This work was supported by The National Institute of Health (NIH-2RO1NS033661) and by the Alabama Agricultural Experiment Station (HATCH ALA021-1-09017).

HINWEIS: Alle Verfahren hier durchgeführt wurden eingereicht und von IACUC (Office of Research Compliance) zugelassen und wurden nach NIH-Richtlinien durchgeführt. Mäuse in der Verhaltenstests Paradigma waren naiv und nicht für andere Tests verwendet. Die C57BL / 6-Wildtyp und Knock Out Mäusen in diesem Protokoll verwendet wurden bereits 9 beschrieben wurden und die hier vorgestellten Daten sind von diesem Manuskript. 1. Vorbereitung der Testraum und Open Field Apparat <ol><li> Verwenden Sie eine Mehrfacheinheit offenen Feld Labyrinth (OFM) aus vier Kammern Aktivität wurde für diese Analyse (Abbildung 1) verwendet. Jede Kammer gemessen 50 cm (Länge) x 50 cm (Breite) x 38 cm (Höhe) und wurde aus weißen hoher Dichte und nicht-porösen Kunststoff. </li><li> Textur der Böden des Labyrinths für Traktion beim Gehen während Labyrinth Wände waren glatt. Maze Quadranten waren völlig leer für den Zweck dieser Prüfung. Unter Berücksichtigung der Rest dieses Protocol, wird eine einzelne Quadrant der oben beschriebenen Labyrinth verwendet werden, die OFM demonstrieren. </li><li> Wischen Sie die Kammer mit einem 95% Ethanol vor der Verwendung und vor der anschließenden Tests keine Hinweise Duft durch den vorherigen Thema Maus nach links entfernen. </li><li> Lassen Sie das Ethanol vollständig verdampfen vor dem Testen Mäusen. Dies kann 5-10 Minuten zwischen den einzelnen Test-Session statt. </li><li> Für diese Analyse mit dem SMART Video-Tracking-Software von Panlab / Harvard Apparatus zu erfassen und Mausbewegung zu bewerten. HINWEIS: Jede kommerzielle Video Tracking-Kamera und Software verwendet, um die Testperson zu verfolgen und zu bewerten ergibt sich aus der offenen Feld Labyrinth werden. Es ist sehr wichtig, dass der Anwender zu verstehen, wie die Kalibrierung und führen Sie das für jede einzelne Analyse-Software. Unabhängig von der Videokamera und Tracking-Software verwendet wird, werden die besten Ergebnisse erhalten, wenn die Kamera und die Software sind nach Angaben des Herstellers geeicht. </li><li> Führen Sie den Test ina Standard beleuchteten Raum in der Lage ist, die das Labyrinth Gerät und den Computer erforderlich ist, um die Software auszuführen. Hängen Sie die Videokamera über dem Labyrinth entweder durch Befestigung an der Decke oder mit einem beliebigen erhöhten Unterstützungssystem, das das Objektiv der Kamera, um das gesamte Labyrinth Bereich (Abbildung 2) sehen können. </li><li> Wie das menschliche Administrator des Tests ist, lesen Sie genügend Platz im Zimmer haben, um sein von den Testpersonen im Labyrinth völlig unbeobachtbar, um nicht das Verhalten der Mäuse beeinflussen. </li></ol> 2. Vorbereiten der Software, um Aktivität zu messen <ol><li> Öffnen Sie die Video-Tracking-Software. </li><li> Sobald die Software geöffnet ist, bewegen Sie den Cursor auf die Option &quot;Single-Motivverfolgung&quot; auf der Registerkarte &quot;Data Acquisition&quot; und Klick, um diese Option zu öffnen befindet. </li><li> Wählen Sie die Option &quot;Static Hintergrund&quot; am unteren Rand des Bildschirms. </li><li> Nach der Auswahl &quot;Static Hintergrund&quot; ist entschiedenn, ist es notwendig, die Software zu benutzen, um ein Bild des Labyrinths vor der Zugabe der Testpersonen nehmen. Um dies zu tun, bewegen Sie den Cursor auf &quot;Foto&quot; -Button am unteren Rand des Bildschirms und Klick entfernt. HINWEIS: Die Software wird ein Bild von dem Szenario ohne die Testperson, die aus dem bei der Tracking-Prozess genommen Bild subtrahiert werden zu nehmen. Dies führt nur die Bewegung des Subjekts von der Software analysiert. </li><li> Bestätigen Sie, dass das Hintergrundbild oben genommen vollständig von der Tracking-Software entfernt werden, indem Sie den Cursor auf die