Die Autoren danken Herrn Akihiro Hirano für seine Hilfe bei der Herstellung der kundenspezifischen Apparat. Die Autoren möchte Herr Kosuke Takahashi und Herr Shingo Nakajima für ihre technische Unterstützung zu danken. Diese Arbeit wurde durch Forschung an Risiko von chemischen Substanzen, Gesundheit und Labor Sciences Research Grants, Ministerium für Gesundheit, Arbeit und Wohlfahrt, Japan (H25-KAGAKU-IPPANN-003) und Research Project Grant A durch Institut für Wissenschaft und Technologie Meiji unterstützt. Universität, Japan.

Alle hier beschriebene experimentelle Verfahren wurden von den institutionellen Animal Care und Nutzung Ausschuss der Meiji-Universität (IACUC 14-0002(1)). genehmigt Tiere, die in die repräsentativen Ergebnisse waren wie folgt hergestellt: männliche Sprague-Dawley Ratten wurden von einer Labor-Tier-Firma gekauft und befindet sich in Gruppen von 4 Tieren pro standard Rattenkäfig (W200 D410 × × H250 mm) für mindestens eine Woche vor Beginn der der Behavioral Experiments. Die Unterbringung von Tieren-Zimmer war gepflegt, bei standard Umgebungsbedingungen für Licht (12:12 hell/dunkel-Zeitplan mit auf bei 10:00 h leuchtet), Raumtemperatur (25,0 ± 0,5 ° C), Luftfeuchtigkeit (55 ± 10 %) und Lüftung (10 x / h), und Tiere hatte ad libitum Zugang zu Lebensmitteln ( Oriental Yeast Co. Ltd, Tokyo, Japan) und Wasser. Die Tiere waren in einem standard Rattenkäfig (W410 x D250 x H200 mm, Toyoriko Co., Ltd., Aichi, Japan) mit Papier, Bettwäsche (Alpha-Dri, LSG Corporatin, Tokio, Japan) gehalten.
(1) erhöhte Plus Maze Test
<ol><li>Richten Sie die erhöhten Plus Labyrinth Apparat in einem maßgeschneiderten Container (W1, 500 × D1, 500 × H2, 000 mm) (Abbildung 1A) mit einem Plus-förmigen Metallgerüst 500 mm über dem Boden, den Lauf zu unterstützen so Arme (Abbildung 1 b), in der Container-Decke, Beleuchtung beleuchten Sie die Oberfläche des Labyrinths die erforderliche Lichtstärke (Abbildung 1) und eine digitale Videokamera in der Mitte Decke ermöglichen live video-Überwachung der behavioral Tests (Abbildung 1).</li>
	<li>Die Arme (L500 x W100 mm) auf die Plus-förmigen Rahmen befestigen, und dann fix die quadratischen Mittelbahnsteig (W100 x D100 mm) in der Mitte der vier Arme (Abbildung 1 -F). Hinweis: In unserem Labor haben wir zwei Arten von offenen Armen (L500 x W100 mm) und zwei Arten von geschlossenen Arme (L500 x W100 x H450 mm) (Abbildung 1). Die geschlossene Arme haben entweder transparent oder opak Wände (Abbildung 1), und die offenen Arme sollen mit oder ohne einen kurzen Felsvorsprung (H5 mm, Abbildung 1E). Die quadratischen Mittelbahnsteig (W100 × D100 mm) befindet sich im Zentrum der vier Arme (Abbildung 1 -F). So, die folgenden vier Kombinationen von Waffen zubereitet werden, nach experimentellen Zweck: offene Arme mit leisten + geschlossene Arme mit transparenten Wänden (leisten/Transparent), offenen Armen mit leisten + geschlossene Arme mit undurchsichtigen Wänden (leisten/opak), offenen Armen ohne Vorsprünge + geschlossene Arme mit transparenten Wänden (No-leisten/Transparent) und offenen Armen ohne Vorsprünge + geschlossene Arme mit opaker Wände (No-leisten/opak). Die Oberfläche der Start-und Landebahn Arm ist schwarz, um bessere Tier Verfolgung durch die automatisierte Verhalten-Analyse-Software, wie in Abschnitt 2 beschrieben Verhaltensanalyse zu ermöglichen.</li>
