The authors have no acknowledgements.

Ethics Statement: Alle Verfahren wurden von der Tierethikkommission der Universität von Sydney zugelassen und entsprach National Health und Medical Research Council of Australia Richtlinien. Die Verfahren wurden auf C57 / BL6J Mäusen, die an der Universität von Sydney Bosch Nagetier Einrichtung aufgezogen wurden durchgeführt. Alle Mäuse wurden in einem ausreichend belüfteten Raum bei 21 ° C Umgebungstemperatur auf einem 12 Stunden Licht-Dunkel-Zyklus wobei das Licht um 0600 h in einzeln belüfteten Käfigen mit freien Zugang zu Trockenfutter und Wasser untergebracht. Späte Schwangerschaft Frauen wurden randomisiert Standard- oder umwelt angereicherten Haltungsbedingungen zugeordnet. 1. Gehäuse (Enrichment Levels): <ol><li> Erhalten 4 späten Schwangerschaft erwachsenen weiblichen Mäusen. Zuordnet 2 auf die Standardbedingungen und legen Sie jeden von ihnen in ein sauberes Standardmaus Käfig (Abmessungen 391 x 199 x 160 mm), die eine rote Maus Iglu. Für die angereicherten Zustand, legen Sie die remaining 2-Mäuse in einem einzigen sauberen Ratte großen Käfig (Abmessungen 462 x 403 x 404 mm). </li><li> In den Käfig Bereicherung, legen eine Vielzahl von Objekten entwickelt, um sensorische und motorische Stimulation zu erhöhen (zB Laufräder, visuelle Reize, Duft Watte, Klettbänder). </li><li> Alle 2 bis 3 Tage verschieben diese Objekte über innerhalb des Käfigs; aktualisieren jede, die zerstört wurden. </li><li> Nach dem Absetzen nach 21 Tagen nach der Geburt, Geschlecht Tiere und Ort in die Mann-Frau-getrennt Gehäuse mit den ökologischen Zustand sie erhoben wurden. Aus dem angereicherten Zustand sicherzustellen, dass es zwischen 3 und 10 Mäuse pro Käfig mit 2 und 5 Mäuse für Standard. Beginnen Verhaltenstests einmal Tiere Erwachsenenalter (12-14 Wochen alt) zu erreichen. </li></ol> 2. Aufbau des Puzzlespiel-Kasten <ol><li> Erhalten 6 Stücke aus weißem Acryl (oder anderen nicht porösen Material): ein 750 x 280 mm, zwei 280 x 250 mm, zwei 750 x 250 mm und einer 150 x 280 mm (siehe 1). </li><li> Erhalten einem Stück schwarzem Acryl 280 x 250 mm, mit einer 40 x 40 mm Vierkantöffnung in einer Seite dieses Stück geschnitten (siehe Abbildung 1). </li><li> Montieren Sie die Puzzle Box wie folgt: Verwenden Sie den 750 x 280 mm Stück wie der Boden der Box, benutzen 280 x 250 mm Stücke wie die Enden der Box, und die 750 x 250 mm Stücke wie die Seiten der Box. </li><li> Messen Sie 150 mm in den Strafraum ein Ende, und legen Sie die schwarze Stück Acryl über den Kasten, so dass es spaltet sie in zwei Kammern (ein großer und ein kleiner) mit der Öffnung bündig mit der Unterseite der Box. </li><li> Nehmen Sie die 150 x 280 mm Stück weißes Acryl und legen Sie es oben auf dem kleineren Fach der Box sicherstellen, dass es deckt diesen Bereich komplett und bietet einen dunklen &quot;Ziel-box&quot; Kammer. Bringen Sie dieses Stück Acryl auf den Körper des Hauptfeld durch Scharniere, oder lassen Sie frei, während Verhaltenstests vollständig entfernt werden. </li><li> Nehmen Sie 3 Stücke aus Acryl (drei 4 x 120 mm) und kommen zu einem &quot;u-shap machened &quot;Kanal. </li></ol><img alt="Figur 1" fo:content-width="5in" src="/files/ftp_upload/52225/52225fig1highres.jpg" width="500" /> Abbildung 1: Schematische Darstellung des Puzzlespiel-Kasten Aufgabe. Das Puzzlespiel-Kasten ist ein Acryl-Box, bestehend aus einer offenen Feldbereich (600 x 280 mm) und einer geschützten Ziel-Feld-Bereich (150 x 280 mm), beträgt 750 x 280 mm in allen. Ein 40 x 40 mm Öffnung in der Trennwand (grau) die Trennung der beiden Bereiche erlaubt Tiere zur überdachten Ziel-Feld Bereich der offenen Feld zu gelangen. Diese Öffnung ist mit Hindernissen, die immer schwieriger zu entfernen, da Tests fort sind blockiert. Tiere werden einer Fünf-Tage-Protokoll, das aus vier Behinderung Bedingungen mit drei Studien für jede Bedingung. 