This work was supported by the Translational Animal Research Center of the University Medical Center of the Johannes Gutenberg-University Mainz. The authors are most grateful to Thomas Wacker for his technical support.

Ethik-Statement: Die Handhabung der Mäuse und die experimentellen Verfahren wurden in Übereinstimmung mit den europäischen, nationalen und institutionellen Richtlinien für die Tierpflege durchgeführt. HINWEIS: Das Protokoll enthält fünf Tagen Interventionen (Montag bis Freitag), mit Pausen an den Wochenenden (Samstag und Sonntag). Das Protokoll kann leicht zu erfüllen spezifischen Bedürfnisse angepasst werden. 1. Festlegung einer Belohnung geeignet als Zweit Reinforcer HINWEIS: Verwenden Sie Lebensmittel Belohnungen, wie vakuumverpackt Lebens- oder Futtermitteln, die Lebensmittelsicherheitsstandards erfüllt. Zum Beispiel können verschiedene Arten von Nüssen, Schokolade, Gummibärchen oder getrocknete Früchte sind geeignet. <ol><li> Legen Sie eine kleine Petrischale mit verschiedenen Lebensmitteln Belohnungen (Stücke von 0,5 cm 3) in den Heimkäfig (Abbildung 1). </li><li> Nach 10 min prüfen Reste. </li><li> Wenn alle Belohnungen nach 10 Minuten gegessen werden, verkürzen die Zeitspanne, bis eine Präferenz geschätzt werden kann. </li><li> An den Tagen 4 und 5 (zB Thursday und Freitag), fügen Sie kleine Mengen des bevorzugten ( &quot;Gewinner&quot;) Belohnung für die Heimat-Käfig eine Affektiertheit aller Mäuse im Käfig auf die Belohnung zu etablieren. </li><li> Wenn die Mäuse auf Maus - Tunnel nicht gewöhnte werden, fügen Sie einen Tunnel zu dem Käfig (mindestens 2 Tage vor dem Beginn der Ausbildung, zum Beispiel über das Wochenende). </li><li> Wiederholen Sie die Schritte 1,2 bis 1,4 auf 3 aufeinanderfolgenden Tagen (zB Montag bis Mittwoch). </li></ol> 2. Clicker Trainings <ol><li> Die Vorbereitungen für jede Trainingseinheit HINWEIS: Jedes Mal, wenn eine Maus in diesem Protokoll werden muss angehoben, heben Sie ihn in einer ruhigen und sanften Art und Weise. Legen Sie eine Hand unter die Maus, um es zu heben. Während der ersten paar Mal, könnte die Mäuse gerührt werden, aber die Gewöhnung Prozess wird nach einigen Tagen beendet sein. Wenn dieser Schritt Schwierigkeiten bereitet , ist es nützlich , für &quot;Schröpfen auf der offenen Seite&quot; 28 zu dem Protokoll zu bezeichnen. <ol><li> Bereiten Sie einen Käfig mit Einbettungsmaterial. </li><li> Transfer der Käfig der Maus an einen ruhigen Ort ausgebildet. </li><li> Bereiten Sie die Belohnung und haben einen Timer und den Clicker / Ziel-Stick Kombination bereit. HINWEIS: Wenn die Mäuse auf die Handhabung gewöhnt sind und haben bereits ein Vertrauensverhältnis mit dem Experimentator, können die Mäuse mit der Hand gefüttert werden. Andernfalls fügen Sie die Belohnung an einem Stock oder einer Zange in der Weise, dass die Mäuse einen Abstand von dem Experimentator zu halten. </li><li> Entfernen Sie alle Objekte aus dem Hause Käfig, die nun als Trainingsbereich dient (zB Maus Häuser, Nestbaumaterial usw.). </li><li> Übertragen Sie die Käfig Begleiter zu einem vorbereiteten Käfig. </li></ol></li><li> Allgemeine Regeln, die für alle Trainingseinheiten gelten HINWEIS: Die folgenden allgemeinen Regeln gelten für alle Sitzungen und werden nicht im gesamten Protokoll wieder erwähnt werden. <ol><li> Trainieren Sie jede Maus für 5 min. Wechseln Sie zwischen 30-s Ausbildung und 15-s Pausenzeiten. </li><li> Lassen Sie die Maus auf die Belohnung nagen nicht mehr als eine Sekunde. Dann nehmen Sie den Stick (mit der Belohnung) aus dem Käfig. </li><li> Führen Sie das folgende Muster zu belohnen: 1 - 10 Aufführungen von Verhalten: jedes Mal belohnen. 