Schaltfläche &quot;Test&quot; am unteren Rand des Bildschirms befindet, und klicken Sie einmal. Eine solide weißen Feld wird angezeigt, wenn das Hintergrundbild komplett aus dem Tracking-Bild entfernt werden. Wenn Lichtverhältnisse ändern oder das Labyrinth versehentlich bewegt wird, werden Sie schwarze &quot;Schatten&quot; in diesem Bereich zu sehen was die beiden Bilder nicht perfekt überein. Um dieser Situation abzuhelfen, wiederholen Sie einfach steS. 2.4. </li><li> Nach der Bestätigung, Hintergrundeinstellungen, verwenden Sie die Option Zeitplan, um den Weg Zeit wird während der Aufnahme gesteuert konfigurieren. Dazu bewegen Sie den Cursor auf das Register &quot;Konfiguration&quot; und klicken Sie einmal auf die &quot;Timings&quot; Überschrift. Verwenden Sie den neu geöffneten Fenster auf experimentelle Parameter eingeben. </li><li> Aus diesem Protokoll, wählte die &quot;programmierte Zeit&quot; Option als 10 Minuten Tracking Zeitraum. Stellen Sie die &quot;Latenzzeit&quot;, um 5 Sekunden, um dem Benutzer Zeit zu ermöglichen, um die Maus in die Mitte des Irrgartens zu platzieren und weg vor dem Beginn der Verfolgung. Geben Sie die &quot;Aufnahmezeit&quot; von 10 Minuten für die Dauer des Tests. Stellen Sie die &quot;Stop-Steuerung&quot; auf &quot;Beim programmierten Zeit (10 min) ist vorbei&quot;, die automatisch ausgeschaltet, wird die Kamera und die Tracking-Funktion der Software. </li><li> Bewegen Sie den Cursor auf die Schaltfläche &quot;Schließen&quot;, wenn alle Zeiten festgelegt wurden, um das Fenster zu schließen. Sie sind nun bereit, um den Test zu startening Verfahren. </li></ol> 3. Verwaltung der Open Field Test ANMERKUNG: Das Softwarepaket in diesem Protokoll verwendet ermöglicht die Verfolgung von bis zu 16 einzelnen Mäusen zu einem Zeitpunkt. Zur Erleichterung der Fertigstellung und wie oben erwähnt, ist die hier diskutierten Protokoll für eine einzelne Maus mit einem einzigen Quadranten des OFM. Für die verwendeten Geräte für dieses Protokoll, könnte ein Maximum von 4 einzelnen Mäusen mit jedem Quadranten des Labyrinths verfolgt werden. Bei Verwendung einer Multi-Gehäuse Labyrinth, nachdem Sie das erste Thema der Maus in seiner definierten Quadranten, legen Sie die restlichen Mäuse in ihre jeweiligen Labyrinth Quadranten für die Verfolgung Analyse. Für die Zwecke dieses Protokolls wird weitere Anweisung auf einen einzigen Quadranten des Labyrinths ist. <ol><li> Bringen Sie die Mäuse in ihre Käfige aus ihren Käfigen Raum in den Testraum. Den Mäusen an den Eingriffsraum für mindestens 30 Minuten vor Beginn des Tests zu akklimatisieren. </li><li> Entfernen eines einzelnen mOuse aus dem Käfig durch leichtes Greifen mit dem Schwanz und platzieren Sie den Mauszeiger in die Mitte des offenen Feld Labyrinth, während gleichzeitig die Aktivierung der SMART-Software durch einfaches Klicken auf die Schaltfläche Start Tracking Mausbewegung zu beginnen. Es ist normal, dass die Maus, um sofort an die Peripherie Wände des Labyrinths und der Zeitpunkt der Freigabe zu bewegen und Tracking Eroberung der Maus sollten übereinstimmen, um diese Bewegung aufzunehmen. </li><li> Lassen Sie freien und ununterbrochenen Bewegung des Motivs Maus während des jeweiligen Quadranten des Labyrinths für einen einzelnen Zeitraum von 10 Minuten während dieser Zeit wird der Tracking-Software-Bewegung aufzeichnen (Abbildung 3). </li><li> Am Ende des Testzeitraums, nehmen Sie den Betreff Maus sanft, sie aus dem Labyrinth zu entfernen und sie wieder in ihre Heimat Käfig. </li><li> Vor der Reinigung des Labyrinths, visuell zählen die fäkale Boli in das Labyrinth vorhanden Pellets und manuell erfassen die Zahlen für die weitere Analyse. </li><li> Entfernen Sie alle fäkalen Pellets und wischen Sie alle Spots des Wasserlassens. Sprühen Sie den Boden und die Wände des Labyrinths Quadranten mit 95% Ethanol und wischen Sie mit einem sauberen Papiertuch. Lassen