Diese Arbeit wird durch einen Zuschuss der Baylor University Research Council und von einem Forschungsstipendium von der Epilepsie-Stiftung unterstützt.

Die in dem folgenden Versuch verwendeten Mäuse wurden erzeugt und an der Baylor University untergebracht bei einer Raumtemperatur von 22 ° C, mit einem 14 Stunden Licht und 10 Stunden Dunkelheit (von 20.00 bis 06.00 h) Tagesgang. Die Mäuse wurden ad libitum Zugang zu Futter und Wasser. Alle Eingriffe, die die Mäuse waren in Übereinstimmung mit den National Institutes of Health Leitlinien für die Pflege und Verwendung von Labortieren und das Tier-Protokoll wurde von der Baylor University Animal Care und Verwenden Committee genehmigt. Überblick Die Spur Anlage Angst Aufgabe wird durch Wiltgen und Kollegen 9 beschriebenen Verfahren. 1. Vorbereitung der Ausrüstung Die Angst Anlagengerät Kammer (26 cm x 22 cm x 18 cm) besteht aus zwei Seiten, die Acryl sind, zwei Seiten, die Metall sind, und ein Gitterboden Boden, der verwendet wird, um eine leichte Fuß-Schock zu liefern. Die Testkammer ist Hausd in einer gedämpften Klangkammer. Die Kammer ist auch lichtdicht nach außen Licht von Beeinflussung der Bewegungserkennung-Software zu verhindern. <ol><li> Kalibrieren Sie die Schlagniveaus, Lichtverhältnisse und Schallintensitätspegel für die Testkammer. Messen Sie die Hintergrundebene der Testkammer. Das Hintergrundrauschen in dieser Kammer ist 65 dB. Verwenden Sie eine Lärmmessung, um dieses Niveau zu messen. Das Messgerät sollte auf 70 dB eingestellt werden, stellen nach C, und die Aufdeckung Einstellung langsam. <ol><li> Kalibrieren Sie den Schock-Ebene auf 0,5 mA. Verwenden Sie eine externe Kalibrierungseinrichtung, um den Schock Ebene (siehe Tabelle Materialien) richtig zu kalibrieren. Die interne Messung der Schockgenerator ist nicht korrekt. Der Schock-Generator verwaltet einen verschlüsselten Schock, der nicht genau durch eine Standard-Amperemeter gemessen werden kann. </li><li> Legen Sie eine Führung auf einem der Gitterstäbe und legen Sie die andere Leitung 3 oder 4 Bars vorbei. Verwenden Sie den Schock-Generator, um den Schock zu verabreichen. Stellen Sie die Schockstufe, bis die richtige Höhe erreicht ist. Tun Sie dies für each fürchten Klimakammer. </li><li> Schließen Sie die Tür zu der Kammer bei der Messung der Lichtstärke der Kammer. Kalibrieren Sie den Lichtpegel auf ca. 1,0. Dies ist eine Zahl, die spezifisch für die Freeze Software in diesem Experiment verwendet. Ein externer Belichtungsmesser wird dies als 2 Lux lesen. Die Lichtstärke kann durch Verschieben der Lage des Hauses Licht oder durch Einstellen des Kondensators der Linse eingestellt werden. Stellen Sie sicher, dass die Stellschrauben für die Linse nach der Einstellung anziehen. </li><li> Kalibrieren Sie den Klang Dezibel-Pegel um 85 dB. Verwenden Sie eine externe Dezibel-Messer in der Testkäfig um den Dezibel-Pegel (siehe Materialien Table) zu kalibrieren. Die vorgestellte Sound wird ein 2700-Hz-Ton sein. Hinweis: Wenn Sie Mäuse älter als 6 Monate alt ist, kann es besser sein, weißes Rauschen zu verwenden, da ältere Mäuse können Gehördefizite. </li></ol></li><li> Nachdem das Gerät bereit ist, nehmen die Themen auf eine separate Halteraum. Hinweis: Lassen Sie die Maus Haus nicht im selben Raum wie der Testraum. </li><li> Label-the Schwanz der Mäuse, die in der Reihenfolge der Tests getestet werden. Es ist am besten, um übermäßige Handhabung vor der Prüfung zu reduzieren. Alternativ können die Schwänze beschriftet am Tag vor dem Experiment zu reduzieren Umgang mit Stress werden. Nachdem die Mäuse wurden markiert damit sie in den Raum 30 Minuten akklimatisieren. Haben extra sauber Käfige für die Unterbringung der Mäuse nach der Prüfung ist abgeschlossen. </li></ol> 2. Trace Anlage Tag 1 <ol><li> Entfernen Sie jede Maus aus dem Käfig und sie in Einzelkäfige für den Transport zur Angst Konditionierungskammer. Saubere Bettwäsche Verwenden Sie für jeden Käfig. Legen Sie eine Notiz auf