11-21 Leistung des Verhaltens: jedes zweite Mal belohnen. Von 22 Performance des Verhaltens auf: jeden dritten Mal belohnen. </li><li> Nach 4 min der Ausbildung, wird die nächste Leistung des gewünschten Verhaltens durch einen Jackpot Belohnung verstärkt. Lassen Sie die Maus auf die Futterbelohnung dreimal länger als bisher zu nagen. </li></ol></li><li> Training Tag 1: &quot;Die Verknüpfung der sekundären reinforcer mit dem Futterbelohnung&quot; <ol><li> Stellen Sie den Timer auf 5 Minuten, Start-Taste drücken, und fügen Sie die Maus-Tunnel zum Heimkäfig. HINWEIS: Nutzen Sie die angeborene Thigmotaxis von Mäusen und platzieren Sie den Tunnel an einer Wand (Abbildung 2). Für die ersten paar Sitzungen, dies wird die Wahrscheinlichkeit, dass der Maus-Eingabe der Tunnel zu erhöhen. </li><li> Warten Sie, bis die Maus, um den Tunnel kontrolliert. Sobald der mouse betritt den Tunnel, und klicken Sie auf die Belohnung am Ende des Tunnels präsentieren. </li><li> Lassen Sie die Maus-Feed auf die Belohnung, während im Tunnel sitzen. </li><li> Klicken Sie kontinuierlich für 15 s, während die Maus im Tunnel sitzt. </li><li> Sobald die Maus, um den Tunnel verlässt, gehen Sie zurück zu Schritt 3.2. </li></ol></li><li> Training Tage 2 bis 5: &quot;Laufen durch einen Tunnel&quot; <ol><li> Stellen Sie den Timer auf 5 Minuten, Start-Taste drücken, und fügen Sie die Maus-Tunnel zum Heimkäfig. </li><li> Sobald die Maus in den Tunnel einfährt, klicken und die Belohnung am Ende des Tunnels präsentieren. </li><li> Solange die Maus im Tunnel ist, klicken und sofort die Belohnung in der gleichen Art und Weise präsentieren. </li><li> Wiederholen Sie diesen Vorgang für die nächsten 30 s, und dann Pause für 15 s in den Tunnel aus dem Käfig zu nehmen. </li><li> Fügen Sie den Tunnel wieder und sofort klicken, wenn die Maus den Tunnel wieder eintritt. </li><li> Präsentieren Sie die Belohnung am Ende des Tunnels. </li><li> Lassen Sie die Maus für bis zu 1 s zu nagen. </li&gt; <li> Nehmen Sie die Belohnung weg. </li><li> Sobald die Maus beginnt , indem sich in den Tunnel Wiedereintritt, präsentieren die Belohnung vor dem Ende des Tunnels (3). </li></ol></li><li> Training Woche 2: &quot;Nach einem Ziel-Stick&quot; <ol><li> Stellen Sie den Timer auf 5 Minuten, Start-Taste drücken, und legen Sie die Kugel am Ende des Ziel-Stick in den Käfig. </li><li> Sobald die Maus Interesse in der ganzen Welt zeigt, klicken und die Belohnung neben dem Globus (4 und 5) präsentieren. </li><li> Belohnen Sie die Maus erst, nachdem es um den Globus mit seiner Nase berührt. </li><li> Wenn die Maus , um die durchgeführte Aktion zur Belohnung verknüpft ist, ändern Sie die Position der Kugel während der Trainingseinheit (Abbildung 6). </li><li> Warten Sie auf die Maus, um dieses letzte Herausforderung zu begegnen. </li><li> Legen Sie die Kugel in den Käfig. </li><li> Kurz bevor die Maus, um den Globus berührt, langsam und vorsichtig die Position der Kugel von 1 cm ändern. </li&gt; <li> Klicken und belohnen, wenn die Maus den Abstand und berührt den Globus überschritten hat. </li><li> Wiederholen Sie diesen Vorgang, bis die Maus zuverlässig folgt. </li><li> erstrecken sich langsam den Abstand der Maus zu überqueren hat. </li></ol></li><li> Training Woche 3: &quot;Nach dem Ziel-Stick an den Experimentator Hand&quot; <ol><li> Legen Sie eine Hand in der Trainings Käfig während der Clicker / Ziel-Stick Kombination halten und die Belohnung mit der anderen Hand. </li><li> Stellen Sie den Timer auf 5 Minuten, Start-Taste drücken, und legen Sie die Kugel am Ende des Zielstock neben der Hand. </li><li> Sobald die Maus Interesse in der ganzen Welt zeigt, klicken und die Belohnung neben dem Globus präsentieren. </li><li> Sobald die Maus sich dieser Herausforderung gestellt hat, bewegen Sie die Kugel mehrere Schritte näher an der Hand. </li><li> Legen Sie die Kugel auf der Handfläche der Hand. Klicken und belohnen , während die Maus auf der Handfläche der Hand sitzt (Abbildung 7). </li></ol></li></ol>

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The authors have nothing to disclose.

