anvendes reduceret. Endvidere, ved at undgå social isolation under test og uden behov for fødevarer fratage dyrene, MHB repræsenterer en adfærdsmæssig testsystem, inducere eventuelle meget lav mængde af stress.

Den store mængde af parametre, der kan måles i MHB gøre dette setup særlig egnet til at måle talrige adfærdsmæssige dimensioner. Et eksempel er identifikationen af ​​adfærdsmæssige tilpasning til en ny miljø ved gentagen eksponering på prøve. . Salomons et al (2010) undersøgte tilvænning af to indavlede stammer af mus (BALB / cJ og 129P3 / J) til MHB under to forskellige lysforhold (rødt lys: kontrasten mellem kasse og bord: 45 lux vs hvidt lys: kontrast mellem boksen og pension: 115 lux (se også protokol afsnit 1.3)) 4. Balc / cJ mus viser en faldende (habituating) latenstid indtil den første bestyrelse post (se tabel 1) under rødt lys, som vist i figur 2A. I modsætning hertil 129P3 / J-mus viser ingen tegn på tilvænning i forsøg. Figur 2B viser eksperimentet under hvide lysforhold. BALB / cJ mus viser igen en faldende ventetid til den første bestyrelse er trådt i forsøg omend at ennimals viser en langsommere tilvænning mønster i forhold til det røde lys. 129P3 / J-mus ikke kun vise igen svækket tilvænning, men også en tendens til overfølsomhed under den hvide lys. Tilsvarende i en undersøgelse foretaget af Salomons (2012) 129P2 / OlaHsd mus viste en forringet adfærdsmæssige fleksibilitet som reaktion på nyhed i forhold til BALB / cOlaHsd mus 23. Forskellen i tilvænning evne bliver således tydeligt mellem to indavlede stammer af mus, når de testes i MHB 4. Den kognitive version af MHB kan for eksempel anvendes til at måle kognitive svækkelser hos mus. Van der Kooij et al. (2010) brugt denne opsætning til at måle den kognitive funktion C57BL / 6J mus med mild cerebral hypoxi-iskæmi (45 min af hypoxi, HI-45), alvorlig HI (75 min af hypoxi, HI-75) og forsøg med simuleret injektionskontrol kontrolmus 9. Evnen til at gennemføre forsøgene (dvs. finde madding huller inden for 5 min) er vist i figur 3A. </strong&gt; Antallet af korttidshukommelsen fejl (genbesøger til en sat agn hul), langtidshukommelse fejl (besøg på en ikke-agn hul) og undladelse fejl (ingen besøg på en sat agn hul) er vist i figur 3B-D henholdsvis. For at bekræfte, at HI-45 gruppe havde ingen kognitiv svækkelse, blev denne gruppe testet mod de sham-kontroller i en tilbageførsel opgave. De tre madding huller blev udnævnt til tre forskellige cylindre og dyrene blev testet i 4 forsøg. Tilbageførslen effekt viser sig, når man sammenligner det sidste forsøg af første etape med den første retssag af tilbageførslen fase. Varigheden at fuldføre fire vending forsøg giver en indikation på den samlede præstation. Figur 4 viser latenstiden for at afslutte retssagen for både grupper og en klar overordnet behandlingseffekt er indlysende. Det betyder, at tilbageførslen opgave der faktisk en forringelse i kognitiv fleksibilitet (re-learning) i HI-45 gruppe, der er<html> kom påviselig ved hjælp af MHB 9. <img alt="Figur 1" src="/files/ftp_upload/52529/52529fig1.jpg" /> Figur 1. Skematisk oversigt over den modificerede hul bord. (A) set-up består af en test rum (boks) med i midten af (ubeskyttet) bord angivet med zigzag linjer og gruppen rum. <img alt="Figur 5" src="/files/ftp_upload/52529/52529fig5.jpg" /> = Roman objekt, <img alt="Figur 6" src="/files/ftp_upload/52529/52529fig6.jpg" /> = Velkendt objekt, <img alt="Figur 7" src="/files/ftp_upload/52529/52529fig7.jpg" /> = (C) fra siden af en cylinder udgangspunkt. (B) Skematisk oversigt over den kognitive version af den modificerede hul bord for mus. Som anvendt i den kognitive version af den modificerede hul bord. Et stykke af mandel anbringes under gitteret af alle cylindrene.target = &quot;_ blank&quot;&gt; Klik her for at se en større udgave af dette tal. <img alt="Figur 2" src="/files/ftp_upload/52529/52529fig2.jpg" /> Figur 2. Rødt lys eller hvide lysforhold. (A) Latency indtil første board indgang (middel + SEM) af BALB / cJ og 123P3 / J-mus målt under rødt lys (rubrik 0-5 lux og bestyrelse 45 lux). En gentagne målinger ANOVA med Huyn-Feldt justering afslørede en stamme (P = 0,025), forsøg (p &lt;0,001) og retssagen x stamme interaktion (P &lt;0,001) effekt Indlæg hoc analyser:. Mellem stammer: * = p &lt;0,0026, mellem to på