dem Transfer Käfig, um die Reihenfolge der Mäuse richtig testen zu halten. Hinweis: Wenn die Mäuse werden einzeln untergebracht, dann können sie in ihrem eigenen Käfig transportiert werden. </li><li> Platzieren Sie die Maus in der Testkammer und die Tür schließen. Starten Sie das Software-Programm. </li><li> Am Trainingstag, damit die Mäuse, um die Kammer für 3 min zu erkunden. Die Software stellt dann einen 20 Sekunden Ton (85 dB, 2700 Hz) to das Tier. Nach einer Periode der Spur 20 sec ein mildes Schock (2 sec, 0,5 mA) an das Tier verabreicht. </li><li> Notieren Sie die Reaktion des Subjekts zu bestätigen, dass sie erhalten die aversiven Reizen, indem Sie das Video. A 200 sec inter-Testintervall trennt die 5 Anlage Studien. Jeder Versuch besteht aus einem 20 Sekunden Ton gefolgt von einem 20 Sekunden Verzögerung dann ein Schock. </li><li> Nachdem der Test abgeschlossen ist, um damit die Tiere in der Testkammer für 1 Minute, bevor es von der Testkäfig entfernt bleiben. </li><li> Legen Sie das Tier wieder in den Käfig und Transfer sie wieder in ihre Heimat Käfig. Wenn es zusätzliche Mäuse in ihrem Heimkäfig dann einzeln mit der Maus aufnehmen, bis alle Mäuse vollständige Prüfung. Dadurch wird die Belastung auf die andere Mäuse, die nicht getestet wurden, zu reduzieren. Eine alternative Lösung wäre, alle Mäuse einzeln Haus eine Woche vor der Prüfung um den Einfluss der kontinuierlichen Entfernung von Mäusen aus einem Käfig zu reduzieren. </li><li> Reinigen Sie die Testkammer mit 30% Isopropanol nach each Tier getestet. </li><li> Wiederholen Sie die Schritte 2,2-2,7 für alle Mäuse in der Test Kohorte. </li><li> Gibt alle Mäuse, um ihre Kolonie Raum nach der letzten Maus in der Kohorte getestet. </li></ol> 3. Trace Angstkonditionierung Tag 2: Trace Memory Testing <ol><li> Trace-Speicher Tests werden an Tag 2 auf. In diesem Protokoll gibt es 3-Ton-Präsentationen. Legen Sie die Mäuse in einen neuen Kontext für die Trace-Testanlage. </li><li> Bereiten Sie die Software, ein Programm mit einer 2 min Basisperiode, gefolgt von drei 20 Sekunden Ton Präsentationen laufen. Es gibt einen 220 sec ITI zwischen jedem Ton Präsentation. </li><li> Für den neuen Kontext Zustand, legen klarem Acryl-Einsätze auf dem Boden der Kammer, um die Form, Textur und Farbe der Konditionierungskammer verändern. </li><li> Ändern Sie den Geruch in der Kammer, indem Vanille-Extrakt in einem Boot wiegen unter dem Boden-Einsatz. </li><li> Reinigen der Kammer mit 70% Ethanol anstelle von 30% igem Isopropanol. Hinweis: Dies wird helfen,die Bereitstellung eines neuen Kontext zu erstellen. </li><li> Bringen Sie die Mäuse mit dem Halteraum und umbenennen ihre Schwänze zum Testen, wenn nötig. </li><li> Bereiten Roman Kontextübertragung Käfige durch das Bettzeug mit Papierschnitzel ersetzt. Hinweis: Dies wird bei der Schaffung eines neuartigen Kontext zu helfen. </li><li> Legen Sie die Mäuse in der Testkammer dann das Programm starten. Reinigen der Kammer mit 70% Ethanol, nachdem der Test abgeschlossen ist. </li><li> Bringen Sie die Mäuse in ihre Heimat Käfig, nachdem die Spurenanlage ist abgeschlossen. Gibt alle Mäuse, um ihre Kolonie Raum, wenn alle Mäuse getestet wurden. </li></ol> 4. Trace Angstkonditionierung Tag 3: Kontext Memory Testing <ol><li> Am dritten Tag kontextuellen Anlage durchgeführt wird. Bereiten Sie die Software, ein Programm zum Gefrierverhalten für 8 Minuten aufzeichnen laufen. </li><li> Reinigen Sie die Kammer mit 30% Isopropanol vor der Prüfung und nach der Prüfung jeder Maus. Der Zusammenhang ist identisch mit der von Tag 1 sein. Die Übertragungs Käfige sollten die gleichen wie in 1 Tag sein. </li><li&gt; Bringen Mäuse an der Halteraum und beschriften ihre Schwänze zum Testen. </li><li> Platzieren Sie jede Maus in die Testkammer dann das Programm starten. Reinigen Sie die Kammer mit 30% Isopropanol nach der Prüfung ist abgeschlossen. </li><li> Gibt alle Mäuse, um ihre Kolonie Zimmer, als abgeschlossen. </li></ol>