Die effektive Umsetzung des Clicker-Training-Protokoll mit einer Kohorte von 12 Balb / c-Inzuchtmäusen beiderlei Geschlechts erwies sich als unkompliziert zu sein. Frühere Studien haben die Wirksamkeit von Clickertraining mit mehreren Arten bestätigt, und wir erweitert diese an Mäuse. Da Mäuse mit den niedrigsten entwickelten Säugetieren unter Labortiere sind, werden ihre Fähigkeiten oft unterschätzt. Daher ist die überraschendste Aspekt der Daten, dass die Trainingserfolg mit fast allen Mäusen erreicht werden...

Die Entwicklung neuer Verfeinerungsstrategien für Labormäuse ist eine anspruchsvolle Aufgabe, die für die Erfüllung des 3R - Prinzips trägt (Replacement, Reduction und Refinement) von 1 Versuchstierkunde. Verbesserungen im Bereich der Verfeinerung kann weiter auf das Wohlbefinden von Millionen von Tieren beitragen, die zu Versuchszwecken verwendet werden. Daher ist eine intensive Forschung auf dem Gebiet benötigt. Dies ist auch eine definierte Ziel der Richtlinie 2010/63 der Europäischen Union. Richtlinie 2010/63 / EU weist darauf hin , dass die Lebenserfahrung von Labortieren verbessert werden muss und dass (Artikel 3.7) 2 &quot;Betriebe wird Gewöhnung und Ausbildungsprogramme geeignet für die Tiere, die Verfahren und die Dauer des Projekts einrichten&quot;. Labortiere können viele Stress-Situationen erleben, während sie gehalten und für Experimente gezüchtet werden. Üblicherweise wird die Wechselwirkung zwischen dem ArbeitsAtory Mäuse und die Verantwortlichen ist eher begrenzt. Daher kann eine vertrauensvolle Beziehung nicht entwickeln. Dies kann erhöhte Angst und Stress in Reaktion auf Behandlung hervorzurufen, die das Verhalten und die Physiologie schädlich ist, und deshalb das Wohlbefinden der Tiere 3, 4, 5, 6. Darüber hinaus routinemäßig durchgeführt Laborverfahren wie allgemeine Handhabung, Zurückhaltung und Blut oder Gewebeentnahme kann Stressreaktionen hervorrufen, die durch die Messung verschiedener Parameter untersucht werden können, wie zum Beispiel Stresshormone oder Verhalten 7, 8. Es hat sich gezeigt , dass die Handhabung Programme effizient 9 die Angst in Richtung der Ermittler in Labornagern verringern kann, 10, 11. Handhabung Programme können deshalb die Tiere zu verbessern &#39;Bedingungen und könnte erheblich zum Tierschutz 5 beitragen. Das Ziel dieser Studie ist es positive Verstärkung Ausbildung für Mäuse als eine spezifische Behandlung Programm einzuführen. Positive Verstärkung Ausbildung ist eine Form der operanten Konditionierung, dass der Prüfer ein Mittel gibt das Verhalten der Tiere zu gestalten. Wenn das Tier ein gewünschtes Verhalten durchführt, wird es durch einen positiven Reiz (hier eine Futterbelohnung) verfolgt. Die Absicht ist, dass das Tier die Belohnung auf das jeweilige Verhalten verbindet. Clicker - Training ist eine Form der positiven Verstärkung Ausbildung eines konditionierten sekundären reinforcer mit der &quot;Klick&quot; -Geräusch eines Klicker und hat sich bewährt 12 ein bestimmtes Verhalten zu stärken. Genauer gesagt, dient der Klick - Sound als &quot;Zeitbrücke&quot; zwischen dem Verhalten und der bevorstehenden Belohnung 13. Der Trainer klickt gerade dann, wenn das Tier führt die gewünschteVerhalten, ohne Zeitverzögerung 14, und dann stellt die Futterbelohnung. Dies stärkt die belohnt Verhalten, das mit einer höheren Frequenz durchgeführt wird. Clickertraining ist weit verbreitet mit Haustieren eingesetzt und hat seinen Weg in die Versuchstierkunde gemacht, wo es 13 mit nicht - menschlichen Primaten erfolgreich umgesetzt wurde, 15, 16. Da Mäuse ziemlich schnell lernen , wenn sie mit der operanten Konditionierung Paradigmen in Frage gestellt, überfordern die Einführung eines zweiten reinforcer sollte nicht ihre kognitiven Fähigkeiten 5, 17, 18, 19. Durch die Einführung von Clicker-Training für die Haltung von Mäusen ermöglichen wir Mäuse kognitive Bereicherung zu erleben. Die Gestaltung der kognitiven Bereicherung müssen die Mäuse ermöglichen, ihre kognitiven Fähigkeiten zu nutzen, um solve Probleme und 20 die Kontrolle über ihre Umgebung zu gewinnen, 21. Mehrere Studien mit verschiedenen Arten belegen die positiven Auswirkungen der kognitiven Anreicherung auf das Wohlergehen der Tiere in Gefangenschaft 22, 23, 24. Die Verbesserung der Fähigkeit der Tiere mit ökologischen Herausforderungen erfolgreich zu meistern trägt zu deren Wohlbefinden 25, 26. Darüber hinaus, wenn die Tiere geringe Mengen an Stress im Laufe ihres Lebens erleben, sind sie weniger anfällig für schädliche Bewältigungsstrategien zu entwickeln, wenn sie mit Stressoren konfrontiert, die in der biomedizinischen Forschung auftreten. Somit kann die konsequente Umsetzung der kognitiven Bereicherung zu einer Homogenisierung der Phänotypen &quot;Themen beitragen. Dies wird zur 3R Prinzip der Reduktion beitragen, da sie die Anzahl der Probanden reduzieren kann erforderlichstatistischen Anforderungen gerecht zu werden 27. Durch ein zuverlässiges Protokoll zu entwickeln, die leicht in den Alltag Labormäuse halten integriert werden können, können wir im Wesentlichen ihre Lebenserfahrung der Verbesserung des Wohlergehens.