Sie die Ethanollösung vollständig trocknen, bevor die Prüfung andere Mäuse. </li><li> Wiederholen Sie den Vorgang mit dem nächsten Maus. </li></ol> 4. Messung und Analyse von Verhalten bei Testverfahren HINWEIS: Bei der Messung werden drei Aspekte der offenen Feldverhalten ohne weiteres mit diesem Protokoll (siehe Diskussion) gekennzeichnet. Eine kurze Anleitung, wie man diese Messungen in der Video-Tracking-Software zugreifen folgt. <ol><li> Wählen Sie im Hauptbildschirm des SMART-Software, bewegen Sie den Cursor auf die &quot;Zones&quot; Menü und Klick &quot;Definition&quot;, um die Zonen-Editor zu öffnen. </li><li> Folgen Sie den detaillierten Anweisungen in der SMART-Software Anwenderhandbuch nicht festlegen Zonen oder Gitter auf den Tracking-Wege zu überlagern. Hier wurde die Software verwendet, um eine 5 x 5 Raster von 10 cm Quadrate für den Boden des das Labyrinth (define <strong&gt; 4). Achten Sie darauf, die Zonendatei erstellt vor dem Schließen der Zonen-Editor speichern. </li><li> Wählen Sie im Hauptbildschirm des SMART-Software, bewegen Sie den Cursor auf die Registerkarte &quot;Analysis&quot; und Einzel klicken, um die Datenanalyse-Fenster zu öffnen. </li><li> Bewegen Sie den Cursor auf die Registerkarte &quot;Datei&quot; und öffnen Sie die Zone-Datei oben erstellt. </li><li> Bewegen Sie den Cursor auf das Register &quot;Konfiguration&quot; und öffnen Sie die Option &quot;Track-Analyse&quot;. Das Fenster &quot;Fach-Analyse-Konfiguration&quot; zu öffnen. <ol><li> Bewegen Sie den Cursor auf die Registerkarte &quot;Standard&quot; und bewegen Sie den Parameter &quot;Wegstrecke&quot; aus der Liste Verfügbare Parameter Box (linke Seite), um die Parameter enthalten Feld (rechte Seite). </li><li> Bewegen Sie den Cursor auf die Registerkarte &quot;Zonen Übergänge&quot; und stellen Sie die erforderlichen Parameter für die Parameter enthalten Box wie oben. </li><li> Stellen Sie sicher, die &quot;Full Track&quot; Box wird am unteren Rand des Fensters überprüft. </li><li> Mohabe den Cursor auf die Schaltfläche OK, und schließen Sie das Fach-Analyse Konfigurationsfenster. HINWEIS: Je nach der Analyse die Sie ausführen möchten, können viele andere Optionen in diesem Fenster zu mir Daten aus der Analyse ausgewählt werden. Lesen Sie die ausführliche Bedienungsanleitung Ihres spezifischen Programms zu ermitteln, welche Parameter für die Analyse Ihrer Daten sehr wichtig. </li></ol></li><li> Unter der Registerkarte Datei in der oberen linken Rand des Programmfensters, öffnen Sie die Einzel Thema Trackfenster und setzen Sie ein Häkchen neben alle Spuren analysiert. Bewegen Sie den Cursor auf die Schaltfläche Check Mark am oberen Rand des Fensters und klicken Sie auf die Track-Explorer-Fenster zu schließen. </li><li> Bewegen Sie den Cursor auf die Schaltfläche &quot;Go&quot; in der Datenanalyse-Fenster und klicken Sie, um Einzelanalyse der Track-Daten zu starten. </li><li> Analysedaten können entweder als ASCII-Textdateien ausgegeben werden, oder es kann direkt in eine Excel-Tabelle exportiert werden. Verwenden Sie die Ausgabe-Tools der Software-Programm Sie verwenden, um die Ausgabe data für den eigenen Gebrauch. HINWEIS: der gesamten Fahrleistung und Zeit im angegebenen Zonen verbracht werden ausgegeben im Anschluss an die Datenanalyse oben beschriebenen Schritte sein. Auch hier wird betont, dass die Schritte, die Messungen hier dargestellt in Abhängigkeit von der Anwendersoftware verwendet werden, um unterscheiden zu erreichen. Aber die Daten selbst und der Interpretation der Ergebnisse sollte ähnlich unabhängig von der Software verwendet wird. Es ist auch erwähnenswert, dass Testleiter Vorspannung von diesem Protokoll als alle gesammelt, quantifiziert Daten von der Software und nicht die des Administrators Messdaten entfernt. Somit besteht keine qualifizierbare Element auf die gesammelten Daten, wie beschrieben. </li></ol>