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Die Autoren haben nichts zu offenbaren.

Es gibt mehrere Studien, die die neuronalen Schaltkreise, die Spur Angst zugrunde liegt Anlage aufgeklärt haben. Trace Furchtkonditionierung wird angenommen, dass die CA1 des Hippocampus 12-14 beinhalten. Es gibt auch Hinweise, dass die medialen präfrontalen Kortex (mPFC) spielt eine große Rolle in der Spurenaugenblinkanlage 15 und die mPFC wurde festgestellt, in der Spuren Angst Anlage einbezogen werden. Eine Studie ergab, dass mPFC Neuronen bieten anhaltende Aktivität während der Ablaufverfo...

In Angst Anlage ein neutraler Reiz (NS) mit einem aversiven unkonditionierten Stimulus (US) gepaart. Die NS ist normalerweise ein Ton und wird zu einem bedingten Reiz (CS) durch wiederholte Paarungen mit den USA. Der CS kann dann rufen eine konditionierte Reaktion (CR), wie Einfrieren, in Abwesenheit der aversiven USA. Eine häufig verwendete Protokoll ist Angst Anlage Verzögerung Anlage. In diesem Protokoll der Beginn der NS und der US angrenzt oder mit etwas Überlappung Reizdarbietung. Auch wenn Verzögerung Angst Anlage ist eine der am häufigsten verwendeten Arten von zeitlichen assoziativen Anlage, gibt es mehrere andere Arten von assoziativen Anlage zeitliche Arrangements: Simultananlage, rückwärts Anlage und Spurenanlage ein. In Spur Angst Anlage gibt es einen Reiz freien Intervall zwischen dem NS und den USA von mehreren Sekunden, was zu einer &quot;Spur&quot; Zeitraum. Mehrere Studien haben berichtet Defizits in der Spurenangstkonditionierung bei neurotoxische Läsionen in den Strukturen hergestellt, dass die Eingabe in die Hippocampus 2-5 oder wenn pharmakologische Mittel werden verwendet, um die Rezeptorfunktion im Hippocampus zu blockieren. Läsion Hippocampus führt zu Defiziten in der Spurenanlage und kontextuellen Anlage, aber keine Angst Verzögerungsanlage 8 beeinträchtigen. Es gibt mehrere Vorteile für die Verwendung Spuren Angst Anlage. Die Angst Anlage Protokoll kann über einen Drei-Tage-Testzeitraum erreicht werden und ermöglicht die Hippocampus-abhängigen Speicher, der nicht ortsabhängig. Trace Furchtkonditionierung kann als ergänzende Prüfung mit den Morris-Wasserlabyrinth, neuartige Objekterkennungstest oder andere Labyrinth-Tests bei der Untersuchung von Hippocampus-abhängigen Speicher verwendet werden.