<table><tbody><tr><td>Lid for open housing</td><td>Tecniplast</td><td>GM500LID117</td><td></td></tr><tr><td>SealSAfe Plus top for open housing</td><td>Tecniplast</td><td>GM500400SU</td><td></td></tr><tr><td>Type II long, filter top cages</td><td>Tecniplast</td><td>GM500PFSPC</td><td></td></tr><tr><td>Aspen bedding material&amp;nbsp;</td><td>Lab &amp;amp; Vet Service GmbH</td><td>H0234-300</td><td>Environmental enrichment</td></tr><tr><td>Red polycarbonate Mouse House</td><td>Tecniplast</td><td>ACRE011</td><td>Environmental enrichment</td></tr><tr><td>Tissue papers</td><td>Tork, SCA Hygiene Products GmbH</td><td>290179</td><td>Environmental enrichment</td></tr><tr><td>Food - ssniff M-H Extrudat</td><td>ssniff</td><td>V1126-000</td><td>ad libitum</td></tr><tr><td>Target Stick with Clicker</td><td>Trixie</td><td>2282</td><td></td></tr><tr><td>PVC Tube (Tunnels)&amp;nbsp;</td><td>Thyssen Krupp</td><td>RTPVCU04003005</td><td></td></tr><tr><td>White Chocolate/ white chocolate cream</td><td></td><td></td><td>Company doesn&#039;t matter, preferable organic quality</td></tr><tr><td>Forceps</td><td>FineScienceTools</td><td>&lt;em&gt;e.g.&lt;/em&gt; 11150-10</td><td>Or any other tool to fixate chocolate</td></tr><tr><td>Prism Version 6.0 for Windows&amp;nbsp;</td><td>GraphPad Software</td><td></td><td></td></tr></tbody></table>

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.

Die erste und auch einer der wichtigsten Schritte, war die Bestimmung eines geeigneten Futterbelohnung. Daher wurden die Mäuse verschiedene Arten von Nüssen angeboten, einer Zuckerlösung, Marmelade, und verschiedene Arten von Schokolade in einer Petrischale (Abbildung 1). Nach unseren Erfahrungen zeigten die Mäuse eine offensichtliche Vorliebe für weiße Schokolade. Daher haben wir weiße Schokolade für alle weiteren Bildungsprozesse. <p class="jove_content" fo...

Watch this Scientific Journal Video about Introducing Clicker Training as a Cognitive Enrichment for Laboratory Mice at JoVE.com

Introducing Clicker Training as a Cognitive Enrichment for Laboratory Mice

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 ...

introducing-clicker-training-as-cognitive-enrichment-for-laboratory

Universidad San Sebastian

Research

JoVE Journal

Behavior

12.8K Aufrufe.  Johannes Gutenberg-Universität Mainz. 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. 