3. Lauf des Puzzlespiel-Kasten Aufgabe <ol><li> Das Puzzle Box mit 70% Alkohol gründlich reinigen. Wiederholen Sie diesen Schritt zwischen jedem Tier getestet. </li><li> Ein sauberes rote Maus Iglu in der Ziel-box area des Puzzlespiel-Kasten, und ersetzen Sie den Deckel auf den Ziel-Box. </li><li> Wenn es ein Hindernis Zustand getestet werden, stellen Sie das Hindernis innerhalb der Tür des Ziel-Feld (siehe Abbildung 2). </li><li> Platzieren Sie die Maus, die am Ende am weitesten vom Ziel-Feld in die Freiflächen-Abschnitt des Puzzlespiel-Kasten, auf das Ziel-Feld ausgerichtet getestet und. </li><li> Notieren Sie sich die Zeit für alle vier Pfoten des Tieres getroffen werden, um das Ziel-Feld Teil des Puzzlespiel-Kasten geben. </li><li> Wenn ein Tier nicht in das Ziel-Feld, beenden Sie den Versuch, wenn die eingestellte Zeitlimit erreicht ist (siehe Tabelle 1). </li><li> Sobald eine Studie abgeschlossen ist, entfernen Sie das Tier aus der Puzzle-Box und legen Sie sie in einem separaten Haltekäfig, bis die nächste Verhandlung beginnt. Halten Sie eine 60 bis 180 sec Abstand zwischen Prüfungen für jedes Tier. </li><li> Für jedes Tier, führen drei Versuche pro Tag an fünf aufeinanderfolgenden Testtagen mit vier Behinderung Bedingungen und drei Versuche jeder Bedingung. Die dritte trial einer bestimmten Behinderung Bedingung sollte immer auf einer späteren Tag verabreicht (siehe Tabelle 1). </li></ol><img alt="Tabelle 1" fo:content-width="5in" src="/files/ftp_upload/52225/52225table1.jpg" width="500" /> Tabelle 1: Schema des Puzzlespiel-Kasten Aufgabe. Das Puzzle Box Aufgabe für fünf Tage laufen und besteht aus vier Behinderung Bedingungen. Es gibt drei Versuche auf jedem der ersten vier Tage, und man am fünften Tag. Jedes Hindernis Zustand hat drei Versuche; die ersten beiden an einem Tag, der dritte Tag unmittelbar nach. Der erste Versuch eines Hindernisses Zustand soll einheimische Problemlösungsfähigkeit zu testen, untersucht der zweite Prozess Aufgabe Erwerb und Verstärkung, und der dritte Versuch als Test zur Lösung Bindung und Rückruf verwendet. <img alt="Abbildung 2" fo:content-width="6in" src="/files/ftp_upload/52225/52225fig2highres.jpg" width="600" /> Abbildung 2: BehindernIonen-Bedingungen innerhalb des Puzzlespiel-Kasten Aufgabe. Schematische Darstellungen des Puzzlespiel-Kasten-Arena und der Behinderung Bedingungen in dieser Studie verwendet. (A) Bedingung 0 (C0), mit in der Tür zwischen der offenen Feld und Ziel-box Bereiche ohne Behinderung vorhanden. (B) Bedingung 1 (C1 ), mit einem &quot;u förmigen&quot; vorhanden Kanal innerhalb der Tür zwischen der offenen Feld und Ziel-box Bereichen. (C) Bedingung 2 (C2), in dem der Kanal mit saubere Bettwäsche Material gefüllt ist. (D) Bedingung 3 (C3 ), mit einem &quot;Gewebe-Plug&quot; in der Tür zwischen der offenen Feld und Ziel-box Bereichen vorhanden. (E) Bedingung 4 (C4) mit einem &quot;Schaum-Stecker&quot; in der Tür zwischen der offenen Feld und Ziel-Feld vorhanden Bereichen. Abmessungen der Arena sind nach Methoden und in Abbildung 1 aufgeführt. 4. Auswertung der Daten <ol><li> Stellen Sie Daten entweder als Zeit genommen comrunden das Test (einschließlich Null-Studien, in denen Tiere nicht die Aufgabe, innerhalb der festgelegten Frist zu vervollständigen), oder als die Anzahl der Null-Studien. </li><li> Verwenden Sie eine ANOVA mit wiederholten Messungen, um die Wirkung der Wohnanlage auf Leistung innerhalb des Puzzlespiel-Kasten zu bewerten, mit Behinderung Art / Zustand (C) und die Task-Nummer (T) Innersubjektfaktoren und Anreicherungsgrad (Standard gegenüber angereichertem) als Zwischensubjektfaktor . </li></ol>