<ol>
	<li>Hall, C. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Emotional+behavior+in+the+rat:+defecation+and+urination+as+measures+of+individual+differences+in+emotionality.">Emotional behavior in the rat: defecation and urination as measures of individual differences in emotionality.</a> J. Comp. Psychol. 18, 385-403 (1934).</li><li>Walsh, R. N., Cummins, R. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+open+field+test:+a+critical+review.">The open field test: a critical review.</a> Psychol. Bull. 83, 482-504 (1976).</li><li>Prut, L., Belzung, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+open+field+as+a+paradigm+to+measure+the+effects+of+drugs+on+anxiety-like+behaviors:+a+review.">The open field as a paradigm to measure the effects of drugs on anxiety-like behaviors: a review.</a> Eur. J. of Pharm. 463, 3-33 (2003).</li><li>Choleris, E., Thomas, A. W., Kavaliers, M., Prato, F. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+detailed+ethological+analysis+of+the+mouse+open+field+test:+effects+of+diazepam,+chlordiazpoxide+and+an+extremely+low+frequency+pulsed+magnetic+field.">A detailed ethological analysis of the mouse open field test: effects of diazepam, chlordiazpoxide and an extremely low frequency pulsed magnetic field.</a> Neurosci. Biobehav. Rev. 25, 235-260 (2001).</li><li>Ramos, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Animal+Models+of+anxiety:+do+I+need+multiple+tests.">Animal Models of anxiety: do I need multiple tests.</a> TIPS. 29, 493-498 (2008).</li><li>Archer, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Tests+for+emotionality+in+rats+and+mice:+a+review.">Tests for emotionality in rats and mice: a review.</a> Anim. Behav. 21, 205-235 (1973).</li><li>Gray, J. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Emotionality+in+male+and+female+rodents:+a+reply+to+Archer.">Emotionality in male and female rodents: a reply to Archer.</a> Brit. J. Psych. 70, 425-440 (1979).</li><li>Carola, V., D&#8217;Olimpio, F., Brunamonti, E., Mangia, F., Renzi, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Evaluation+of+the+elevated+plus-maze+and+open-field+tests+for+the+assessment+of+anxiety-related+behavior+in+inbred+mice.">Evaluation of the elevated plus-maze and open-field tests for the assessment of anxiety-related behavior in inbred mice.</a> Behav. Brain Res. 134, 49-57 (2002).</li><li>Ramesh Babu, J., Seibenhener, M. L., Peng, J., Strom, A. L., Kemppainen, R., Cox, N., Zhu, H., Wooten, M. C., Diaz-Meco, M. T., Moscat, J., Wooten, M. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Genetic+inactivation+of+p62+leads+to+accumulation+of+hyperphosphorylated+tau+and+neurodegeneration.">Genetic inactivation of p62 leads to accumulation of hyperphosphorylated tau and neurodegeneration.</a> J. Neurochem. 106, 107-120 (2008).</li><li>Simon, P., Dupuis, R., Costentin, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Thigmotaxis+as+an+index+of+anxiety+in+mice:+influence+of+dopaminergic+transmissions.">Thigmotaxis as an index of anxiety in mice: influence of dopaminergic transmissions.</a> Behav. Brain Res. 61, 59-64 (1994).</li><li>Denenberg, V. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Open-field+behavior+in+the+rat:+what+does+it+mean.">Open-field behavior in the rat: what does it mean.</a> Ann. N.Y. Acad. Sci. 159, 852-859 (1969).</li><li>Stanford, S. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+open+field+test:+reinventing+the+wheel.">The open field test: reinventing the wheel.</a> J. Psychopharm. 21, 134-135 (2007).</li><li>Ennaceur, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Tests+of+unconditional+anxiety+&#8211;+pitfalls+and+disappointments.">Tests of unconditional anxiety &#8211; pitfalls and disappointments.</a> J. Phys. Behav. 135, 55-71 (2014).</li><li>Borta, A., Schwarting, R. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Inhibitory+avoidance,+pain+reactivity,+and+plus-maze+behavior+in+Wistar+rats+with+high+versus+low+rearing+activity.">Inhibitory avoidance, pain reactivity, and plus-maze behavior in Wistar rats with high versus low rearing activity.</a> J. Phys. Behav. 84, 387-396 (2005).</li><li>Costall, B., Jones, B. J., Kelly, M. E., Naylor, R. J., Tomkins, D. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Exploration+of+mice+in+a+black+and+white+test+box:+validation+as+a+model+of+anxiety.">Exploration of mice in a black and white test box: validation as a model of anxiety.</a> Pharmacol. Biochem. Behav. 32, 777-785 (1989).</li><li>Bouwknecht, J. A., Paylor, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pitfalls+in+the+interpretation+of+genetic+and+pharmacological+effects+on+anxiety-like+behavior+in+rodents.">Pitfalls in the interpretation of genetic and pharmacological effects on anxiety-like behavior in rodents.</a> Behav. Pharm. 19, 385-402 (2008).</li><li>Webster, D. G., Baumgardner, D. J., Dewsbury, D. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Open+field+behavior+in+eight+taxa+of+muriod+rodents.">Open field behavior in eight taxa of muriod rodents.</a> Bull. Psychonom. Soc. 13, 90-92 (1979).</li><li>Wilson, R. C., Vacek, T., Lanier, D. L., Dewsbury, D. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Open+field+behavior+in+muroid+rodents.">Open field behavior in muroid rodents.</a> Behav. Biol. 17, 495-506 (1976).</li><li>Crawley, J. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Behavioral+phenotyping+of+transgenic+and+knockout+mice:+experimental+designs+and+evaluation+of+general+health,+sensory+functions,+motor+abilities+and+specific+behavioral+tests.">Behavioral phenotyping of transgenic and knockout mice: experimental designs and evaluation of general health, sensory functions, motor abilities and specific behavioral tests.</a> Brain Res. 835, 18-26 (1999).</li><li>Simon, P., Dupuis, R., Costentin, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Thigmotaxis+as+an+index+of+anxiety+in+mice.+influence+of+dopaminergic+transmissions.">Thigmotaxis as an index of anxiety in mice. influence of dopaminergic transmissions.</a> Behav. Brain Res. 61, 59-64 (1994).</li><li>Miller, B. H., Schultz, L. E., Gulati, A., Su, A. I., Pletcher, M. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Phenotypic+characterization+of+a+genetically+diverse+panel+of+mice+for+behavioral+despair+and+anxiety.">Phenotypic characterization of a genetically diverse panel of mice for behavioral despair and anxiety.</a> PLoS One. 5, e14458 (2010).</li><li>Tannenbaum, B., Anisman, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Impact+of+chronic+intermittent+challenges+in+stressor-susceptible+and+resilient+strains+of+mice.">Impact of chronic intermittent challenges in stressor-susceptible and resilient strains of mice.</a> Biol. Psych. 53, 292-303 (2003).</li><li>Lister, R. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ethologically-based+animal+models+of+anxiety+disorders.">Ethologically-based animal models of anxiety disorders.</a> Pharmacol. Ther. 46, 321-340 (1990).</li><li>Kulesskaya, N., Voikar, V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Assessment+of+mouse+anxiety-like+behavior+in+the+light-dark+box+and+open-field+arena:+role+of+equipment+and+procedure.">Assessment of mouse anxiety-like behavior in the light-dark box and open-field arena: role of equipment and procedure.</a> Phys. Behav. 133, 30-38 (2014).</li><li>Han, H., Du, W., Zhou, B., Zhang, W., Xu, G., Niu, R., Sun, Z. <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+chronic+fluoride+exposure+on+object+recognition+memory+and+mRNA+expression+of+SNARE+complex+in+hippocampus+of+male+mice.">Effects of chronic fluoride exposure on object recognition memory and mRNA expression of SNARE complex in hippocampus of male mice.</a> Biol. Trace Elem. Res. 158, 58-64 (2014).</li><li>Barrow, P., Leconte, I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+influence+of+body+weight+on+open+field+and+swimming+maze+performance+during+the+post-weaning+period+in+the+rat.">The influence of body weight on open field and swimming maze performance during the post-weaning period in the rat.</a> Lab. Animals. 30, 22-27 (1996).</li></ol>