Serie: Introducing Clicker Training as a Cognitive Enrichment for Laboratory Mice

Automated, Quantitative Cognitive/Behavioral Screening of Mice: For Genetics, Pharmacology, Animal Cognition and Undergraduate Instruction

We describe a high-throughput, high-volume, fully automated, live-in 24/7 behavioral testing system for assessing the effects of genetic and pharmacological manipulations on basic mechanisms of cognition and learning in mice. A standard polypropylene mouse housing tub is connected through an acrylic tube to a standard commercial mouse test box. The test box has 3 hoppers, 2 of which are connected to pellet feeders. All are internally illuminable with an LED and monitored for head entries by infrared (IR) beams. Mice live in the environment, which eliminates handling during screening. They obtain their food during two or more daily feeding periods by performing in operant (instrumental) and Pavlovian (classical) protocols, for which we have written protocol-control software and quasi-real-time data analysis and graphing software. The data analysis and graphing routines are written in a MATLAB-based language created to simplify greatly the analysis of large time-stamped behavioral and physiological event records and to preserve a full data trail from raw data through all intermediate analyses to the published graphs and statistics within a single data structure. The data-analysis code harvests the data several times a day and subjects it to statistical and graphical analyses, which are automatically stored in the &#34;cloud&#34; and on in-lab computers. Thus, the progress of individual mice is visualized and quantified daily. The data-analysis code talks to the protocol-control code, permitting the automated advance from protocol to protocol of individual subjects. The behavioral protocols implemented are matching, autoshaping, timed hopper-switching, risk assessment in timed hopper-switching, impulsivity measurement, and the circadian anticipation of food availability. Open-source protocol-control and data-analysis code makes the addition of new protocols simple. Eight test environments fit in a 48 in x 24 in x 78 in&#160;cabinet; two such cabinets (16 environments) may be controlled by one computer.

Fully automated system for measuring physiologically meaningful properties of the mechanisms mediating spatial localization, temporal localization, duration, rate&#160;and probability estimation, risk assessment, impulsivity, and the accuracy and precision of memory, in order to assess the effects of genetic and pharmacological manipulations on foundational mechanisms of cognition in mice.

Automatisierte, Quantitative Kognition / Verhalten Screening der Mäuse: Für Genetik, Pharmakologie, Animal Cognition und Undergraduate-Unterricht

We describe a high-throughput, high-volume, fully automated, live-in 24/7 behavioral testing system for assessing the effects of genetic and ...

Forschung

Verhalten

A Fully Automated Rodent Conditioning Protocol for Sensorimotor Integration and Cognitive Control Experiments

Rodents have been traditionally used as a standard animal model in laboratory experiments involving a myriad of sensory, cognitive, and motor tasks. Higher cognitive functions that require precise control over sensorimotor responses such as decision-making and attentional modulation, however, are typically assessed in nonhuman primates. Despite the richness of primate behavior that allows multiple variants of these functions to be studied, the rodent model remains an attractive, cost-effective alternative to primate models. Furthermore, the ability to fully automate operant conditioning in rodents adds unique advantages over the labor intensive training of nonhuman primates while studying a broad range of these complex functions.
Here, we introduce a protocol for operantly conditioning rats on performing working memory tasks. During critical epochs of the task, the protocol ensures that the animal's overt movement is minimized by requiring the animal to 'fixate' until a Go cue is delivered, akin to nonhuman primate experimental design. A simple two alternative forced choice task is implemented to demonstrate the performance. We discuss the application of this paradigm to other tasks.

A fully automated protocol for rodent operant conditioning is proposed. The protocol relies on precise temporal control of behavioral events to investigate the extent to which this control influences neural activity underlying sensorimotor integration and cognitive control experiments.

Eine vollautomatische Nageranlage Protokoll für die Integration sensorischer und kognitiver Kontrollexperimente

Rodents have been traditionally used as a standard animal model in laboratory experiments involving a myriad of sensory, cognitive, and motor tasks. ...

The Attentional Set Shifting Task: A Measure of Cognitive Flexibility in Mice

Cognitive impairment, particularly involving dysfunction of circuitry within the prefrontal cortex (PFC), represents a core feature of many neuropsychiatric and neurodevelopmental disorders, including depression, post-traumatic stress disorder, schizophrenia and autism spectrum disorder. Deficits in cognitive function also represent the most difficult symptom domain&#160;to successfully treat, as serotonin reuptake inhibitors and tricyclic antidepressants have only modest effects. Functional neuroimaging studies and postmortem analysis of human brain tissue implicate the PFC as being a primary region of dysregulation in patients with these disorders. However, preclinical behavioral assays used to assess these deficits in mouse models which can be readily manipulated genetically and could provide the basis for studies of new treatment avenues have been underutilized. Here we describe the adaptation of a behavioral assay, the attentional set shifting task (AST), to be performed in mice to assess prefrontal cortex mediated cognitive deficits. The neural circuits underlying behavior during the AST are highly conserved across humans, nonhuman primates and rodents, providing excellent face, construct and predictive validity.