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trace fear conditioning in mice

In diesem Experiment zeigen wir eine Technik zum Lernen und Gedächtnis zu messen. In der hier vorgestellten Spur Angst Anlage Protokoll gibt es fünf Paarungen zwischen einem neutralen Reiz und einem unbedingten Reiz. Es gibt eine 20 Sekunden Zeit, die jede Spur Anlage Studie trennt. Am folgenden Tag Einfrieren während der Präsentation des konditionierten Stimulus (CS) und Spuren Zeitraum gemessen. Am dritten Tag gibt es ein 8-Minuten-Test, um kontextbezogene Speicher messen. Die repräsentativen Ergebnisse von Mäusen, die mit dem aversiven unbedingten Reiz (Schock) präsentiert wurden im Vergleich zu Mäusen, die die Ton-Präsentationen ohne den unbedingten Reiz erhalten. Trace Furchtkonditionierung wurde erfolgreich eingesetzt, um subtile Lern-und Gedächtnisdefizite und Verbesserungen in Mäusen, die nicht mit anderen Angstkonditionierung Methoden gefunden werden, zu erkennen. Diese Art von Angst Anlage wird angenommen, abhängig von Verbindungen zwischen dem medialen präfrontalen Kortex und Hippocampus zu sein. Eine aktuelle Kontroverse ist, ob diese Methode wird angenommen, dass Amygdala-unabhängig sein. Daher ist anderen Angst Testanlage benötigt, um Amygdala-abhängigen Lern-und Gedächtniseffekte, wie durch die Verzögerung Angst Anlage zu prüfen.

Für die repräsentative Ergebnisse präsentieren wir Daten von Steuer C57BL/6J erwachsenen Mäusen, die den neutralen Reiz-Paarungen mit dem unbedingten Reiz (Schock-Bedingung) im Vergleich zu Mäusen, die den neutralen Reiz erhielt aber nicht den unbedingten Reiz (keine Schockzustand) erhalten erhalten. Es ist wichtig, diesen Zustand zu laufen, wenn erste Einrichtung dieser Verhaltenstest, um festzustellen, ob das Protokoll korrekt durchgeführt wurde. Die Daten in Fig. 1 ...

Watch this Scientific Journal Video about Trace Fear Conditioning in Mice at JoVE.com

Trace Fear Conditioning in Mice

Im folgenden Experiment beschreiben wir ein Protokoll für die Spurenangstkonditionierung bei Mäusen. Diese Art des assoziativen Speicher eine Spur, die die Zeit und die neutralen Reiz unbedingten Reiz trennt.

Trace Angstkonditionierung bei Mäusen

In this experiment we present a technique to measure learning and memory. In the trace fear conditioning protocol presented here there are five pairings ...

Research

JoVE Journal

Behavior

32.6K Aufrufe.  Baylor University.  Im folgenden Experiment beschreiben wir ein Protokoll für die Spurenangstkonditionierung bei Mäusen. Diese Art des assoziativen Speicher eine Spur, die die Zeit und die neutralen Reiz unbedingten Reiz trennt. 

Serie: Trace Fear Conditioning in Mice

Contextual and Cued Fear Conditioning Test Using a Video Analyzing System in Mice

The contextual and cued fear conditioning test is one of the behavioral tests that assesses the ability of mice to learn and remember an association between environmental cues and aversive experiences. In this test, mice are placed into a conditioning chamber and are given parings of a conditioned stimulus (an auditory cue) and an aversive unconditioned stimulus (an electric footshock). After a delay time, the mice are exposed to the same conditioning chamber and a differently shaped chamber with presentation of the auditory cue. Freezing behavior during the test is measured as an index of fear memory. To analyze the behavior automatically, we have developed a video analyzing system using the ImageFZ application software program, which is available as a free download at http://www.mouse-phenotype.org/. Here, to show the details of our protocol, we demonstrate our procedure for the contextual and cued fear conditioning test in C57BL/6J mice using the ImageFZ system. In addition, we validated our protocol and the video analyzing system performance by comparing freezing time measured by the ImageFZ system or a photobeam-based computer measurement system with that scored by a human observer. As shown in our representative results, the data obtained by ImageFZ were similar to those analyzed by a human observer, indicating that the behavioral analysis using the ImageFZ system is highly reliable. The present movie article provides detailed information regarding the test procedures and will promote understanding of the experimental situation.

This article presents a protocol for a contextual and cued fear conditioning test using a video analyzing system to assess fear learning and memory in mice.

Kontextuelle und Cued Angst Anlage Test mit einer Video-Analyse-System bei Mäusen

The contextual and cued fear conditioning test is one of the behavioral tests that assesses the ability of mice to learn and remember an association ...