The authors have nothing to disclose.

Das Freie Feld Maze ist eine der am weitesten verbreiteten Plattformen in Tierverhaltensstudien. Eine Reihe wichtiger konventionellen und ethologischer Parameter 2,4 gesammelt und während der Durchführung des OFM analysiert werden. Diese Daten ermöglichen dem Forscher Verhalten von Gesamtbewegungsaktivität, Angst bezogenen emotionalen Verhaltensweisen 8 zu messen. Jedoch ist die Verwendung des OFM nicht ohne Nachteile. Eine verwirrende Frage ist die breite Palette von statischen Variablen, die ...

The Open Field Maze (OFM) was initially developed in 1934 as a test to measure emotionality in rodents1. It has attained the status of being one of the most widely used measures of behavior in animal psychology2. It provides an easy and fairly rapid assessment of well-defined behaviors requiring no training to the test subject and little to no specialized training for the human administering the test. These attributes have led to wide-spread use of the open field maze in research extended to other animal species such as calves, pigs, rabbits, primates, honeybees and lobsters3. Part of its popularity arises from the fact that the psychological and physiological concepts underlying the tests are generally straight-forward and well understood. For example, it has been postulated that evolutionary forces have selected for a common response in animals such that most species display anxiety-mediated fear or flight responses to specific stimuli. Rodents for example, show distinct aversions to large, brightly lit, open and unknown environments4. We can assume they have been phylogenetically conditioned to see these types of environments as dangerous. All of these features are incorporated in the open field maze and form the basis of its use in behavioral paradigm testing.
An open field maze consists of a wall-enclosed area that is of sufficient height to prevent the subject from escaping. Typical maze shapes are circular or square with an area large enough, based on the size of the subject tested, to elicit a feeling of openness in the center of the maze. A number of variables can be scored in the open field maze with most parameters involving differing types of motor activity2. Ambulation is the most common behavior studied but others such as latency or rearing can also be measured. Most often, rodent behavior is analyzed in a bare maze. However, the addition of objects, either one or many to the maze floor, adds the ability to see how the subject interacts with novel additional stimuli2. Relevant parameters when objects are presented are typically the number of approaches to an object or in some cases, preference or aversion for one object over another.
Many behavioral tests of anxiety are based on the subject animal&#8217;s body activity and locomotion5. Interpreting behavioral tests for emotionality while separating non-emotional confounding factors, such as motor activity, has been the subject of intense debate6,7. As the OFM was originally described, two measures of emotionality can be deduced, locomotor activity and fecal boli deposits or defecation1. However, these two measures have been shown in some studies to be unrelated supporting the conclusion that emotionality in rodents is multidimensional5. Regardless, discrepancies in the literature regarding these measures and emotionality or anxiety in mouse models may be attributed to differences in analysis criteria or differences in testing procedures. Studies have conclusively linked results from OFM analysis with other measures of anxiety when comparing mouse models8.

<table><tbody><tr><td>Multi Unit Open Field Test</td><td>San Diego Instruments, Inc.</td><td>White 7001-0354</td><td>Any single or multi unit open field maze can be used</td></tr><tr><td>SMART DT Tracking Software</td><td>PanLab/Harvard Apparatus</td><td>76-0695</td><td>Any tracking software can be utilized with this protocol</td></tr><tr><td>Sony 990x Video Camera Recorder</td><td>Sony</td><td>CCD-TRV328</td><td>Any suitable video camera can be attached to computer for recording tracking profiles.</td></tr></tbody></table>

use of the open field maze to measure locomotor and anxiety-like behavior in mice

Animal models have proven to be invaluable to researchers trying to answer questions regarding the mechanisms of behavior. The Open Field Maze is one of the most commonly used platforms to measure behaviors in animal models. It is a fast and relatively easy test that provides a variety of behavioral information ranging from general ambulatory ability to data regarding the emotionality of the subject animal. As it relates to rodent models, the procedure allows the study of different strains of mice or rats both laboratory bred and wild-captured. The technique also readily lends itself to the investigation of different pharmacological compounds for anxiolytic or anxiogenic effects. Here, a protocol for use of the open field maze to describe mouse behaviors is detailed and a simple analysis of general locomotor ability and anxiety-related emotional behaviors between two strains of C57BL/6 mice is performed. Briefly, using the described protocol we show Wild Type mice exhibited significantly less anxiety related behaviors than did age-matched Knock Out mice while both strains exhibited similar ambulatory ability.

Die durchschnittliche Zahl der Individuen pro Stamm von Mäusen in den meisten Fällen etwa 20 getestet, um eine ausreichende statistische Relevanz zu erzeugen. Jedoch kann diese Zahl im Bereich von 8-30 je Maus Verfügbarkeit. In Abhängigkeit von der Messung oder Vergleiche benötigt werden, ist es auch günstig, altersangepassten Probanden verwenden. Die erste und wohl wichtigste spezifische Parameter im Open Field Maze messen Gesamt ambulante Entfernung. Während die Messeinheit irreleva...