The goal of this protocol is to perform a behavioral assay such as the attentional set shifting task (AST) to assess prefrontal cortex-mediated cognitive flexibility in mice.

Die Aufmerksamkeits Set Shifting Aufgabe: Eine Messung der kognitiven Flexibilität bei Mäusen

Cognitive impairment, particularly involving dysfunction of circuitry within the prefrontal cortex (PFC), represents a core feature of many ...

Operant Sensation Seeking in the Mouse

Operant methods are powerful behavioral tools for the study of motivated behavior. These 'self-administration' methods have been used extensively in drug addiction research due to their high construct validity. Operant studies provide researchers a tool for preclinical investigation of several aspects of the addiction process. For example, mechanisms of acute reinforcement (both drug and non-drug) can be tested using pharmacological or genetic tools to determine the ability of a molecular target to influence self-administration behavior1-6. Additionally, drug or food seeking behaviors can be studied in the absence of the primary reinforcer, and the ability of pharmacological compounds to disrupt this process is a preclinical model for discovery of molecular targets and compounds that may be useful for the treatment of addiction3,7-9. One problem with performing intravenous drug self-administration studies in the mouse is the technical difficulty of maintaining catheter patency. Attrition rates in these experiments are high and can reach 40% or higher10-15. Another general problem with drug self-administration is discerning which pharmacologically-induced effects of the reinforcer produce specific behaviors. For example, measurement of the reinforcing and neurological effects of psychostimulants can be confounded by their psychomotor effects. Operant methods using food reinforcement can avoid these pitfalls, although their utility in studying drug addiction is limited by the fact that some manipulations that alter drug self-administration have a minimal impact on food self-administration. For example, mesolimbic dopamine lesion or knockout of the D1 dopamine receptor reduce cocaine self-administration without having a significant impact on food self-administration 12,16. 
Sensory stimuli have been described for their ability to support operant responding as primary reinforcers (i.e. not conditioned reinforcers)17-22. Auditory and visual stimuli are self-administered by several species18,21,23, although surprisingly little is known about the neural mechanisms underlying this reinforcement. The operant sensation seeking (OSS) model is a robust model for obtaining sensory self-administration in the mouse, allowing the study of neural mechanisms important in sensory reinforcement24. An additional advantage of OSS is the ability to screen mutant mice for differences in operant behavior that may be relevant to addiction. We have reported that dopamine D1 receptor knockout mice, previously shown to be deficient in psychostimulant self-administration, also fail to acquire OSS24. This is a unique finding in that these mice are capable of learning an operant task when food is used as a reinforcer. While operant studies using food reinforcement can be useful in the study of general motivated behavior and the mechanisms underlying food reinforcement, as mentioned above, these studies are limited in their application to studying molecular mechanisms of drug addiction. Thus, there may be similar neural substrates mediating sensory and psychostimulant reinforcement that are distinct from food reinforcement, which would make OSS a particularly attractive model for the study of drug addiction processes. The degree of overlap between other molecular targets of OSS and drug reinforcers is unclear, but is a topic that we are currently pursuing. While some aspects of addiction such as resistance to extinction may be observed with OSS, we have found that escalation 25 is not observed in this model24. Interestingly, escalation of intake and some other aspects of addiction are observed with self-administration of sucrose26. Thus, when non-drug operant procedures are desired to study addiction-related processes, food or sensory reinforcers can be chosen to best fit the particular question being asked. 
In conclusion, both food self-administration and OSS in the mouse have the advantage of not requiring an intravenous catheter, which allows a higher throughput means to study the effects of pharmacological or genetic manipulation of neural targets involved in motivation. While operant testing using food as a reinforcer is particularly useful in the study of the regulation of food intake, OSS is particularly apt for studying reinforcement mechanisms of sensory stimuli and may have broad applicability to novelty seeking and addiction.

In this protocol we describe a method of operant learning using sensory stimuli as a reinforcer in the mouse. It requires no prior training or food restriction, and it allows the study of motivated behavior without the use of a pharmacological or natural reinforcer such as food.

Operante Sensation Seeking in der Maus

Operant methods are powerful behavioral tools for the study of motivated behavior.  These 'self-administration' methods have been used extensively in drug ...