hinanden følgende forsøg: $ = P &lt;0,0026 (BALB / cJ) (B) latenstiden for den første bestyrelse indrejse (middel + SEM) under hvide lysforhold (rubrik 0-5 lux og bestyrelse 120 lux).. En gentagne målinger ANOVA med Huyn-Feldt justering afslørede en stamme (P = 0,031), forsøg (p &lt;0,001) og retssagen x stamme interaktion (P &lt;0,001) effe. ct Indlæg hoc analyser: mellem stammer: * = p &lt;0,0026, mellem to på hinanden følgende forsøg: $ = P &lt;0,0026 (BALB / cJ) og T = P &lt;0,0026 (123P3 / J). Dette tal er blevet ændret fra 2010 4 Salomons et al.. <a href="https://www.jove.com/files/ftp_upload/52529/52529fig2highres.jpg" target="_blank">Klik her for at se en større udgave af dette tal.</a> <img alt="Figur 3" src="/files/ftp_upload/52529/52529fig3.jpg" /> Figur 3. Kognitiv test i MHB (fase 1). Alle tal viser middelværdi + SEM. (A) latenstid til at fuldføre forsøget (sek) i skin-kontrolmus, HI-45 og HI-75. (B) Antal undladelse fejl, (C) Ikke-madding hul besøg og (D) genbesøger for agn huller. * = P &lt;0,05, ** = P &lt;0,01, *** = p &lt;0,001 sham vs. HI-75, # = P &lt;0,05, ## = P &lt;0,01, ### = P &lt;0,001HI-45 vs. HI-75. Dette tal er blevet ændret fra Van der Kooij et al. 2010 9. <a href="https://www.jove.com/files/ftp_upload/52529/52529fig3highres.jpg" target="_blank">Klik her for at se en større udgave af dette tal.</a> <img alt="Figur 4" src="/files/ftp_upload/52529/52529fig4.jpg" /> Figur 4. Tilbageførsel learning opgave (trin 2). Latency at fuldføre en retssag (sek) i tilbageførslen learning opgave (middel + SEM). Trial effekter: # = P &lt;0,05, ## = P &lt;0,01 (første retssag tilbageførsel opgave vs. tidligere forsøg). Behandling effekt: * = P &lt;0,05, ** = P &lt;0,01 (HI-45 vs. sham). Dette tal er blevet ændret fra Van der Kooij et al. 2010 9. <table border="0" cellpadding="0" cellspacing="0"><tbody><tr><td> System </td><td> Aktivitet </td><td> Målt parameter </td></tr><tr><td> AvoiDance </td><td> Board post </td><td> Frekvens, latency (r), varighed (%) og gennemsnitlig varighed (r) i bestyrelsen </td></tr><tr><td> Risikovurdering </td><td> Strakte Attends </td><td> Frekvens og latency (r) strakte kropsstillinger (herunder bagbenene) </td></tr><tr><td> Arousal </td><td> Grooming </td><td> Frekvens, latency (r), varighed (%) og gennemsnitlig varighed (s) selv-grooming </td></tr><tr><td></td><td> Defækation </td><td> Frekvens og latency (r) boli produceret </td></tr><tr><td></td><td> Vandladning </td><td> Frekvens og latency (r) urinations </td></tr><tr><td> Instrueret udforskning </td><td> Hole besøg </td><td> Frekvens og latency (r) cylinder besøg </td></tr><tr><td></td><td> Novel objekt udforskning </td><td> Frekvens, latency (r), varighed (%) og gennemsnitlig varighed (s) udforske nye objekt </td></tr><tr><td> Ikke-orienteret udforskning </td><td> Opdræt box &lt;/ Td&gt; <td> Frekvens og latency (r) på bagbenene i kassen (forpoter ikke røre væggen) </td></tr><tr><td></td><td> Opdræt bord </td><td> Frekvens og latency (r) på bagbenene på tavlen </td></tr><tr><td></td><td> Hul udforskning </td><td> Frekvens og latency (r) cylinder udforskninger </td></tr><tr><td> Hukommelse </td><td> Velkendt objekt udforskning </td><td> Frekvens, latency (r), varighed (%) og gennemsnitlig varighed (s) udforske velkendte objekt </td></tr><tr><td> Social affinitet </td><td> Interaktion Group </td><td> Frekvens, latency (r), varighed (%) og gennemsnitlig varighed (r) interagerer med gruppen rum </td></tr><tr><td> Lokomotorisk aktivitet </td><td> Linie passage </td><td> Frekvens og latency (r) linjekrydsninger </td></tr></tbody></table> Tabel 1: Liste over adfærdsmæssige parametre <table border="0" cellpadding="0" cellspacING = &quot;0&quot;&gt; <tbody><tr><td> Memory-system </td><td> Parameter </td><td> Beskrivelse </td></tr><tr><td> Langtidshukommelse </td><td> Forkert valg </td><td> Besøg til ikke-agn cylinder; næse under kanten </td></tr><tr><td> Udeladelse fejl </td><td> Udeladelse af en agn cylinder </td><td> Intet besøg i en agn cylinder </td></tr><tr><td> Korttidshukommelsen </td><td> Gentagen valg </td><td> Vende til agn cylinder; næse under kanten </td></tr><tr><td> Samlet ydelse </td><td> Samlet retssag tid </td><td> Tid indtil alle agn cylindre er blevet besøgt </td></tr></tbody></table> Tabel 2: Liste over kognitive parametre