Forschung

Verhalten

Disrupting Reconsolidation of Fear Memory in Humans by a Noradrenergic &#946;-Blocker

The basic design used in our human fear-conditioning studies on disrupting reconsolidation includes testing over different phases across three consecutive days. On day 1 - the fear acquisition phase, healthy participants are exposed to a series of picture presentations. One picture stimulus (CS1+) is repeatedly paired with an aversive electric stimulus (US), resulting in the acquisition of a fear association, whereas another picture stimulus (CS2-) is never followed by an US. On day 2 - the memory reactivation phase, the participants are re-exposed to the conditioned stimulus without the US (CS1-), which typically triggers a conditioned fear response. After the memory reactivation we administer an oral dose of 40 mg of propranolol HCl, a &#946;-adrenergic receptor antagonist that indirectly targets the protein synthesis required for reconsolidation by inhibiting the noradrenaline-stimulated CREB phosphorylation. On day 3 - the test phase, the participants are again exposed to the unreinforced conditioned stimuli (CS1- and CS2-) in order to measure the fear-reducing effect of the manipulation. This retention test is followed by an extinction procedure and the presentation of situational triggers to test for the return of fear. Potentiation of the eye blink startle reflex is measured as an index for conditioned fear responding. Declarative knowledge of the fear association is measured through online US expectancy ratings during each CS presentation. In contrast to extinction learning, disrupting reconsolidation targets the original fear memory thereby preventing the return of fear. Although the clinical applications are still in their infancy, disrupting reconsolidation of fear memory seems to be a promising new technique with the prospect to persistently dampen the expression of fear memory in patients suffering from anxiety disorders and other psychiatric disorders.

Disrupting Reconsolidation of Fear Memory in Humans by a Noradrenergic &amp;#946;-Blocker

Disrupting reconsolidation is a promising approach to dampen the behavioral expression of fear memory in patients with anxiety disorders or posttraumatic stress disorder. In a series of human fear conditioning studies we showed that disrupting reconsolidation by the noradrenergic &#946;-blocker propranolol is very effective in erasing conditioned fear responding.

Die Unterbrechung Rückverfestigung der Angst Gedächtnis beim Menschen durch eine Noradrenerge β-Blocker

The basic design used in our human fear-conditioning studies on disrupting reconsolidation includes testing over different phases across three consecutive ...

Protocol for Studying Extinction of Conditioned Fear in Naturally Cycling Female Rats

Extinction of conditioned fear has been extensively studied in male rodents. Recently, there have been an increasing number of studies indicating that neural mechanisms for certain behavioral tasks and response behaviors are different in females and males. Using females in research studies can represent a challenge because of the variation of gonadal hormones during their estrous cycle. This protocol describes well-established procedures that are useful in investigating the role of estrogen in fear extinction memory consolidation in female rats. Phase of the estrous cycle and exogenous estrogen administration prior to extinction training can influence extinction recall 24 hr later. The vaginal swabbing technique for estrous phase identification described here aids the examination and manipulation of naturally cycling gonadal hormones. The use of this basic rodent model may further delineate the mechanisms by which estrogen can modulate fear extinction memory in females.

Gonadal hormones such as estrogen modulate memory formation in a number of experimental paradigms including fear extinction memory. This protocol describes a set of methods for investigating the influence of gonadal hormones specifically during extinction in naturally cycling females, including estrous cycle monitoring and exogenous estrogen administration.

Protokoll zur Untersuchung Extinction konditionierter Furcht in natürlich Radfahren weiblichen Ratten

Extinction of conditioned fear has been extensively studied in male rodents. Recently, there have been an increasing number of studies indicating that ...

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 ...

Whisker-signaled Eyeblink Classical Conditioning in Head-fixed Mice

Eyeblink conditioning is a common paradigm for investigating the neural mechanisms underlying learning and memory. To better utilize the extensive repertoire of scientific techniques available to study learning and memory at the cellular level, it is ideal to have a stable cranial platform. Because mice do not readily tolerate restraint, they are usually trained while moving about freely in a chamber. Conditioned stimulus (CS) and unconditioned stimulus (US) information are delivered and eyeblink responses recorded via a tether connected to the mouse's head. In the head-fixed apparatus presented here, mice are allowed to run as they desire while their heads are secured to facilitate experimentation. Reliable conditioning of the eyeblink response is obtained with this training apparatus, which allows for the delivery of whisker stimulation as the CS, a periorbital electrical shock as the US, and analysis of electromyographic (EMG) activity from the eyelid to detect blink responses.