The authors declare they have no competing financial interests.

Die vorgelegten Daten zeigen, dass das Puzzle Box kann effektiv verwendet werden, um die Auswirkungen von EE zu beurteilen. Mäuse in angereicherten Umgebungen erhöht konsequent nahmen deutlich weniger Zeit zum Hindernis Rätsel in diesem Verhaltenstest als getan Tiere in Standard-Laborbedingungen erhoben lösen. Darüber hinaus war dieser Unterschied am deutlichsten in der ersten Studie für drei der vier getesteten Bedingungen, was darauf hindeutet, EE hat einen größeren Einfluss auf die einheimische Problemlösung...

Umweltanreicherung (EE) als Umgebung, die Tiere mit einer erhöhten Chance für die soziale Interaktion, Motorik und größere sensorische Stimulation als normalerweise in einem Standard-Labor environment.EE erlebt bieten hat gezeigt, dass durchgängig auf das Verhalten der Tiere, was zu definieren Veränderungen wie Reduktion von Stress und Angst im Zusammenhang mit Aktivitäten 8-10, verbesserte Leistung bei Lern- und Gedächtnisaufgaben 8,11, frühe Beginn der motorischen Koordination und explorative Aktivität 11, Änderungen der mütterlichen Fürsorge 8 sowie Beständigkeit gegenüber Suchtmitteln 12- 15. Ferner EE offenbart worden, um die Auswirkungen von neurodegenerativen Erkrankungen zu lindern, Verzögerung des Einsetzens und Verringern der Schwere der Symptome im Tiermodell der Huntington-1-4,16, Parkinson-17 und Alzheimer 18. Diese Veränderungen corbetreffen den anatomischen und molekularen Veränderungen EE ist bekannt, im Gehirn zu induzieren. Tiere in angereicherten Umgebungen von frühen Stadien der Entwicklung erhöht zeigen eine Vielzahl von neuronalen Veränderungen, einschließlich der erhöhten Hirngewicht und Rindendicke, dendritischen Verzweigungs 9,2-22 und Synapsendichte 23. EE kann sowohl die Höhe und den Zeitpunkt der Wachstumsfaktor Ausdruck 9,24-30, die gezeigt wurde, um zu einer beschleunigten Entwicklung von sensorischen 25,26,28,29, mnemonic 30 sowie Motorschaltungen 31,32 beitragen verändern. Frühere Arbeiten haben manchmal widersprüchliche Ergebnisse zeigten bei der Untersuchung der Auswirkungen von EE, ohne Rücksicht auf die verschiedenen Arten von Tieren und die Umwelt in den einzelnen Studien 9,24,27,30 benutzt. Zur Zeit gibt es keine einheitliche und einfache Verhaltens Aufgabe, die verwendet werden können, um die Wirksamkeit der verschiedenen EE Paradigmen in verschiedenen Stämmen und spec messener Jahren von Tieren. Das Puzzle Box Aufgabe wurde als einfacher Test, um festzustellen Mutter Problem eines Tieres Lösungsfähigkeit 7 konzipiert. Tiere in den offenen Bereich platziert sind erforderlich, um zu entfernen behindern Vormaterialien in eine kleine Öffnung, um einen überdachten Bereich / Unterkünfte zugreifen entfernt. Jedes Thema wird von drei Studien mit der gleichen Behinderung, um die drei verschiedenen kognitiven Eigenschaften beurteilen gegeben. Die erste Studie liefert einen Ausgangswert Anzeige des inhärenten oder native Problemlösungsfähigkeit. Die zweite Studie, am selben Tag ausgeführt wird, gibt einen Hinweis auf die Fähigkeit des Tieres, auf zu verbessern und somit für eine verstärke Strategien zur Entfernung des spezifischen Behinderung. Der dritte Versuch, am folgenden Tag durchgeführt wurde, gibt einen Einblick in die Fähigkeit des Subjekts zu behalten und erinnern an die gelernt Lösung der Aufgabe. Die Motivation für die Lösung dieser &quot;Obstruktion Puzzles&quot; der Tiere können variiert werden, potentiellerinnert an eine angeborene Wunsch, offene Felder zu vermeiden und suchen Schutz, sowie einen inhärenten Antrieb zu ihrer Umgebung 6,7 erkunden. Die Vielzahl möglicher Verhaltens Fahrer den Wunsch, das Puzzle zu lösen Box zugrunde legt nahe, dass verschiedene Bereiche des Gehirns sind bei der Vermittlung Aufgabenwahrnehmung beteiligt. Frühere Arbeiten haben gezeigt, dass in murinen Modellen von Schizophrenie, der präfrontalen Hirnrinde und der Hippocampus werden in der Übernahme dieser Aufgabe 5 beteiligt. Eine Läsion Studie an Ratten wurde auch gezeigt, eine große Anzahl von Gehirnbereichen in Puzzle Box Leistung, einschließlich verschiedener Thalamus, Hypothalamus, Kleinhirn und limbischen Strukturen beteiligt. Zusammen zeigen diese Ergebnisse, dass die Durchführung dieser Problemlösung Aufgabe umfasst eine Vielzahl von Nervenstrukturen mit kognitiven Funktion assoziiert. Das Puzzle Box wurde erfolgreich verwendet, um die Problemlösungsfähigkeit von Mäusen sowie kognitive Defizite durch m ausgestellt bewertenUrin-Modelle der Schizophrenie 5-7. Leistung auf der Aufgabe hat sich gezeigt, dass sehr konsistente und gut mit Ergebnissen von anderen kognitiven Verhaltenstests 6 korrelieren. Das Ziel dieser Arbeit war es daher, die Puzzlespiel-Kasten Aufgabe anzupassen, eine einfache und zuverlässige Mittel zur Bestimmung der Wirksamkeit der EE werden.