Neurowissenschaften

Quantifying Social Motivation in Mice Using Operant Conditioning

In this protocol, social motivation is measured in mice through a pair of operant conditioning paradigms. To conduct the experiments, two-chambered shuttle boxes were equipped with two operant levers (left and right) and a food receptacle in one chamber, which was then divided from the second chamber by an automated guillotine door covered by a wire grid. Different stimulus mice, rotated across testing days, served as a social stimulus behind the wire grid, and were only visible following the opening of the guillotine door. Test mice were trained to lever press in order to open the door and gain access to the stimulus partner for 15 sec. The number of lever presses required to obtain the social reward progressively increased on a fixed schedule of 3. Testing sessions ended after test mice stopped lever pressing for 5 consecutive minutes. The last reinforced ratio or breakpoint can be used as a quantitative measure of social motivation. For the second paradigm, test mice were trained to discriminate between left and right lever presses in order to obtain either a food reward or the social reward. Mice were rewarded for every 3 presses of each respective lever. The number of food and social rewards can be compared as a measurement of the value placed upon each reward. The ratio of each reward type can also be compared between mouse strains and the change in this ratio can be monitored within testing sessions to measure satiation with a given reward type. Both of these operant conditioning paradigms are highly useful for the quantification of social motivation in mouse models of autism and other disorders of social behavior.

This article describes a new protocol for the quantitative measurement of social motivation in mice using operant conditioning for a social reward. The protocol is useful for the measurement of social motivation in mouse models of autism and other disorders of social behavior.

Quantifizierung Social Motivation in Mäuse mit Operante Konditionierung

In this protocol, social motivation is measured in mice through a pair of operant conditioning paradigms. To conduct the experiments, two-chambered ...

A Fully Automated and Highly Versatile System for Testing Multi-cognitive Functions and Recording Neuronal Activities in Rodents

We have developed a fully automated system for operant behavior testing and neuronal activity recording by which multiple cognitive brain functions can be investigated in a single task sequence. The unique feature of this system is a custom-made, acoustically transparent chamber that eliminates many of the issues associated with auditory cue control in most commercially available chambers. The ease with which operant devices can be added or replaced makes this system quite versatile, allowing for the implementation of a variety of auditory, visual, and olfactory behavioral tasks. Automation of the system allows fine temporal (10 ms) control and precise time-stamping of each event in a predesigned behavioral sequence. When combined with a multi-channel electrophysiology recording system, multiple cognitive brain functions, such as motivation, attention, decision-making, patience, and rewards, can be examined sequentially or independently.

In this report, we present a fully automated and highly versatile system capable of simultaneously testing multi-cognitive behaviors and recording neuronal activities for rodents.

Ein vollautomatisiertes und vielseitig verwendbares System zum Testen von Multi-kognitiven Funktionen und Aufnahme neuronaler Aktivitäten bei Nagetieren

We have developed a fully automated system for operant behavior testing and neuronal activity recording by which multiple cognitive brain functions can be ...

Using Clicker Training and Social Observation to Teach Rats to Voluntarily Change Cages

Cage cleaning is a routinely performed husbandry procedure and is known to induce stress in laboratory rats. As stress can have a negative impact on well-being and can affect the comparability and reproducibility of research results, the amount of stress experienced by laboratory animals should be minimized and avoided when possible. Further, the direct contact between the rat and animal caretaker during the cage change bears hygiene risks and therefore possibly negatively impacts the well-being of the rats and the quality of the research. 
Our protocol aims to improve the routinely performed cage changing procedure. For this reason, we present a feasible protocol that enables rats to learn via clicker training and observation to voluntarily change to a clean cage. This training helps to reduce stress caused by the physical disturbance and handling associated with the cage changes and concurrently enables a reduction in direct contact between animal and animal caretaker after the training phase is completed.
The implementation of clicker training to rats is fast and easy. Rats are generally interested in the training and efficiently learn the desired behavior, which entails changing cages through a pipe. Even without training, the rats learn to perform the desired behavior by observation, as 80% of the observational learning group successfully changed cages when tested. The training further helps to establish a relationship of trust between trainer and animal. As hygiene and well-being are both very important in animal experiments, this protocol might also help to improve high-quality research.

This protocol introduces a method of cage change for rats via clicker training. Rats learn the desired behavior not only by direct training but also by observational learning. The implementation of this fast and easy protocol might help to improve well-being and hygiene in rodent facilities.

Verwendung von Clicker-Training und soziale Beobachtung beibringen Ratten Käfige freiwillig ändern

Cage cleaning is a routinely performed husbandry procedure and is known to induce stress in laboratory rats. As stress can have a negative impact on ...