	<li>Stellen Sie auf der Oberfläche des distalen Teils jeder Arm und zentrale Plattform bis ca. 100 Lux Beleuchtungsstärke.</li>
	<li>Vor jedem Test den gesamten Apparat mit 70 % Ethanol-Lösung reinigen und mit destilliertem Wasser abwischen.</li>
	<li>Klicken Sie auf "Record-Taste" auf der Harddisk-Recorder, die digitale Videokamera zu starten, die die Ratte Verhalten für das imaging-Analyse beschrieben in Abschnitt 2, Verhaltensanalyse Video aufzeichnen wird.</li>
	<li>Ein Tier auf der zentralen Plattform für einen offenen Arm und freie Erkundung auf dem Gerät für 5 Minuten.</li>
	<li>Klicken Sie auf den Button"Stop" auf der Harddisk-Recorder für die Aufnahme zu beenden.</li>
	<li>Die Anzahl der Kot und Urinieren nach der Rückkehr der getesteten Tieres in der Tierhaltung-Raum.</li>
	<li>Wiederholen Sie ab Schritt 1.4 bis zum Ende des Experiments. Hinweis: Ein trocken Testlauf erfolgt stets vor Beginn jedes Experiment, um zu überprüfen, dass die experimentelle Einstellungen korrekt eingerichtet sind, bevor alle Versuchstiere den experimentellen Bedingungen ausgesetzt sind. Andernfalls kann einige Tiere, besonders das erste Tier unterschiedliche Umweltbedingungen (z.B. Geräusche möglicherweise induziert durch den Experimentator während der Manipulation und/oder Tier Hinweise, die möglicherweise durch das zuvor getestete Tier links) auftreten. Hinweis: Die Experimentatoren sind in der Nähe der Container, wenn ein Tier getestet wird. Obwohl die Experimentatoren nicht sichtbar sind kann die Ratte noch in der Lage, ihnen über Geruch und/oder Ton zu spüren also.</li>
</ol>(2) Verhaltensanalyse
Hinweis: Verhaltensanalyse erfolgt durch manuelle Beobachtung und Analyse-Software automatisiert.
<ol><li>
Manuelle Beobachtung
	<ol>
<li>Nachdem das Tier auf dem Sims platziert haben, beachten Sie Folgendes: Anzahl der Einträge (ein Eintrag auf den Arm ist definiert als jede Instanz, wenn alle 4 Pfoten auf dem Arm sind), verbrachte Zeit an jedem Arm und Anzahl der gestreckt behandelnden Haltungen gegenüber einen offenen Arm , und sichern Sie die zentrale Plattform in Richtung einer geschlossenen Arm.</li>
		<li>Dauer und Häufigkeit der Stoppuhr und Graf für 5 min, ab wann die Türen des Behälters geschlossen sind oben festgelegten Verhaltensweisen zu messen.</li>
	</ol>
</li>
	<li>
Automatisierte Analyse
	<ol>
<li>Mit dem verfügbaren Software-Programm, Folgendes aufzuzeichnen: Abstand reiste, Zeit verbrachte in verschiedene Positionen auf offenen Armen (mittleren proximalen und distalen Segmente von der zentralen Plattform), und eine Analyse von Minute zu Minute der Zeit aufgewendet offen und geschlossen Arme.</li>
		<li>Klicken Sie auf den startButton"" der automatisierten Analyse-Software zur gleichen Zeit, als die manuelle Beobachtung beginnt (die Aufzeichnung stoppt automatisch wenn Aufnahme Dauer festgelegt ist). Hinweis: In dieser Studie verwendete Verhaltensanalyse-Software verfolgt das Tier über den Kontrast mit dem Hintergrund-Feld; das Tier, das eine feste Größe zugeordnet ist, wird als das hellere Objekt identifiziert und die Fortbewegung der Mittelpunkt kann somit für Tier-Tracking verwendet werden.</li>
	</ol>
</li>
</ol>