<table><tbody><tr><td>Black Acrylic Board</td><td></td><td></td><td>280 x 250 mm with a 40 x 40 mm &quot;door&quot; cut into one side</td></tr><tr><td>White Acrylic Board</td><td></td><td></td><td>280 x 250 mm</td></tr><tr><td>White Acrylic Board</td><td></td><td></td><td>280 x 250 mm</td></tr><tr><td>White Acrylic Board</td><td></td><td></td><td>750 x 250 mm</td></tr><tr><td>White Acrylic Board</td><td></td><td></td><td>750 x 250 mm</td></tr><tr><td>White Acrylic Board</td><td></td><td></td><td>150 x 280 mm</td></tr><tr><td>White Acrylic Board</td><td></td><td></td><td></td></tr><tr><td>Underpass</td><td></td><td></td><td>3 pieces of 40 x 120 mm plexiglass or acrylic</td></tr><tr><td colspan="4">Note: If unable to access acrylic board, plexiglass or similar non-porous material will suffice.</td></tr><tr><td>Webcam</td><td>Logitech</td><td>C210</td><td>Fix to roof with electrical tape. Alternatively, use a tripod.</td></tr><tr><td>VirtualDub v1.10.4</td><td>VirtualDub</td><td>N/A&amp;nbsp;</td><td>Software for recording behaviour videos. Input from webcam.</td></tr><tr><td>TopScan v 3.0</td><td>CleverSys Inc.</td><td>N/A&amp;nbsp;</td><td>Software for automated top-view tracking and analysis of mouse behaviour. Captured videos are analysed post-hoc.</td></tr></tbody></table>

the use of the puzzle box as a means of assessing the efficacy of environmental enrichment