Evaluating Skilled Prehension in Mice Using an Auto-Trainer

We describe a method to introduce na&#239;ve mice to a novel prehension (reach-to-grasp) task. Mice are housed singly in cages with a frontal slot that permits the mouse to reach out of its cage and retrieve food pellets. Minimal food restriction is employed to encourage the mice to perform the food retrieval from the slot. As the mice begin to associate coming to the slot for food, the pellets are manually pulled away to stimulate extension and pronation of their paw to grasp and retrieve the pellet through the frontal slot. When the mice begin to reach for the pellets as they arrive at the slot, the behavioral assay can be performed by measuring the rate at which they successfully grasp and retrieve the desired pellet. They are then introduced to an auto-trainer that automates both the process of providing food pellets for the mouse to grasp, and the recording of successful and failed reaching and grasping attempts. This allows for the collection of reaching data for multiple mice with minimal effort, to be used in experimental analysis as appropriate.

Method to assess the impact of training on motor skills is a useful tool. Unfortunately, most behavioral assessments can be labor intensive and/or expensive.We describe here a robotic method of assessing prehension (reach-to-grasp) skill in mice.

Bewertung der geschickten Vorhension bei Mäusen mit einem Auto-Trainer

We describe a method to introduce na&#239;ve mice to a novel prehension (reach-to-grasp) task. Mice are housed singly in cages with a frontal slot that ...

Positives Verstärkungstraining für routinemäßige Kaninchenhaltungsverfahren

Training Laboratory Rabbits to Refine Routine Husbandry Procedures

Non-aversive handling and training techniques for laboratory animals are required to facilitate experimental and routine husbandry procedures, improving both animal welfare and scientific quality. Clicker training was utilized to develop training protocols for rabbits to refine stressful routine husbandry procedures usually associated with lifting (i.e., being picked up from the floor)/restraining (i.e., being held in the arms of a human) them. Thirteen female New Zealand White rabbits were trained over three weeks. All rabbits learned the predefined goal behaviors: they followed the target stick, jumped onto the weighing scale, entered a transport box, and reared while placing their front paws onto the trainer&#39;s hand. In addition, ten animals jumped from the floor onto the sitting trainer&#39;s lap and allowed the trainer to lift their paws off the surface while sitting on the trainer&#39;s lap. For some individuals, the protocols had to be adapted by additional interim steps. At the end of the training, the rabbits reliably showed the expected goal behaviors, even after short and long training breaks. With few exceptions, a familiar person other than the trainer could elicit the goal behaviors from the rabbits (generalization), though further sessions were required for generalization. In the voluntary approach test, the rabbits preferred interacting with the trainer in the 1st trial but spent as much time with an unfamiliar person as with the trainer in the 2nd trial. The behavioral observations suggested that picking the rabbits up with the transport box, as described in the protocol, instead of restraining them with the scruff of their neck and lifting them on the arm, was less aversive. All in all, the training protocols were feasible and can serve as a refinement strategy in laboratory animal facilities. In the interest of animal welfare, the training protocols should be applied wherever possible.

Autor im Rampenlicht: Training von Versuchstieren für einen schonenden und stressfreien Umgang

Training protocols involving positive reinforcement for routine husbandry procedures in rabbits were developed and proved to be successful, as the rabbits reliably showed the trained behaviors at the weekly general examination. Picking the rabbits up with the transport box was less aversive than the conventional handling technique.

Training von Laborkaninchen zur Verfeinerung routinemäßiger Haltungsverfahren

Non-aversive handling and training techniques for laboratory animals are required to facilitate experimental and routine husbandry procedures, improving ...

Einführung in Clicker Ausbildung als Cognitive Anreicherung für Labormäuse

Zusammenfassung

Weitere Videos entdecken

Kapitel in diesem Video

Automatisierte, Quantitative Kognition / Verhalten Screening der Mäuse: Für Genetik, Pharmakologie, Animal Cognition und Undergraduate-Unterricht

Eine vollautomatische Nageranlage Protokoll für die Integration sensorischer und kognitiver Kontrollexperimente

Die Aufmerksamkeits Set Shifting Aufgabe: Eine Messung der kognitiven Flexibilität bei Mäusen

Operante Sensation Seeking in der Maus

Quantifizierung Social Motivation in Mäuse mit Operante Konditionierung

Ein vollautomatisiertes und vielseitig verwendbares System zum Testen von Multi-kognitiven Funktionen und Aufnahme neuronaler Aktivitäten bei Nagetieren

Verwendung von Clicker-Training und soziale Beobachtung beibringen Ratten Käfige freiwillig ändern

Bewertung der geschickten Vorhension bei Mäusen mit einem Auto-Trainer

Training von Laborkaninchen zur Verfeinerung routinemäßiger Haltungsverfahren

Training von Laborkaninchen zur Verfeinerung routinemäßiger Haltungsverfahren