Watch this Scientific Journal Video about The Modified Hole Board - Measuring Behavior, Cognition and Social Interaction in Mice and Rats at JoVE.com

The Modified Hole Board - Measuring Behavior, Cognition and Social Interaction in Mice and Rats

This protocol describes the modified hole board, which is a behavioral test set-up that comprises the characteristics of an open field and a traditional hole board. This set-up enables the differential analysis of unconditioned behavior of small laboratory mammals as well as the analysis of cognitive abilities.

Den modificerede Hole Board - Måling Behavior, Erkendelse og social interaktion hos mus og rotter

This protocol describes the modified hole board (mHB), which combines features from a traditional hole board and open field and is designed to measure ...

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Research

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Behavior

The Successive Alleys Test of Anxiety in Mice and Rats

The plus-maze was derived from the early work of Montgomery. He observed that rats tended to avoid the open arms of a maze, preferring the enclosed ones. Handley, Mithani and File et al. performed the first studies on the plus-maze design we use today, and in 1987 Lister published a design for use with mice. 
Time spent on, and entries into, the open arms are an index of anxiety; the lower these indices, the more anxious the mouse is. Alternatively, a mouse that spends most of its time in the closed arms is classed as anxious. 
One of the problems of the plus-maze is that, while time spent on, and entries into, the open arms is a fairly unambiguous measure of anxiety, time in the central area is more difficult to interpret, although time spent here has been classified as &#x201C;decision making&#x201D;. In many tests central area time is a considerable part of the total test time.
Shepherd et al. produced an ingenious design to eliminate the central area, which they called the &#x201C;zero maze&#x201D;. However, although used by several groups, it has never been as widely adopted as the plus-maze. 
In the present article I describe a modification of the plus-maze design that not only eliminates the central area but also incorporates elements from other anxiety tests, such as the light-dark box and emergence tests. It is a linear series of four alleys, each having increasing anxiogenic properties. It has given similar results to the plus-maze in general. Although it may not be more sensitive than the plus-maze (more data is needed before a firm conclusion can be reached on this point), it provides a useful confirmation of plus-maze results which would be useful when, for example, only a single example of a mutant mouse was available, as, for example, in ENU-based mutagenesis programs.

The plus-maze measures anxiety-like behaviour in rodents. There are two opposite closed and two opposite open arms; anxious rodents avoid the open arms. The central area is neither completely open nor closed, so time spent here is ambiguous and difficult to interpret. Here a modification of the plus-maze protocol eliminating this area is described.

Den Successiv Alleys Test af angst hos mus og rotter

The plus-maze was derived from the early work of Montgomery. He observed that rats tended to avoid the open arms of a maze, preferring the enclosed ones.  ...