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L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Behavioral+and+pharmacological+validation+of+the+elevated+plus+maze+constructed+with+transparent+walls.">Behavioral and pharmacological validation of the elevated plus maze constructed with transparent walls.</a> Braz J Med Biol Res. 28 (5), 597-601 (1995).</li><li>Hagenbuch, N., Feldon, J., Yee, B. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Use+of+the+elevated+plus-maze+test+with+opaque+or+transparent+walls+in+the+detection+of+mouse+strain+differences+and+the+anxiolytic+effects+of+diazepam.">Use of the elevated plus-maze test with opaque or transparent walls in the detection of mouse strain differences and the anxiolytic effects of diazepam.</a> Behav Pharmacol. 17 (1), 31-41 (2006).</li><li>Fernandes, C., File, S. E. <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+open+arm+ledges+and+maze+experience+in+the+elevated+plus-maze.">The influence of open arm ledges and maze experience in the elevated plus-maze.</a> Pharmacol Biochem Behav. 54 (1), 31-40 (1996).</li><li>Horii, Y., Kawaguchi, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Higher+detection+sensitivity+of+anxiolytic+effects+of+diazepam+by+ledge-free+open+arm+with+opaque+walled+closed+arm+elevated+plus+maze+in+male+rats.">Higher detection sensitivity of anxiolytic effects of diazepam by ledge-free open arm with opaque walled closed arm elevated plus maze in male rats.</a> Behav Brain Res. 294, 131-140 (2015).</li><li>Santos, R. O., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Evaluation+of+the+effect+of+acute+sibutramine+in+female+rats+in+the+elevated+T-maze+and+elevated+plus-maze+tests.">Evaluation of the effect of acute sibutramine in female rats in the elevated T-maze and elevated plus-maze tests.</a> Basic Clin Pharmacol Toxicol. 114 (2), 181-187 (2014).</li><li>Lynn, D. A., Brown, G. 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Die Autoren haben nichts preisgeben.

Mit steigender Anzahl von Patienten mit psychiatrischen Erkrankungen, einschließlich der Angststörung, ist es wichtig, innovative Studien zur Verbesserung des Verständnisses der Grundmechanismus der Angst entwickeln. Um dies zu erreichen, ist es auch wichtig, die experimentellen Verfahren zu optimieren. Damit zur Verbesserung der Anwendung der erhöhten plus Labyrinth Test, wir haben bewiesen, dass die Wirkung von Arm-Funktionen auf die experimentellen Ergebnisse mit Ratten.
Die erhöhte pl...