Watch this Scientific Journal Video about Use of the Open Field Maze to Measure Locomotor and Anxiety-like Behavior in Mice at JoVE.com

Use of the Open Field Maze to Measure Locomotor and Anxiety-like Behavior in Mice

A protocol is provided to use an Open Field Maze to access general locomotor activity, anxiety and emotionality in a laboratory mouse model.

Die Nutzung der Open Field Maze auf Bewegungsapparat und Angst-ähnliches Verhalten bei Mäusen messen

Animal models have proven to be invaluable to researchers trying to answer questions regarding the mechanisms of behavior. The Open Field Maze is one of ...

use-open-field-maze-to-measure-locomotor-anxiety-like-behavior

Universidad San Sebastian

Research

JoVE Journal

Behavior

72.4K Aufrufe.  Auburn University. A protocol is provided to use an Open Field Maze to access general locomotor activity, anxiety and emotionality in a laboratory mouse model.

Serie: Use of the Open Field Maze to Measure Locomotor and Anxiety-like Behavior in Mice

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.

Die sukzessive Gassen Test von Angst bei Mäusen und Ratten

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.  ...

Forschung

Verhalten

Low-stress Route Learning Using the Lashley III Maze in Mice

Many behavior tests designed to assess learning and memory in rodents, particularly mice, rely on visual cues, food and/or water deprivation, or other aversive stimuli to motivate task acquisition. As animals age, sensory modalities deteriorate. For example, many strains of mice develop hearing deficits or cataracts. Changes in the sensory systems required to guide mice during task acquisition present potential confounds in interpreting learning changes in aging animals. Moreover, the use of aversive stimuli to motivate animals to learn tasks is potentially confounding when comparing mice with differential sensitivities to stress. To minimize these types of confounding effects, we have implemented a modified version of the Lashley III maze. This maze relies on route learning, whereby mice learn to navigate a maze via repeated exposure under low stress conditions, e.g. dark phase, no food/water deprivation, until they navigate a path from the start location to a pseudo-home cage with 0 or 1 error(s) on two consecutive trials. We classify this as a low-stress behavior test because it does not rely on aversive stimuli to encourage exploration of the maze and learning of the task. The apparatus consists of a modular start box, a 4-arm maze body, and a goal box. At the end of the goal box is a pseudo-home cage that contains bedding similar to that found in the animal&#x2019;s home cage and is specific to each animal for the duration of maze testing. It has been demonstrated previously that this pseudo-home cage provides sufficient reward to motivate mice to learn to navigate the maze1. Here, we present the visualization of the Lashley III maze procedure in the context of evaluating age-related differences in learning and memory in mice along with a comparison of learning behavior in two different background strains of mice. We hope that other investigators interested in evaluating the effects of aging or stress vulnerability in mice will consider this maze an attractive alternative to behavioral tests that involve more stressful learning tasks and/or visual cues.

The Lashley III maze is a route-learning task that does not rely on aversive stimuli or visual cues. It is thus a highly attractive option for evaluating learning and memory, especially in aging mice or otherwise where stress is a consideration.

Low-Stress-Route Learning Mit dem Lashley III Maze in Mäuse

Many behavior tests designed to assess learning and memory in rodents, particularly mice, rely on visual cues, food and/or water deprivation, or other ...

Neurowissenschaften

Barnes Maze Testing Strategies with Small and Large Rodent Models

Spatial learning and memory of laboratory rodents is often assessed via navigational ability in mazes, most popular of which are the water and dry-land (Barnes) mazes. Improved performance over sessions or trials is thought to reflect learning and memory of the escape cage/platform location. Considered less stressful than water mazes, the Barnes maze is a relatively simple design of a circular platform top with several holes equally spaced around the perimeter edge. All but one of the holes are false-bottomed or blind-ending, while one leads to an escape cage. Mildly aversive stimuli (e.g.&#160;bright overhead lights) provide motivation to locate the escape cage. Latency to locate the escape cage can be measured during the session; however, additional endpoints typically require video recording. From those video recordings, use of automated tracking software can generate a variety of endpoints that are similar to those produced in water mazes (e.g.&#160;distance traveled, velocity/speed, time spent in the correct quadrant, time spent moving/resting, and confirmation of latency). Type of search strategy (i.e.&#160;random, serial, or direct) can be categorized as well. Barnes maze construction and testing methodologies can differ for small rodents, such as mice, and large rodents, such as rats. For example, while extra-maze cues are effective for rats, smaller wild rodents may require intra-maze cues with a visual barrier around the maze. Appropriate stimuli must be identified which motivate the rodent to locate the escape cage. Both Barnes and water mazes can be time consuming as 4-7 test trials are typically required to detect improved learning and memory performance (e.g.&#160;shorter latencies or path lengths to locate the escape platform or cage) and/or differences between experimental groups. Even so, the Barnes maze is a widely employed behavioral assessment measuring spatial navigational abilities and their potential disruption by genetic, neurobehavioral manipulations, or drug/ toxicant exposure.

The dry-land Barnes maze is widely used to measure spatial navigation ability in response to mildly aversive stimuli. Over consecutive days, performance (e.g.&#160;latency to locate escape cage) of control subjects improves, indicative of normal learning and memory. Differences between rats and mice necessitate apparatus and methodology changes that are detailed here.