Environmental enrichment can dramatically influence the development and function of neural circuits. Further, enrichment has been shown to successfully delay the onset of symptoms in models of Huntington&#8217;s disease 1-4, suggesting environmental factors can evoke a neuroprotective effect against the progressive, cellular level damage observed in neurodegenerative disorders. The ways in which an animal can be environmentally enriched, however, can vary considerably. Further, there is no straightforward manner in which the effects of environmental enrichment can be assessed: most methods require either fairly complicated behavioral paradigms and/or postmortem anatomical/physiological analyses. This protocol describes the use of a simple and inexpensive behavioral assay, the Puzzle Box 5-7 as a robust means of determining the efficacy of increased social, sensory and motor stimulation on mice compared to cohorts raised in standard laboratory conditions. This simple problem solving task takes advantage of a rodent&#8217;s innate desire to avoid open enclosures by seeking shelter. Cognitive ability is assessed by adding increasingly complex impediments to the shelter&#8217;s entrance. The time a given subject takes to successfully remove the obstructions and enter the shelter serves as the primary metric for task performance. This method could provide a reliable means of rapidly assessing the efficacy of different enrichment protocols on cognitive function, thus paving the way for systematically determining the role specific environmental factors play in delaying the onset of neurodevelopmental and neurodegenerative disease.

Die hier beschriebenen Ergebnisse sind eine repräsentative Stichprobe, mit Daten aus mehreren Jahrgängen aus verschiedenen Würfen übernommen. Alle Verhaltenstests wurde zwischen 0700 und 1100 Stunden durchgeführt, wobei randomisierte Prüfung, um von Tieren innerhalb einer Kohorte. Tiere in einer angereicherten Umwelt erhöht (n = 14, 7 weibliche und 7 männliche) nahmen deutlich weniger Zeit, um die Behinderung Aufgaben innerhalb des Puzzlespiel-Kasten als die innerhalb einer Standardumgebung erhöht lösen (n = 1...

Watch this Scientific Journal Video about The Use of the Puzzle Box as a Means of Assessing the Efficacy of Environmental Enrichment at JoVE.com

The Use of the Puzzle Box as a Means of Assessing the Efficacy of Environmental Enrichment

Environmental enrichment provides a potential protective effect against neurodegenerative disorders. Currently, however, there is no easy way of determining the efficacy of enrichment procedures. This protocol describes a simple &#8220;Puzzle Box&#8221; method for assessing an animal&#8217;s cognitive function, in order to reveal the effectiveness of environmental enrichment.

Die Nutzung des Puzzlespiel-Kasten als Mittel zur Bewertung der Wirksamkeit von Umwelt Enrichment

Environmental enrichment can dramatically influence the development and function of neural circuits.  Further, enrichment has been shown to successfully ...

the-use-puzzle-box-as-means-assessing-efficacy-environmental

Research

JoVE Journal

Behavior

11.6K Aufrufe.  University of Sydney. Environmental enrichment provides a potential protective effect against neurodegenerative disorders. Currently, however, there is no easy way of determining the efficacy of enrichment procedures. This protocol describes a simple “Puzzle Box” method for assessing an animal’s cognitive function, in order to reveal the effectiveness of environmental enrichment.

Serie: The Use of the Puzzle Box as a Means of Assessing the Efficacy of Environmental Enrichment

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

This protocol describes the modified hole board (mHB), which combines features from a traditional hole board and open field and is designed to measure multiple dimensions of unconditioned behavior in small laboratory mammals (e.g., mice, rats, tree shrews and small primates). This paradigm is a valuable alternative for the use of a behavioral test battery, since a broad behavioral spectrum of an animal&#8217;s behavioral profile can be investigated in one single test.
The apparatus consists of a box, representing the &#8216;protected&#8217; area, separated from a group compartment. A board, on which small cylinders are staggered in three lines, is placed in the center of the box, representing the &#8216;unprotected&#8217; area of the set-up. The cognitive abilities of the animals can be measured by baiting some cylinders on the board and measuring the working and reference memory. Other unconditioned behavior, such as activity-related-, anxiety-related- and social behavior, can be observed using this paradigm. Behavioral flexibility and the ability to habituate to a novel environment can additionally be observed by subjecting the animals to multiple trials in the mHB, revealing insight into the animals&#8217; adaptive capacities.
Due to testing order effects in a behavioral test battery, na&#239;ve animals should be used for each individual experiment. By testing multiple behavioral dimensions in a single paradigm and thereby circumventing this issue, the number of experimental animals used is reduced. Furthermore, by avoiding social isolation during testing and without the need to food deprive the animals, the mHB represents a behavioral test system, inducing if any, very low amount of stress.

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.