Die Zahl von Patienten mit affektiven Störungen, einschließlich-Angststörung und Depression, steigt. Obwohl die Entwicklung dieser Erkrankungen genetische und umweltbedingte Faktoren beinhaltet, sind detaillierte pathogenen Mechanismen noch nicht vollständig verstandenen1. Forschung muss daher konzentrieren sich auf die Aufdeckung der zugrunde liegenden Mechanismen von affektiven Störungen sowie die Entwicklung von wirksamen Medikamenten, die weniger Nebenwirkungen haben.
Verhaltens-Tests für Versuchstiere eignen sich für die Untersuchung von höheren Gehirnfunktion, einschließlich Emotion. Wie das Verhalten der Tiere sehr variabel ist, unterscheiden sich experimentelle Ergebnisse oft zwischen Labors, auch wenn eine identische Inzucht Belastung verwendet2war. Forscher müssen also auch die grundlegenden Funktionen der experimentelle Geräte zu verstehen, die sie ergreifen, um zuverlässige und präzise Ergebnisse im Labor zur Verfügung zu stellen.
Die erhöhte plus Labyrinth Test wurde ursprünglich gegründet, um emotionale Reaktivität in Nagetieren3,4zu prüfen. Den gesamten Apparat in der Regel befindet sich 500 mm über dem Boden und besteht aus zwei offenen und geschlossenen zweiarmig, die eine plus Form zu machen. Messungen der Arm Exploration (einschließlich der Dauer und Häufigkeit der Einträge an jedem Arm), Abstand reiste und bestimmte Verhaltensweisen wie gestreckt behandelnden Haltung des unterworfenen Tiere werden Beobachtungsdaten Zeiten aufgezeichnet. Durch diese Maßnahmen wird die tierische Angst als ein experimentelles Ergebnis geschätzt.
Die erhöhte plus Labyrinth-Test dient in der ganzen Welt, aber nicht alle Labore verwenden die gleichen Apparat Design5,6, und diese Unterschiede können die experimentellen Ergebnisse beeinflussen. Unterschiede sind z. B. das Vorhandensein oder Fehlen von Felsvorsprüngen entlang der offenen Armen und/oder die Nutzung von transparent oder opak Wände für geschlossene Arme5,6. Die Vorsprünge entlang offenen Armen und die Verwendung von transparenten Wänden für geschlossene Arme wurden entwickelt, um Tiere fallen aus dem Apparat7zu verhindern und Verhaltensbeobachtungen innerhalb der geschlossenen Arme, jeweils8,9zu erleichtern. Zwar gab es einige Studien, die wie einzelne Arm Struktur Unterschiede untersucht die tierische Angst1,2,5,10,11 erkennen beeinträchtigen können , die kombinierten Auswirkungen offen (mit/ohne kurze Kante) und geschlossenen (transparenten/undurchsichtigen Wänden) Arm Features auf das Verhalten der Tiere ist noch nicht untersucht.
In einem kürzlich veröffentlichten Bericht diskutierten wir die Wirkung dieser Kombinationen von Arm-Designs auf Angst-ähnliches Verhalten in Ratten12. In Kürze öffnen Sie Arm leisten deutlich erhöhte Angst-ähnliches Verhalten bei Ratten, während transparente Arme können auch dieses Verhalten gesunken. Darüber hinaus überprüft wir höhere Empfindlichkeit des Effekts eine angstlösende Medikament in einer Kombination aus keine-zulasten offenen Arm + opak ummauerten geschlossenen Arm im Vergleich mit einer Kombination aus zulasten offenen Arm + transparente ummauerten geschlossenen Arm. In diesem Bericht werden wir vorstellen unser Protokoll für die erhöhte plus Labyrinth Test, zusammen mit Diskussion der wichtigsten Ergebnisse aus dem vorherigen Bericht. Wir glauben, dass dieser Bericht enthalten nützliche Informationen für diejenigen, die beschäftigt haben oder planen, verwenden Sie die erhöhte plus Labyrinth in ihren Studien.