The preparation presented here for whisker-signaled eyeblink conditioning in head-fixed mice precisely stimulates specific whiskers while allowing mice to ambulate on a cylindrical treadmill. A whisker stimulation conditioned stimulus (CS) paired with a periorbital shock unconditioned stimulus (US) results in reliable associative learning on this apparatus.

Whisker-signalisierte Eyeblink Klassische Konditionierung in Head-Fest Mäuse

Eyeblink conditioning is a common paradigm for investigating the neural mechanisms underlying learning and memory. To better utilize the extensive ...

Reducing State Anxiety Using Working Memory Maintenance

The purpose of this protocol is to explain how to examine the relationship between working memory processes and anxiety by combining the Sternberg Working Memory (WM) and the threat of shock paradigms. In the Sternberg WM paradigm, subjects are required to maintain a series of letters in the WM for a brief interval and respond by identifying whether the position of a given letter in the series matches a numerical prompt. In the threat of shock paradigm, subjects are exposed to alternating blocks where they are either at risk of receiving unpredictable presentations of a mild electric shock or are safe from the shock. Anxiety is probed throughout the safe and threat blocks using the acoustic startle reflex, which is potentiated under threat (Anxiety-Potentiated Startle (APS)). By conducting the Sternberg WM paradigm during the threat of shock and probing the startle response during either the WM maintenance interval or the intertrial interval, it is possible to determine the effect of WM maintenance on APS.

This protocol demonstrates how to measure anxiety-potentiated startle during the Sternberg Working Memory paradigm.

Verringerung der staatlichen Angst mit Arbeitsspeicher Wartung

The purpose of this protocol is to explain how to examine the relationship between working memory processes and anxiety by combining the Sternberg Working ...

Classical Short-Delay Eyeblink Conditioning in One-Year-Old Children

Classical eyeblink conditioning (EBC) refers to the learned association between a conditioned stimulus (an auditory tone) and an unconditioned stimulus (a puff of air to the cornea). Eyeblink conditioning is often used experimentally to detect abnormalities in cerebellar-dependent learning and memory that underlies this type of associative learning. While experiments in adults and older children are relatively simple to administer using commercial equipment, eyeblink conditioning in infants is more challenging due to their poor compliance, which makes correct positioning of the equipment difficult. To achieve conditioning in one-year-old infants, a custom-made or an adapted commercial system can be used to deliver the air puff to the infant's cornea. The main challenge lies in successfully detecting and classifying the behavioral responses. We report that automated blink detection methods are unreliable in this population, and that conditioning experiments should be analyzed using frame-by-frame analysis of supplementary video camera recordings. This method can be applied to study developmental changes in eyeblink conditioning and to examine whether this paradigm can detect children with neurological disorders.

This protocol describes an eyeblink conditioning paradigm appropriate for experiments with one-year-old infants. Commercial or custom-made equipment can be used to deliver the stimuli, and data collection and analysis should be performed on the video recordings.

Klassische kurze Verzögerung Eyeblink Konditionierung in One-jährige Kinder

Classical eyeblink conditioning (EBC) refers to the learned association between a conditioned stimulus (an auditory tone) and an unconditioned stimulus (a ...

Stress-Enhanced Fear Learning, a Robust Rodent Model of Post-Traumatic Stress Disorder

Fear behaviors are important for survival, but disproportionately high levels of fear can increase the vulnerability for developing psychiatric disorders such as post-traumatic stress disorder (PTSD). To understand the biological mechanisms of fear dysregulation in PTSD, it is important to start with a valid animal model of the disorder. This protocol describes the methodology required to conduct stress-enhanced fear learning (SEFL) experiments, a preclinical model of PTSD, in both rats and mice. SEFL was developed to recapitulate critical aspects of PTSD, including long-term sensitization of fear learning caused by an acute stressor. SEFL uses aspects of Pavlovian fear conditioning but produces a distinct and robust sensitized fear response far greater than normal conditional fear responses. The trauma procedure involves placing a rodent in a conditioning chamber and administering 15 unsignaled shocks randomly distributed over 90 minutes (for rat experiments; for mouse experiments, 10 unsignaled shocks randomly distributed over 60 minutes are used). On day 2, rodents are placed in a novel conditioning context where they receive a single shock; then, on day 3 they are placed back in the same context as on day 2 and tested for changes in freezing levels. Rodents that previously received the trauma display enhanced levels of freezing on the test day compared to those that received no shocks on the first day. Thus, with this model, a single highly stressful experience (the trauma) produces extreme fear of the stimuli associated with the traumatic event.