Der modifizierte Loch Board - Mess Verhalten, Kognition und Sozialinteraktion an Mäusen und Ratten

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

Forschung

Verhalten

A Protocol for Housing Mice in an Enriched Environment

Environmental enrichment (EE) is a housing environment for mice that boosts mental and physical health compared to standard laboratory housing. Our recent studies demonstrate that environmental enrichment decreases adiposity, increases energy expenditure, resists diet induced obesity, and causes cancer remission and inhibition in mice. EE typically consists of larger living space, a variety of &#8216;toys&#8217; to interact with, running wheels, and can include a number of other novel environmental changes. All of this fosters a more complex social engagement, cognitive and physical stimulations. Importantly, the toy location and type of toy is changed regularly, which encourages the mice to adapt to a frequently changing and novel environment. Many variables can be manipulated in EE to promote health effects in mice. Thus these approaches are difficult to control and must be properly managed to successfully replicate the associated phenotypes. Therefore, the goal of this video is to demonstrate how EE is properly set up and maintained to assure a complex, challenging, and controlled environment so that other researchers can easily reproduce the protective effects of EE against obesity and cancer.

Environmental enrichment for mice requires a complex and challenging setup, as well as comprehensive husbandry and handling techniques to assure robust metabolic and anti-cancer effects in the mice. This protocol provides detailed procedures to reproduce the above mentioned effects in mice.

Ein Protokoll für die Housing Mäuse in einer angereicherten Umwelt

Environmental enrichment (EE) is a housing environment for mice that boosts mental and physical health compared to standard laboratory housing. Our recent ...

Assessing Burrowing, Nest Construction, and Hoarding in Mice

Deterioration in the ability to perform "Activities of daily living" (ADL) is an early sign of Alzheimer's disease (AD). Preclinical behavioural screening of possible treatments for AD currently largely focuses on cognitive testing, which frequently demands expensive equipment and lots of experimenter time. However, human episodic memory (the most severely affected aspect of memory in AD) is different to rodent memory, which seems to be largely non-episodic. Therefore the present ways of screening for new AD treatments for AD in rodents are intrinsically unlikely to succeed. A new approach to preclinical screening would be to characterise the ADL of mice. Fortuitously, several such assays have recently been developed at Oxford, and here the three most sensitive and well-characterised are presented.
Burrowing was first developed in Oxford13. It evolved from a need to develop a mouse hoarding paradigm. Most published rodent hoarding paradigms required a distant food source to be linked to the home cage by a connecting passage. This would involve modifying the home cage as well as making a mouse-proof connecting passage and food source. So it was considered whether it would be possible to put the food source inside the cage. It was found that if a container was placed on the floor it was emptied by the next morning., The food pellets were, however, simply deposited in a heap at the container entrance, rather than placed in a discrete place away from the container, as might be expected if the mice were truly hoarding them. Close inspection showed that the mice were performing digging ("burrowing") movements, not carrying the pellets in their mouths to a selected place as they would if truly hoarding them.6



Food pellets are not an essential substrate for burrowing; mice will empty tubes filled with sand, gravel, even soiled bedding from their own cage. Moreover, they will empty a full tube even if an empty one is placed next to it8.



Several nesting protocols exist in the literature. The present Oxford one simplifies the procedure and has a well-defined scoring system for nest quality5. 



A hoarding paradigm was later developed in which the mice, rather than hoarding back to the real home cage, were adapted to living in the "home base" of a hoarding apparatus. This home base was connected to a tube made of wire mesh, the distal end of which contained the food source. This arrangement proved to yield good hoarding behaviour, as long as the mice were adapted to living in the "home base" during the day and only allowed to enter the hoarding tube at night.

Burrowing, nesting, and hoarding are species-typical activities that mice readily perform in the laboratory. This article describes how they can be easily and cheaply assessed. These protocols are extremely sensitive to mouse strain, brain lesions and diseases. Moreover they constitute &#x201C;environmental enrichment&#x201D; for the mice, and embody the &#x201C;Refinement&#x201D; aspect of the &#x201C;3 Rs&#x201D;18.

Die Beurteilung Burrowing, Nest Bau und Horten in Mäuse

Deterioration in the ability to perform "Activities of daily living" (ADL) is an early sign of Alzheimer's disease (AD). Preclinical behavioural screening ...

Neurowissenschaften

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 ...