<table><tbody><tr><td>Customized container (W1500 &amp;times; D1500 &amp;times; H2000 mm)</td><td>NA</td><td>NA</td><td>Handmade by Mr. Akihiro Hirano</td></tr><tr><td>Lighting</td><td>NA</td><td>NA</td><td>Handmade by Mr. Akihiro Hirano</td></tr><tr><td>Elevated plus maze apparatus</td><td>NA</td><td>NA</td><td>Handmade by Mr. Akihiro Hirano</td></tr><tr><td>Digital video camera</td><td>Watec</td><td>WAT-250D2</td><td>Any suitable video camera can be attached to a hard disk recorder for recording tracking profiles.</td></tr><tr><td>Hard disk recorder</td><td>TOSHIBA</td><td>RD-R200</td><td>Any suitable hard disk recorder can be attached to the digital video camera.</td></tr><tr><td>Sprague-Dawley rats</td><td>Charles River Laboratories Japan, Inc.</td><td>NA</td><td>Any suitable rats can be used for your study</td></tr><tr><td>Standard rat cage</td><td>Toyoriko</td><td>TP-105</td><td>Any suitable rat cage be used for your study</td></tr><tr><td>Instrument to measure light intensity</td><td>AS ONE</td><td>1-5818-02</td><td>Any suitable instrument to measure light intensity</td></tr><tr><td>EthoVision XT version 8.0</td><td>Noldus</td><td>NA</td><td>Any suitable software to measure animal behavior</td></tr><tr><td>Diazepam</td><td>Taiyo Pharmaceutical Industry</td><td>1124402A2061</td><td>Any suitable drug can be used for your study</td></tr><tr><td>Tween 80</td><td>Wako Pure Chemical Industries</td><td>9005-65-6</td><td>Any suitable drug can be used for your study</td></tr></tbody></table>

testing animal anxiety in rats: effects of open arm ledges and closed arm wall transparency in elevated plus maze test

Die erhöhte plus Labyrinth-Test ist ein behavioral Test zur Beurteilung tierische Angst bei Nagetieren. Obwohl dieser Test auf dem Gebiet der Verhaltensforschung allgemein angewendet wird, werden oft widersprüchliche Ergebnisse aus verschiedenen Labors angeboten. Ermittlung der Gründe für die unterschiedlichen Ergebnisse, konzentrierten wir uns zuvor auf Arm-Features, die Laboratorien, die meisten vor allem die Anwesenheit/Abwesenheit leisten an den Seiten mit ausgebreiteten Armen und der Transparenz/Undurchsichtigkeit der geschlossenen Arm Wände unterscheiden. In einem früheren Bericht wir verwendet einen benutzerdefinierten gestaltete Container Ratte Verhalten auf verschiedene Kombinationen von offenen und geschlossenen Arm Designs unter sonst gleichen Versuchsbedingungen zu vergleichen, und zeigte, dass Unterschiede im Arm Funktionen experimentelle stören Ergebnisse. In Kürze öffnen Sie Arm leisten deutlich erhöhte Angst-ähnliches Verhalten bei Ratten, während transparente Arme können auch dieses Verhalten gesunken. Darüber hinaus haben wir eine höhere Empfindlichkeit des Effekts eine angstlösende Medikament in einer Kombination aus keine-zulasten offenen Arm + opak ummauerten geschlossenen Arm im Vergleich mit einer Kombination aus zulasten offenen Arm + transparente ummauerten geschlossenen Arm überprüft. In diesem Bericht stellen wir Ihnen unser Protokoll für die erhöhte plus Labyrinth Test, zusammen mit der Diskussion der wichtigsten Ergebnisse aus den vorherigen Bericht und unsere experimentelle Erfahrung. Wir glauben, dass dieser Bericht enthalten nützliche Informationen für Forscher, die beschäftigt haben oder planen, verwenden Sie die erhöhte plus Labyrinth in ihren Studien.

In der vorangegangenen Studie verglichen wir die kombinierte Wirkung der offenen Arm Simsen und Transparenz der geschlossenen Arm Wände auf der explorativen Verhaltensweisen auf der Labyrinth-12. Ratten deutlich viel weniger Zeitaufwand auf offenen Armen ohne Vorsprünge als auf offenen Armen mit leisten (F [1, 60] = 13,49, P = 0,0005) (Abbildung 2A). Ähnliche Ergebnisse wurden auch für die Anzahl der Einträge in offenen A...