Barnes Maze Teststrategien mit kleinen und großen Nager-Modelle

Spatial learning and memory of laboratory rodents is often assessed via navigational ability in mazes, most popular of which are the water and dry-land ...

Place and Response Learning in the Open-field Tower Maze

This protocol describes how the Open-field Tower Maze (OFTM) paradigm is used to study spatial learning in rodents. This maze is especially useful for examining how rats learn to use a place- or response-learning to successfully navigate in an open-field arena. Additionally, this protocol describes how the OFTM differs from other behavioral maze paradigms that are commonly used to study spatial learning in rodents. The OFTM described in this article was adapted from the one previously described by Cole, Clipperton, and Walt (2007). Specifically, the OFTM was created to test spatial learning in rodents without the experimenter having to consider how &#8220;stress&#8221; might play a role as a confounding variable. Experiments have shown that stress-alone can significantly affect cognitive function1. The representative results section contains data from an experiment that used the OFTM to examine the effects of estradiol treatment on place- and response-learning in adult female Sprague Dawley rats2. Future studies will be designed to examine the role of the hippocampus and striatum in place- and response-learning in the OFTM.

The Open-field Tower Maze (OFTM) is a maze created to study the behavioral and neural mechanisms of spatial learning (e.g., place- or response-learning) in rats. This maze is especially useful for experimenters who want to use a non-stressful maze paradigm to investigate spatial learning in their research.

Zeigen and Response Learning in der Open-field-Turm Maze

This protocol describes how the Open-field Tower Maze (OFTM) paradigm is used to study spatial learning in rodents. This maze is especially useful for ...

Novel Object Exploration as a Potential Assay for Higher Order Repetitive Behaviors in Mice

Restricted, repetitive behaviors (RRBs) are a core feature of autism spectrum disorder (ASD) and disrupt the lives of affected individuals. RRBs are commonly split into lower-order and higher-order components, with lower order RRBs consisting of motor stereotypies and higher order RRBs consisting of perseverative and sequencing behaviors. Higher order RRBs are challenging to model in mice. Current assays for RRBs in mice focus primarily on the lower order components, making basic biomedical research into potential treatments or interventions for higher-order RRBs difficult. Here we describe a new assay, novel object exploration. This assay uses a basic open-field arena with four novel objects placed around the perimeter. The test mouse is allowed to freely explore the arena and the order in which the mouse investigates the novel objects is recorded. From these data, patterned sequences of exploration can be identified, as can the most preferred object for each mouse. The representative data shared here and past results using the novel object exploration assay illustrate that inbred mouse strains do demonstrate different behavior in this assay and that strains with elevated lower order RRBs also show elevated patterned behavior. As such, the novel object exploration assay appears to possess good face validity for higher order RRBs in humans and may be a valuable assay for future studies investigating novel therapeutics for ASD.

Higher order restricted, repetitive behaviors (RRBs) disrupt the lives of affected individuals. These behaviors are challenging to model in rodents, making basic biomedical research into potential treatments or interventions for RRBs difficult. Here we describe novel object exploration as a potential assay for higher order RRBs in mice.

Novel Object Exploration als Potential Assay für höherer Ordnung Repetitive Verhalten bei Mäusen

Restricted, repetitive behaviors (RRBs) are a core feature of autism spectrum disorder (ASD) and disrupt the lives of affected individuals.  RRBs are ...

Elevated Plus Maze for Mice

Although the mouse genome is now completely sequenced, the functions of most of the genes expressed in the brain are not known. The influence of a given gene on a specific behavior can be determined by behavioral analysis of mutant mice. If a target gene is expressed in the brain, behavioral phenotype of the mutant mice could elucidate the genetic mechanism of normal behaviors. The elevated plus maze test is one of the most widely used tests for measuring anxiety-like behavior. The test is based on the natural aversion of mice for open and elevated areas, as well as on their natural spontaneous exploratory behavior in novel environments. The apparatus consists of open arms and closed arms, crossed in the middle perpendicularly to each other, and a center area. Mice are given access to all of the arms and are allowed to move freely between them. The number of entries into the open arms and the time spent in the open arms are used as indices of open space-induced anxiety in mice. Unfortunately, the procedural differences that exist between laboratories make it difficult to duplicate and compare results among laboratories. Here, we present a detailed movie demonstrating our protocol for the elevated plus maze test. In our laboratory, we have assessed more than 90 strains of mutant mice using the protocol shown in the movie. These data will be disclosed as a part of a public database that we are now constructing.
Visualization of the protocol will promote better understanding of the details of the entire experimental procedure, allowing for standardization of the protocols used in different laboratories and comparisons of the behavioral phenotypes of various strains of mutant mice assessed using this test. 


The elevated plus maze test is one of the most widely used tests for measuring anxiety-like behavior in mice. Here, we present a movie showing the detailed procedures for conducting the test.

Elevated Plus Maze für Mäuse

Although the mouse genome is now completely sequenced, the functions of most of the genes expressed in the brain are not known. The influence of a given ...