Introducing Clicker Training as a Cognitive Enrichment for Laboratory Mice

Establishing new refinement strategies in laboratory animal science is a central goal in fulfilling the requirements of Directive 2010/63/EU. Previous research determined a profound impact of gentle handling protocols on the well-being of laboratory mice. By introducing clicker training to the keeping of mice, not only do we promote the amicable treatment of mice, but we also enable them to experience cognitive enrichment. Clicker training is a form of positive reinforcement training using a conditioned secondary reinforcer, the "click" sound of a clicker, which serves as a time bridge between the strengthened behavior and an upcoming reward. The effective implementation of the clicker training protocol with a cohort of 12 BALB/c inbred mice of each sex proved to be uncomplicated. The mice learned rather quickly when challenged with tasks of the clicker training protocol, and almost all trained mice overcame the challenges they were given (100% of female mice and 83% of male mice). This study has identified that clicker training for mice strongly correlates with reduced fear in the mice during human-mice interactions, as shown by reduced anxiety-related behaviors (e.g., defecation, vocalization, and urination) and fewer depression-like behaviors (e.g., floating). By developing a reliable protocol that can be easily integrated into the daily routine of the keeping of laboratory mice, the lifetime experience of welfare in the mice can be improved substantially.

The development of new refinement strategies for laboratory mice is a challenging task that contributes towards fulfilling the 3R principle. This protocol introduces clicker training as a cognitive enrichment program for laboratory mice.

Einführung in Clicker Ausbildung als Cognitive Anreicherung für Labormäuse

Establishing new refinement strategies in laboratory animal science is a central goal in fulfilling the requirements of Directive 2010/63/EU. Previous ...

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. ...

Automated, Long-term Behavioral Assay for Cognitive Functions in Multiple Genetic Models of Alzheimer's Disease, Using IntelliCage

Multiple factors&#8212;such as aging and genes&#8212;are frequently associated with cognitive decline. Genetically modified mouse models of cognitive decline, such as Alzheimer&#39;s disease (AD), have become a promising tool to elucidate the underlying mechanisms and promote the therapeutic advances. An important step is the validation and characterization of expected behavioral abnormality in the models, in the case of AD, cognitive decline. The long-term behavioral investigations of laboratory animals to study the effect of aging demand substantial efforts from researchers. The IntelliCage system is a high-throughput and cost-effective test battery for mice that eliminates the need for daily human handling. Here, we describe how the system is utilized in the long-term phenotyping of a genetic Alzheimer&#39;s disease model, specifically focusing on the cognitive functions. The experiment employs repeated battery of tests that assess spatial learning and executive functions. This cost-effective age-dependent phenotyping allows us to identify the transient and/or permanent effects of genes on various cognitive aspects.

This paper describes a protocol for cognitive assessments for genetic models of the Alzheimer&#39;s disease using the IntelliCage system, which is a high throughput automated behavioral monitoring system with operant conditioning.

Automatisiert, langfristige Verhaltensstörungen Assay für kognitive Funktionen in mehreren genetischen Modellen der Alzheimer-Krankheit, mit IntelliCage

Multiple factors&#8212;such as aging and genes&#8212;are frequently associated with cognitive decline. Genetically modified mouse models of cognitive ...

Simultaneous Monitoring of Wireless Electrophysiology and Memory Behavioral Test as a Tool to Study Hippocampal Neurogenesis

Brainwaves amplitude obtained from electroencephalography (EEG) has been well-recognized as a basis for cognitive capacity, memory, and learning on animals and humans. Adult neurogenesis mechanism is also linked to memory and learning improvement. Traditionally, researchers used to assess learning and memory parameters in rodent models by behavioral tasks. Therefore, the simultaneous monitoring of behavioral changes and EEG is particularly interesting in correlating data between brain activity and task-related behaviors. However, most of the equipment required to perform both studies are either complex, expensive, or uses a wired setup network that hinders the natural animals&#8217; movement. In this study, EEG was recorded with a wireless electrophysiology device along with the execution of a novel object recognition task (NORT). The animal&#8217;s behavior was monitored simultaneously by a video tracking system. Both recordings were analyzed offline by their timestamps which were synched to link EEG signals with the animal&#8217;s actions. Subjects consist of adult Wistar rats after medium-term environmental enrichment treatment. Six skull screw electrodes were fixed in pairs on both hemispheres over frontal, central, and parietal regions and were referenced to an electrode located posterior of the nasal bone. NORT protocol consists of exposing the animal to two identical objects for 10 min. After 2 h and 24 h, one of the objects was replaced with a novel one. Exploration time for each object was monitored by a behavioral tracking software (BTS) and EEG data recording. The analysis of the EEG synced with behavioral data consists of estimations of alpha and beta relative band power and comparisons between novel object recognition versus familiar object exploration, between three experimental stages. In this manuscript, we have discussed electrodes manufacturing process, epidural electrodes implantation surgery, environmental enrichment protocol, NORT protocol, BTS setup, EEG &#8211; BTS coupling for simultaneous monitoring in real-time, and EEG data analysis based on automatic events detection.