Watch this Scientific Journal Video about Testing Animal Anxiety in Rats: Effects of Open Arm Ledges and Closed Arm Wall Transparency in Elevated Plus Maze Test at JoVE.com

Testing Animal Anxiety in Rats: Effects of Open Arm Ledges and Closed Arm Wall Transparency in Elevated Plus Maze Test

Testen tierische Angst bei Ratten: Auswirkungen der offenen Arm Gesimsen und geschlossenen Arm Wand Transparenz in erhöhter Plus Maze Test

Ratten Angst-ähnliches Verhalten getestet durch die erhöhte plus Labyrinth hängt stark von der Testumgebung. Hier zeigen wir, wie Geräte-Einstellungen, besonders Arm Struktur, Verhalten auf das Labyrinth beeinflussen, und wir erklären ein zuverlässiges Protokoll, um Wirkungen angstlösende Medikamente zu erkennen. Dieser Bericht wird Angst Forschung beitragen.

The elevated plus maze test is a behavioral test for assessing animal anxiety in rodents. Although this test is widely applied in the field of behavioral ...

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13.5K Aufrufe.  Meiji University. Ratten Angst-ähnliches Verhalten getestet durch die erhöhte plus Labyrinth hängt stark von der Testumgebung. Hier zeigen wir, wie Geräte-Einstellungen, besonders Arm Struktur, Verhalten auf das Labyrinth beeinflussen, und wir erklären ein zuverlässiges Protokoll, um Wirkungen angstlösende Medikamente zu erkennen. Dieser Bericht wird Angst Forschung beitragen.

Serie: Testing Animal Anxiety in Rats: Effects of Open Arm Ledges and Closed Arm Wall Transparency in Elevated Plus Maze Test

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

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 ...

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.

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 ...

The Double-H Maze: A Robust Behavioral Test for Learning and Memory in Rodents

Spatial cognition research in rodents typically employs the use of maze tasks, whose attributes vary from one maze to the next. These tasks vary by their behavioral flexibility and required memory duration, the number of goals and pathways, and also the overall task complexity. A confounding feature in many of these tasks is the lack of control over the strategy employed by the rodents to reach the goal, e.g., allocentric (declarative-like) or egocentric (procedural) based strategies. The double-H maze is a novel water-escape memory task that addresses this issue, by allowing the experimenter to direct the type of strategy learned during the training period. The double-H maze is a transparent device, which consists of a central alleyway with three arms protruding on both sides, along with an escape platform submerged at the extremity of one of these arms.
Rats can be trained using an allocentric strategy by alternating the start position in the maze in an unpredictable manner (see protocol 1; &#167;4.7), thus requiring them to learn the location of the platform based on the available allothetic cues. Alternatively, an egocentric learning strategy (protocol 2; &#167;4.8) can be employed by releasing the rats from the same position during each trial, until they learn the procedural pattern required to reach the goal. This task has been proven to allow for the formation of stable memory traces.
Memory can be probed following the training period in a misleading probe trial, in which the starting position for the rats alternates. Following an egocentric learning paradigm, rats typically resort to an allocentric-based strategy, but only when their initial view on the extra-maze cues differs markedly from their original position. This task is ideally suited to explore the effects of drugs/perturbations on allocentric/egocentric memory performance, as well as the interactions between these two memory systems.

The goal of this protocol is to investigate spatial cognition in rodents. The double-H water maze is a novel test, which is particularly useful to elucidate the different components of learning, consolidation and memory, as well as the interplay of memory systems.

Die Doppel-H Maze: A Robust Verhaltenstest für Lernen und Gedächtnis in Nagetiere

Spatial cognition research in rodents typically employs the use of maze tasks, whose attributes vary from one maze to the next. These tasks vary by their ...