Here we describe the detailed methodology required to conduct stress-enhanced fear learning (SEFL) experiments, a preclinical model of post-traumatic stress disorder, in both rats and mice. The model utilizes aspects of Pavlovian fear conditioning and freezing as an index of enhanced fear in rodents.

Stress verstärkt Angst lernen, ein stabiles Nagetier Modell der posttraumatischen Belastungsstörung

Fear behaviors are important for survival, but disproportionately high levels of fear can increase the vulnerability for developing psychiatric disorders ...

Visual Classical Conditioning in Wood Ants

Several species of insects have become model systems for studying learning and memory formation. Although many studies focus on freely moving animals, studies implementing classical conditioning paradigms with harnessed insects have been important for investigating the exact cues that individuals learn and the neural mechanisms underlying learning and memory formation. Here we present a protocol for evoking visual associative learning in wood ants through classical conditioning. In this paradigm, ants are harnessed and presented with a visual cue (a blue cardboard), the conditional stimulus (CS), paired with an appetitive sugar reward, the unconditional stimulus (US). Ants perform a Maxilla-Labium Extension Reflex (MaLER), the unconditional response (UR), which can be used as a readout for learning. Training consists of 10 trials, separated by a 5-minute intertrial interval (ITI). Ants are also tested for memory retention 10 minutes or 1 hour after training. This protocol has the potential to allow researchers to analyze, in a precise and controlled manner, the details of visual memory formation and the neural basis of learning and memory formation in wood ants.

We present a protocol for the classical conditioning of harnessed ants that permits researchers to study visual learning and memory formation at a level of analysis not possible with freely moving individuals.

Visuelle Klassische Konditionierung in Waldameisen

Several species of insects have become model systems for studying learning and memory formation. Although many studies focus on freely moving animals, ...

Fear Incubation Using an Extended Fear-Conditioning Protocol for Rats

Emotional memory has been primarily studied with fear-conditioning paradigms. Fear conditioning is a form of learning through which individuals learn the relationships between aversive events and otherwise neutral stimuli. The most-widely utilized procedures for studying emotional memories entail fear conditioning in rats. In these tasks, the unconditioned stimulus (US) is a footshock presented once or several times across single or several sessions, and the conditioned response (CR) is freezing. In a version of these procedures, called cued fear conditioning, a tone (conditioned stimulus, CS) is paired with footshocks (US) during the training phase. During the first test, animals are exposed to the same context in which training took place, and freezing responses are tested in the absence of footshocks and tones (i.e., a context test). During the second test, freezing is measured when the context is changed (e.g., by manipulating the smell and walls of the experimental chamber) and the tone is presented in the absence of footshocks (i.e., a cue test). Most cued fear conditioning procedures entail few tone-shock pairings (e.g., 1-3 trials in a single session). There is a growing interest in less common versions involving an extensive number of pairings (i.e., overtraining) related to the long-lasting effect called fear incubation (i.e., fear responses increase over time without further exposure to aversive events or conditioned stimuli). Extended fear-conditioning tasks have been key to the understanding of fear incubation&#8217;s behavioral and neurobiological aspects, including its relationship with other psychological phenomena (e.g., post-traumatic stress disorder). Here, we describe an extended fear-conditioning protocol that produces overtraining and fear incubation in rats. This protocol entails a single training session with 25 tone-shock pairings (i.e., overtraining) and a comparison of conditioned freezing responses during context and cue tests 48 h (short-term) and 6 weeks (long-term) after training.

We describe an extended fear-conditioning protocol that produces overtraining and fear incubation in rats. This protocol entails a single training session with 25 tone-shock pairings (i.e., overtraining) and a comparison of conditioned freezing responses during context and cue tests 48 h (short-term) and 6 weeks (long-term) after training.

Angst-Inkubation mit einem erweiterten Angst-Konditionierungsprotokoll für Ratten

Emotional memory has been primarily studied with fear-conditioning paradigms. Fear conditioning is a form of learning through which individuals learn the ...