The protocol presented here provides information on the simultaneous electroencephalography (EEG) and behavioral assessment in real-time. We have discussed all steps involved in this protocol as an attractive solution for researchers in many fields of neuroscience, particularly in learning and memory areas.

Simultane Überwachung der drahtlosen Elektrophysiologie und des Gedächtnisverhaltenstests als Werkzeug zur Untersuchung der Neurogenese des Hippocampus

Brainwaves amplitude obtained from electroencephalography (EEG) has been well-recognized as a basis for cognitive capacity, memory, and learning on ...

The Innovation Arena: A Method for Comparing Innovative Problem-Solving Across Groups

Problem-solving tasks are commonly used to investigate technical, innovative behavior but a comparison of this ability across a broad range of species is a challenging undertaking. Specific predispositions, such as the morphological toolkit of a species or exploration techniques, can substantially influence performance in such tasks, which makes direct comparisons difficult. The method presented here was developed to be more robust with regard to such species-specific differences: the Innovation Arena presents 20 different problem-solving tasks. All tasks are presented simultaneously. Subjects are confronted with the apparatus repeatedly, which allows a measurement of the emergence of innovations over time - an important next step for investigating how animals can adapt to changing environmental conditions through innovative behavior.
Each individual was tested with the apparatus until it ceased to discover solutions. After testing was concluded, we analyzed the video recordings and coded successful retrieval of rewards and multiple apparatus-directed behaviors. The latter were analyzed using a Principal Component Analysis and the resulting components were then included in a Generalized Linear Mixed Model together with session number and the group comparison of interest to predict the probability of success.
We used this approach in a first study to target the question of whether long-term captivity influences the problem-solving ability of a parrot species known for its innovative behavior: the Goffin&#180;s cockatoo. We found an effect in degree of motivation but no difference in the problem-solving ability between short- and long-term captive groups.

The Innovation Arena is a novel comparative method for studying technical innovation rate per time unit in animals. It is composed of 20 different problem-solving tasks, which are presented simultaneously. Innovations can be carried out freely and the setup is robust with regard to predispositions on an individual, population or species level.

Die Innovationsarena: Eine Methode zum Vergleich innovativer Problemlösungen zwischen Gruppen

Problem-solving tasks are commonly used to investigate technical, innovative behavior but a comparison of this ability across a broad range of species is ...

Assessment of Mouse Judgment Bias through an Olfactory Digging Task

Judgment biases (JB) are differences in the way that individuals in positive and negative affective/emotional states interpret ambiguous information. This phenomenon has long been observed in humans, with individuals in positive states responding to ambiguity 'optimistically' and those in negative states instead showing 'pessimism'. Researchers aiming to assess animal affect have taken advantage of these differential responses, developing tasks to assess judgment bias as an indicator of affective state. These tasks are becoming increasingly popular across diverse species and fields of research. However, for laboratory mice, the most widely used vertebrates in research and a species heavily relied upon to model affective disorders, only one JB task has been successfully validated as sensitive to changes in affective state. Here, we provide a detailed description of this novel murine JB task, and evidence of its sensitivity to mouse affect. Though refinements are still necessary, assessment of mouse JB opens the door for answering both practical questions regarding mouse welfare, and fundamental questions about the impact of affective state in translational research.

This article provides a detailed description of a novel mouse judgment bias protocol. Evidence of this olfactory digging task's sensitivity to affective state is also demonstrated and its utility across diverse research fields is discussed.

Bewertung der Mausurteilsverzerrung durch eine olfaktorische Grabaufgabe

Judgment biases (JB) are differences in the way that individuals in positive and negative affective/emotional states interpret ambiguous information. This ...