Quantifying Cognitive Decrements Caused by Cranial Radiotherapy

With the exception of survival, cognitive impairment stemming from the clinical management of cancer is a major factor dictating therapeutic outcome. For many patients afflicted with CNS and non-CNS malignancies, radiotherapy and chemotherapy offer the best options for disease control. These treatments however come at a cost, and nearly all cancer survivors (~11 million in the US alone as of 2006) incur some risk for developing cognitive dysfunction, with the most severe cases found in patients subjected to cranial radiotherapy (~200,000/yr) for the control of primary and metastatic brain tumors1. Particularly problematic are pediatric cases, whose long-term survival plagued with marked cognitive decrements results in significant socioeconomic burdens2. To date, there are still no satisfactory solutions to this significant clinical problem.
We have addressed this serious health concern using transplanted stem cells to combat radiation-induced cognitive decline in athymic rats subjected to cranial irradiation3. Details of the stereotaxic irradiation and the in vitro culturing and transplantation of human neural stem cells (hNSCs) can be found in our companion paper (Acharya et al., JoVE reference). Following irradiation and transplantation surgery, rats are then assessed for changes in cognition, grafted cell survival and expression of differentiation-specific markers 1 and 4-months after irradiation. To critically evaluate the success or failure of any potential intervention designed to ameliorate radiation-induced cognitive sequelae, a rigorous series of quantitative cognitive tasks must be performed. To accomplish this, we subject our animals to a suite of cognitive testing paradigms including novel place recognition, water maze, elevated plus maze and fear conditioning, in order to quantify hippocampal and non-hippocampal learning and memory. We have demonstrated the utility of these tests for quantifying specific types of cognitive decrements in irradiated animals, and used them to show that animals engrafted with hNSCs exhibit significant improvements in cognitive function3.
The cognitive benefits derived from engrafted human stem cells suggest that similar strategies may one day provide much needed clinical recourse to cancer survivors suffering from impaired cognition. Accordingly, we have provided written and visual documentation of the critical steps used in our cognitive testing paradigms to facilitate the translation of our promising results into the clinic.

Cognitive impairment resulting from the radiotherapeutic management of brain tumors represents a clinically intractable condition that adversely impacts quality of life. The capability to critically evaluate potential interventions for ameliorating radiation-induced cognitive decrements ultimately depends on the capability to undertake rigorous quantitative assessments of cognition.

Quantifizierung Kognitive Dekrementiert von Cranial Strahlentherapie verursachten Schäden

With the exception of survival, cognitive impairment stemming from the clinical management of cancer is a major factor dictating therapeutic outcome. For ...

Medizin

Utilizing the Modified T-Maze to Assess Functional Memory Outcomes After Cardiac Arrest

Background: Evaluating mild to moderate cognitive impairment in a global cerebral ischemia (i.e. cardiac arrest) model can be difficult due to poor locomotion after surgery. For example, rats who undergo surgical procedures and are subjected to the Morris water maze may not be able to swim, thus voiding the experiment.
New Method: We established a modified behavioral spontaneous alternation T-maze test. The major advantage of the modified T-maze protocol is its relatively simple design that is powerful enough to assess functional learning/memory after ischemia. Additionally, the data analysis is simple and straightforward. We used the T-maze to determine the rats&#39; learning/memory deficits both in the presence or absence of mild to moderate (6 min) asphyxial cardiac arrest (ACA). Rats have a natural tendency for exploration and will explore the alternate arms in the T-maze, whereas hippocampal-lesioned rats tend to adopt a side-preference resulting in decreased spontaneous alternation ratios, revealing the hippocampal-related functional learning/memory in the presence or absence of ACA.
Results: ACA groups have higher side-preference ratios and lower alternations as compared to control.
Comparison with Existing Method(s): The Morris water and Barnes maze are more prominent for assessing learning/memory function. However, the Morris water maze is more stressful than other mazes. The Barnes maze is widely used to measure reference (long-term) memory, while ACA-induced neurocognitive deficits are more closely related to working (short-term) memory.
Conclusions: We have developed a simple, yet effective strategy to delineate working (short-term) memory via the T-maze in our global cerebral ischemia model (ACA).

This protocol describes the use of a modified T-maze to evaluate functional learning/memory in asphyxia cardiac arrest-induced cerebral ischemia.