Moderate Prenatal Alcohol Exposure and Quantification of Social Behavior in Adult Rats

Alterations in social behavior are among the major negative consequences observed in children with Fetal Alcohol Spectrum Disorders (FASDs). Several independent laboratories have demonstrated robust alterations in the social behavior of rodents exposed to alcohol during brain development across a wide range of exposure durations, timing, doses, and ages at the time of behavioral quantification. Prior work from this laboratory has identified reliable alterations in specific forms of social interaction following moderate prenatal alcohol exposure (PAE) in the rat that persist well into adulthood, including increased wrestling and decreased investigation. These behavioral alterations have been useful in identifying neural circuits altered by moderate PAE1, and may hold importance for progressing toward a more complete understanding of the neural bases of PAE-related alterations in social behavior. This paper describes procedures for performing moderate PAE in which rat dams voluntarily consume ethanol or saccharin (control) throughout gestation, and measurement of social behaviors in adult offspring.

The goal of the protocol presented here is to describe procedures to expose rats to moderate levels of alcohol during prenatal brain development and to quantify resulting alterations in social behavior during adulthood.

Moderat Prænatal Alkohol Eksponering og kvantificering af social adfærd hos voksne rotter

Alterations in social behavior are among the major negative consequences observed in children with Fetal Alcohol Spectrum Disorders (FASDs). Several ...

Assessment of Social Cognition in Non-human Primates Using a Network of Computerized Automated Learning Device (ALDM) Test Systems

Fagot &#38; Paleressompoulle1 and Fagot &#38; Bonte2 have published an automated learning device (ALDM) for the study of cognitive abilities of monkeys maintained in semi-free ranging conditions. Data accumulated during the last five years have consistently demonstrated the efficiency of this protocol to investigate individual/physical cognition in monkeys, and have further shown that this procedure reduces stress level during animal testing3. This paper demonstrates that networks of ALDM can also be used to investigate different facets of social cognition and in-group expressed behaviors in monkeys, and describes three illustrative protocols developed for that purpose. The first study demonstrates how ethological assessments of social behavior and computerized assessments of cognitive performance could be integrated to investigate the effects of socially exhibited moods on the cognitive performance of individuals. The second study shows that batteries of ALDM running in parallel can provide unique information on the influence of the presence of others on task performance. Finally, the last study shows that networks of ALDM test units can also be used to study issues related to social transmission and cultural evolution. Combined together, these three studies demonstrate clearly that ALDM testing is a highly promising experimental tool for bridging the gap in the animal literature between research on individual cognition and research on social cognition.

Assessment of Social Cognition in Non-human Primates Using a Network of Computerized Automated Learning Device ALDM Test Systems

Fagot &#38; Paleressompoulle1 have published an automated learning device (ALDM) aimed at testing individual cognitive abilities in semi-free ranging monkeys. The main goal of our protocol is to use a network of ALDM test units to study social cognition in non-human primates.

Vurdering af Social Cognition i ikke-menneskelige primater Ved hjælp af en netværk af edb Automated Learning Device (ALDM) Test Systems

Fagot &#38; Paleressompoulle1 and Fagot &#38; Bonte2 have published an automated learning device (ALDM) for the study of cognitive abilities of monkeys ...

The Social Dimension of Stress: Experimental Manipulations of Social Support and Social Identity in the Trier Social Stress Test

In many situations humans are influenced by the behavior of other people and their relationships with them. For example, in stressful situations supportive behavior of other people as well as positive social relationships can act as powerful resources to cope with stress. In order to study the interplay between these variables, this protocol describes two effective experimental manipulations of social relationships and supportive behavior in the laboratory. In the present article, these two manipulations are implemented in the Trier Social Stress Test (TSST)&#8212;a standard stress induction paradigm in which participants are subjected to a simulated job interview. More precisely, we propose (a) a manipulation of the relationship between different protagonists in the TSST by making a shared social identity salient and (b) a manipulation of the behavior of the TSST-selection committee, which acts either supportively or unsupportively. These two experimental manipulations are designed in a modular fashion and can be applied independently of each other but can also be combined. Moreover, these two manipulations can also be integrated into other stress protocols and into other standardized social interactions such as trust games, negotiation tasks, or other group tasks.

Previous research on the social dimension of stress has focused on two important variables: social identity and social support. This protocol introduces an effective experimental manipulation of these two social variables and describes their implementation in a standard stress induction paradigm (Trier Social Stress Test).