Biologie

Analyzing Spatial Learning and Prosocial Behavior in Mice Using the Barnes Maze and Damsel-in-Distress Paradigms

The Barnes maze is a reliable measure of spatial learning and memory that does not require food restriction or exposure to extremely stressful stimuli. The Barnes maze can also assess other mouse behaviors, such as general motivation to escape from the maze platform and exploratory behavior. The Barnes maze can measure whether a genetic mutation or environmental variable can impact the acquisition and retention of spatial memories, as well as provide information about the search strategy employed by the mice. Here we use the Barnes maze to detect a memory deficit in adult mice following a single developmental ethanol exposure event. The newly described Damsel-in-Distress paradigm exposes a male mouse to a female mouse trapped in a chamber in the open center field of the arena. It provides an opportunity for the mouse to socially respond to the trapped female and exhibit prosocial behavior. The Damsel-in-Distress paradigm can also be used to examine mouse behavior in a novel arena and measure locomotor activity. Both the Barnes Maze and the Damsel-in-Distress protocols require minimal financial investment and most aspects of the tests can be constructed from common lab supplies. These flexible and accessible tools can also be used to detect behavioral changes over the course of development.

This protocol measures spatial learning and memory using the Barnes maze. A novel Damsel-in-Distress paradigm is used to assess locomotor activity and prosocial behavior in mice.

Analyse der räumlichen Lernens und prosoziales Verhalten bei Mäusen mit dem Barnes Labyrinth und Damsel in Distress Paradigmen

The Barnes maze is a reliable measure of spatial learning and memory that does not require food restriction or exposure to extremely stressful stimuli. ...

A Behavioral Test Battery for the Repeated Assessment of Motor Skills, Mood, and Cognition in Mice

Pharmacological and toxicological studies in neurodegeneration require comprehensive behavioral analysis in mice because motor dysfunctions and dysfunctions in mood and cognition are common and often shared symptoms in neurodegenerative diseases. Shown here is a behavioral test battery for motor, mood, and cognition, which can be repeatedly tested in a longitudinal study. This battery assesses the overall behavioral phenotype in mice by examining each domain of behavior with at least two independent well-accepted tests (i.e., open-field test and rotarod test for motor function, social interaction test, elevated plus maze test, and forced swim test for emotional function, and Morris water maze test and novel object recognition test for cognitive function). Therefore, this sensitive and comprehensive test battery is a powerful tool for the study of behavioral alternation in neurodegeneration.

A comprehensive behavioral test battery of motor skills, mood&#8212;including social interaction, depression, and anxiety&#8212;and cognition is designed for the repeated assessment of neurodegeneration-related behavioral changes in mice.

Eine Verhaltenstherapie Testbatterie für die wiederholte Beurteilung der Motorik, Stimmung und Kognition bei Mäusen

Pharmacological and toxicological studies in neurodegeneration require comprehensive behavioral analysis in mice because motor dysfunctions and ...

Investigating Migraine-Like Behavior Using Light Aversion in Mice

Migraine is a complex neurological disorder characterized by headache and sensory abnormalities, such as hypersensitivity to light, observed as photophobia. Whilst it is impossible to confirm that a mouse is experiencing migraine, light aversion can be used as a behavioral surrogate for the migraine symptom of photophobia. To test for light aversion, we utilize the light/dark assay to measure the time mice freely choose to spend in either a light or dark environment. The assay has been refined by introducing two critical modifications: pre-exposures to the chamber prior to running the test procedure and adjustable chamber lighting, permitting the use of a range of light intensities from 55 lux to 27,000 lux. Because the choice to spend more time in the dark is also indicative of anxiety, we also utilize a light-independent anxiety test, the open field assay, to distinguish anxiety from light-aversive behavior. Here, we describe a modified test paradigm for the light/dark and open field assays. The application of these assays is described for intraperitoneal injection of calcitonin gene-related peptide (CGRP) in two mouse strains and for optogenetic brain stimulation studies.

Rodents are not able to report migraine symptoms. Here, we describe a manageable test paradigm (light/dark and open field assays) to measure light aversion, one of the most common and bothersome symptoms in patients with migraines.

Untersuchung von migräneähnlichem Verhalten mit Lichtaversion bei Mäusen

Migraine is a complex neurological disorder characterized by headache and sensory abnormalities, such as hypersensitivity to light, observed as ...

Validierung eines Mausmodells des Angelman-Syndroms mit Hilfe von Verhaltenstests

Behavioral Characterization of an Angelman Syndrome Mouse Model

This manuscript describes a battery of behavioral tests available to characterize Angelman syndrome (AS)-like phenotypes in an established murine model of AS. We use the rotarod learning paradigm, detailed gait analysis, and nest building test to detect and characterize animal motor impairments. We test animal emotionality in the open field and elevated plus maze tests, as well as the affect in the tail suspension test. When AS mice are tested in the open field test, the results should be interpreted with care, since motor dysfunctions influence mouse behavior in the maze and alter activity scores.
The reproducibility and effectiveness of the presented behavioral tests has already been validated in several independent Uba3a mouse lines with different knockout variants, establishing this set of tests as an excellent validation tool in AS research. Models with the relevant construct and face validity will warrant further investigations to elucidate the pathophysiology of the disease and grant the development of causal treatments.

Autor im Rampenlicht: Eine Reihe von hochgradig reproduzierbaren Verhaltenstests zur Validierung eines Mausmodells mit Angelman-Syndrom 

This manuscript presents a set of highly reproducible behavioral tests to validate an Angelman syndrome mouse model.

Verhaltenscharakterisierung eines Angelman-Syndrom-Mausmodells

This manuscript describes a battery of behavioral tests available to characterize Angelman syndrome (AS)-like phenotypes in an established murine model of ...