Unter Verwendung der modifizierten T-Maze um Funktionsgedächtnis Ergebnisse nach Herzstillstand zu bewerten

Background: Evaluating mild to moderate cognitive impairment in a global cerebral ischemia (i.e. cardiac arrest) model can be difficult due to poor ...

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

Elevated Plus Maze Test Combined with Video Tracking Software to Investigate the Anxiolytic Effect of Exogenous Ketogenic Supplements

The overall goal of this study is to describe the methodology of the elevated plus maze (EPM) test in combination with a video tracking software. The purpose of the method is to document the effect of various potential anxiolytic treatments on laboratory rodent models. The EPM test is based on the rodents&#39; proclivity toward protected, enclosed dark spaces and unconditioned fear of open spaces and heights, and their innate intense motivation to explore novel environments. The EPM test is a widely used behavioral test for investigating the anxiolytic or anxiogenic responses of rodents given drugs that are known to affect behavior. Observation demonstrating a decreased proportion of time spent on closed arms, an increased proportion of time spent on open arms, a reduced number of entries to closed arms, and an elevated number of entries to open arms measured by the EPM test may reflect reduced anxiety levels. Using this method, the effect of exogenous ketone supplements on anxiety-related behavior is tested in Sprague Dawley (SPD) rats. Exogenous ketone supplements are chronically fed to the rats for 83 days or subchronically and acutely orally gavaged, daily for 7 days, before conducting the EPM test. Behavioral data collection is performed using the SMART video tracking system by a blinded observer at the end of the treatments. The main findings indicate that the EPM test is an effective method to detect the ketone supplement-induced anxiolytic effect and can be considered a sensitive measure to assess changes in anxiety behavior associated with drug- or metabolic-based therapies.

Here, we present a protocol to investigate changes in the anxiety level of rodent animal models. The elevated plus maze (EPM) test, used together with a video tracking software, provides a reliable method to document the effect of various potential anxiolytic treatments in preclinical laboratory scenarios.

Erhöhte Plus Labyrinth Test in Kombination mit Video Tracking-Software um die angstlösende Wirkung von exogenen ketogene Ergänzungen zu untersuchen

The overall goal of this study is to describe the methodology of the elevated plus maze (EPM) test in combination with a video tracking software. The ...

Utilizing a Reconfigurable Maze System to Enhance the Reproducibility of Spatial Navigation Tests in Rodents

Several maze shapes are used to test spatial navigation performance and behavioral phenotypes. Traditionally, each experiment requires a unique maze shape, thus requiring several separate mazes in different configurations. The maze geometry cannot be reconfigured in a single environment to accommodate scalability and reproducibility. The reconfigurable maze is a unique approach to address the limitations, allowing quick and flexible configurations of maze pathways in a repeatable manner. It consists of interlocking pathways and includes feeders, treadmills, movable walls, and shut-off sensors. The current protocol describes how the reconfigurable maze can replicate existing mazes, including the T-shaped, plus-shaped, W-shaped, and figure-eight mazes. Initially, the T-shaped maze was constructed inside a single experimental room, followed by modifications. The rapid and scalable protocol outlined herein demonstrates the flexibility of the reconfigurable maze, achieved through the addition of components and behavioral training phases in a stepwise manner. The reconfigurable maze systematically and precisely assesses the performance of multiple aspects of spatial navigation behavior.

The present protocol describes a reconfigurable maze, a unique system for testing spatial navigation and behavioral phenotypes in rodents. The adaptability of this maze system enables the execution of various experiments in a single physical environment. The ease of structural rearrangement generates reliable and reproducible experimental results.

Verwendung eines rekonfigurierbaren Labyrinthsystems zur Verbesserung der Reproduzierbarkeit von räumlichen Navigationstests bei Nagetieren

Several maze shapes are used to test spatial navigation performance and behavioral phenotypes. Traditionally, each experiment requires a unique maze ...

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 ...