Den sociale dimension af stress: Eksperimentelle manipulationer af social støtte og social identitet i Trier Social stresstesten

In many situations humans are influenced by the behavior of other people and their relationships with them. For example, in stressful situations ...

A Method for Evaluating the Reinforcing Properties of Ethanol in Rats without Water Deprivation, Saccharin Fading or Extended Access Training

Operant oral self-administration methods are commonly used to study the reinforcing properties of ethanol in animals. However, the standard methods require saccharin/sucrose fading, water deprivation and/or extended training to initiate operant responding in rats. This paper describes a novel and efficient method to quickly initiate operant responding for ethanol that is convenient for experimenters and does not require water deprivation or saccharin/sucrose fading, thus eliminating the potential confound of using sweeteners in ethanol operant self-administration studies. With this method, Wistar rats typically acquire and maintain self-administration of a 20% ethanol solution in less than two weeks of training. Furthermore, blood ethanol concentrations and rewards are positively correlated for a 30 min self-administration session. Moreover, naltrexone, an FDA-approved medication for alcohol dependence that has been shown to suppress ethanol self-administration in rodents, dose-dependently decreases alcohol intake and motivation to consume alcohol for rats self-administering 20% ethanol, thus validating the use of this new method to study the reinforcing properties of alcohol in rats.

This protocol describes a novel and efficient method to quickly initiate operant responding for ethanol in rats that, contrary to standard methods, does not require water deprivation or saccharin/sucrose fading to initiate responding.

En metode til evaluering af forstærkende egenskaber af ethanol i rotter uden vand afsavn, Saccharin Fading eller udvidet adgang Uddannelse

Operant oral self-administration methods are commonly used to study the reinforcing properties of ethanol in animals. However, the standard methods ...

Examining Recall Memory in Infancy and Early Childhood Using the Elicited Imitation Paradigm

The ability to recall the past allows us to report on details of previous experiences, from the everyday to the significant. Because recall memory is commonly assessed using verbal report paradigms in adults, studying the development of this ability in preverbal infants and children proved challenging. Over the past 30 years, researchers have developed a non-verbal means of assessing recall memory known as the elicited or deferred imitation paradigm. In one variant of the procedure, participants are presented with novel three-dimensional stimuli for a brief baseline period before a researcher demonstrates a series of actions that culminate in an end- or goal-state. The participant is allowed to imitate the demonstrated actions immediately, after a delay, or both. Recall performance is then compared to baseline or to performance on novel control sequences presented at the same session; memory can be assessed for the individual target actions and the order in which they were completed. This procedure is an accepted analogue to the verbal report techniques used with adults, and it has served to establish a solid foundation of the nature of recall memory in infancy and early childhood. In addition, the elicited or deferred imitation procedure has been modified and adapted to answer questions relevant to other aspects of cognitive functioning. The broad utility and application of imitation paradigms is discussed, along with limitations of the approach and directions for future research.

The elicited imitation procedure was established to examine the development of recall memory in infancy and early childhood. This procedure has been widely used to establish a solid foundation of the nature of recall memory in infancy and early childhood.

Undersøgelse Recall Hukommelse i barndom og tidlige barndom Brug af fremkaldte Imitation Paradigm

The ability to recall the past allows us to report on details of previous experiences, from the everyday to the significant. Because recall memory is ...

The Rodent Psychomotor Vigilance Test (rPVT): A Method for Assessing Neurobehavioral Performance in Rats and Mice

The human Psychomotor Vigilance Test (PVT) is a widely used procedure for measuring changes in fatigue and sustained attention. The present article describes a rodent version of the PVT&#8212;termed the &#34;rPVT&#34;&#8212;that measures similar aspects of attention (i.e., performance accuracy, motor speed, premature responding, and lapses in attention). Data are presented that demonstrate both the short- and long-term usefulness of the rPVT when employed with laboratory rats. Rats easily learn the rPVT, and learning to perform the basic procedure takes less than two weeks of training. Once acquired, rat performances in the rPVT show a high degree of similarity to these same performance measures in the human PVT, including similarities in, lapses in attention, reaction times, vigilance decrements across session time (i.e., the human &#34;time-on-task&#34; effects), and the response-stimulus interval (RSI) effect described for humans. Thus the rPVT can be an extremely valuable tool for assessing the effects of a wide range of variables on sustained attention quite similar to human PVT performances, and thus can be useful for developing novel treatments for neurobehavioral dysfunctions.

The Rodent Psychomotor Vigilance Test rPVT: A Method for Assessing Neurobehavioral Performance in Rats and Mice

A rat version of the human Psychomotor Vigilance Test (PVT) is described that measures aspects of attention similar to those measured with the human PVT, including aspects of human vigilance such as performance accuracy, motor speed, premature responding, and lapses in attention.

Den gnaver Psykomotorisk Årvågenhed Test (rPVT): en metode til vurdering neurobehavioral Performance i rotter og mus

The human Psychomotor Vigilance Test (PVT) is a widely used procedure for measuring changes in fatigue and sustained attention. The present article ...

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 ...

A High-performance Liquid Chromatography Measurement of Kynurenine and Kynurenic Acid: Relating Biochemistry to Cognition and Sleep in Rats

The kynurenine pathway (KP) of tryptophan degradation has been implicated in psychiatric disorders. Specifically, the astrocyte-derived metabolite kynurenic acid (KYNA), an antagonist at both N-methyl-d-aspartate (NMDA) and &#945;7 nicotinic acetylcholine (&#945;7nACh) receptors, has been implicated in cognitive processes in health and disease. As KYNA levels are elevated in the brains of patients with schizophrenia, a malfunction at the glutamatergic and cholinergic receptors is believed to be causally related to cognitive dysfunction, a core domain of the psychopathology of the illness. KYNA may play a pathophysiologically significant role in individuals with schizophrenia. It is possible to elevate endogenous KYNA in the rodent brain by treating animals with the direct bioprecursor kynurenine, and preclinical studies in rats have demonstrated that acute elevations in KYNA may impact their learning and memory processes. The current protocol describes this experimental approach in detail and combines a) a biochemical analysis of blood kynurenine levels and brain KYNA formation (using high-performance liquid chromatography), b) behavioral testing to probe the hippocampal-dependent contextual memory (passive avoidance paradigm), and c) an assessment of sleep-wake behavior [telemetric recordings combining electroencephalogram (EEG) and electromyogram (EMG) signals] in rats. Taken together, a relationship between elevated KYNA, sleep, and cognition is studied, and this protocol describes in detail an experimental approach to understanding function outcomes of kynurenine elevation and KYNA formation in vivo in rats. Results obtained through variations of this protocol will test the hypothesis that the KP and KYNA serve pivotal roles in modulating sleep and cognition in health and disease states.

Alterations in the kynurenine pathway (KP) neuroactive metabolites are implicated in psychiatric illnesses. Investigating the functional outcomes of an altered kynurenine pathway metabolism in vivo in rodents may help elucidate novel therapeutic approaches. The current protocol combines biochemical and behavioral approaches to investigate the impact of an acute kynurenine challenge in rats.

En High-performance væskekromatografi måling af Kynurenine og Kynurenic syre: vedrørende biokemi kognition og søvn hos rotter

The kynurenine pathway (KP) of tryptophan degradation has been implicated in psychiatric disorders. Specifically, the astrocyte-derived metabolite ...

A Conditioned Place Preference Protocol for Measuring Incubation of Craving in Rats

A major cause of repeated relapses is a craving for the drug. Drug craving increases progressively during the abstinence period, a phenomenon termed incubation of drug craving. Here, we describe a morphine conditioned place preference (CPP) protocol for measuring the incubation of craving in rats. In this protocol, a CPP paradigm mainly employing somatosensory cues is used to establish a long-term reward memory of morphine. A three-chamber CPP box that differs in the texture of the chamber floor is constructed. First, the animals are tested for their baseline preference to the two side chambers for three consecutive days. Then, they are injected intraperitoneally with morphine/saline and put into their non-preferred/preferred chamber for 45 min. After 6 days of conditioning, their preference to the side chambers is tested for 15 min at different time points after the last conditioning session. With this paradigm, the reward memory of morphine could last for at least 18 days. To test whether the above-mentioned protocol can model increased craving, the number of entrances into the two side chambers are counted during the abstinence period. The results show that the entrances increased, suggesting that the CPP paradigm could mimic the incubation of craving. Future studies can employ this model to study neural mechanisms underlying long-term memory and incubation of craving.

Here, a morphine conditioned place preference (CPP) protocol is described to measure incubation of craving in rats.

En konditioneret sted præference protokol til måling af inkubation af trang hos rotter

A major cause of repeated relapses is a craving for the drug. Drug craving increases progressively during the abstinence period, a phenomenon termed ...