This work is supported by a Baylor University Research Council grant and by a research grant from the Epilepsy Foundation.

Die erwachsenen männlichen Mäusen im folgenden Experiment verwendet wurden, erzeugt und untergebracht in Baylor University bei einer Umgebungstemperatur von 22 ° C, mit einem 14 h Licht und 10 h Dunkelheit (20.00 bis 06.00 h) Tageszyklus. Die Mäuse wurden ad libitum Zugang zu Futter und Wasser. Alle Verfahren, um die Mäuse in Übereinstimmung mit den National Institutes of Health Leitlinien für die Pflege und Verwendung von Labortieren und dem Tier-Protokoll wurde von der Baylor University Animal Care und Verwenden Committee genehmigt. Die Mäuse wurden am späten Vormittag / frühen Nachmittag getestet. Für alle Verfahren unter der Prüfer sollte Handschuhe tragen, um Gerüche zu beseitigen und zu verhindern Kontamination. 1. Herstellung von Non-sozialen Gerüche <ol><li> Bereiten Sie frische nicht-sozialen Gerüche jeden Testtag. Hinweis: Der Prüfer sollte Handschuhe verwenden, um die Gerüche vorzubereiten. <ol><li> Füllen Sie 3 Zentrifugenröhrchen mit 10 ml Wasser. </li><li> Je 100 &amp; mgr; l der Bananenextrakt in eine 15 ml &amp; #160; konischen Röhrchen und 100 ul Mandelextrakt in ein zweites 15 ml konischen Röhrchen für eine 1: 100 Verdünnung. Hinweis: Die dritte 15-ml-Tube werden nur Wasser enthalten. </li></ol></li></ol> 2. Herstellung von Social Gerüche <ol><li> Bereiten Sie frische sozialen Gerüche am Morgen Tests auftreten wird. Erhalten soziale Gerüche aus zwei Käfige, die für mindestens drei Tage gereinigt wurden, und die gleiche Anzahl von Mäusen. Erhalten Sozial Gerüche aus Mäusen des gleichen Geschlechts wie der Testperson. Hinweis: Es handelt sich um bei Raumtemperatur gelagert werden und sollte innerhalb von 4-6 Stunden verwendet werden, um starke und konsistente Geruch zu halten. <ol><li> Ermitteln Sie, wie viele soziale Geruch Wattestäbchen erstellt werden müssen: jede Maus, die getestet wird mit drei Abstriche aus sozialen Geruch 1, und drei von sozialen Geruch 2 vorgestellt. </li><li> Bereiten sozialen Gerüche durch Abwischen ein Wattestäbchen im Zickzack über den Boden eines schmutzigen Käfig. Schütteln Sie überschüssiges Bettfesthalten an der Spitze. </li><li> Shop Tupfer in große Gläser mit Deckel oder in Bechergläsern mit Parafilm bedeckt. Bestimmen Sie einen Behälter für die soziale Geruch 1 und einen zweiten für die soziale Geruch 2. </li></ol></li><li> Pflegen Geruchsquelle Käfige außerhalb der experimentellen Testraum. </li></ol> 3. Akklimatisierung des Test-Maus <ol><li> Setzen Sie auf Handschuhe und wiegen Sie die Maus zu testenden und individuell in einem sauberen Käfig mit einem Draht Deckel auf der Oberseite und der Wasserflasche entfernt. </li><li> Lassen Sie die Maus an den neuen Käfig für 45 min in einem anderen Raum als der Testraum akklimatisieren. Stellen Sie sicher, dass es keine ungewöhnliche Gerüche oder Geräusche im Raum. </li><li> Als Test dauert etwa 45 min, taumeln die Akklimatisierung Zeiten, indem die nächste Maus, um in einen sauberen Käfig getestet, um direkt vor die Prüfung der aktuellen Maus akklimatisieren werden. </li></ol> 4. Herstellung von den Testraum <ol><li> Zeigen frischen Wattestäbchen, Labor-Band, die Flaschen mit sozialen Geruch Tupfer, die Rohre von non-sozialen Gerüche, eine Mikropipette, zwei Timer und eine Abfallflasche mit einem Deckel in den Testraum. </li><li> Stellen Sie sicher, dass die Testraum ist weitgehend frei von Gerüchen, laute Geräusche und helles Licht. Tragen Sie keine Duft Kölnisch Wasser oder Lotionen. Hinweis: Die Beobachter sollten nicht Kaugummi oder Bonbons während der Testphase und sollte versuchen, ihre Exposition gegenüber starken Geruch Lebensmitteln zu begrenzen. </li></ol> 5. Testverfahren <ol><li> Bringen Sie die Maus gewöhnt in seinem Käfig in den Testraum und platzieren Sie den Käfig auf der Theke in einer Höhe, die bequem für die Anzeige im Sitzen ist. </li><li> Präsentieren Sie jedem Geruch 3 mal hintereinander für 2 Minuten jedes Mal. Verwenden Sie die Darstellung um, die in Tabelle 1 zu finden sind. <ol><li> Je 50 ul der Geruchsstoff in dem Duft auf, um die Spitze des Wattebausch getestet und neu Kappe die Geruchsquelle. Heben Sie den Draht Spitze des Käfigs und legen Sie die unscented, Holz Ende der Tupfer in das Wasser Flaschenöffnung, pushing die Länge durch die von der Unterseite des Drahtes an die Spitze. Berühren Sie die Wattestäbchen, um alles, einschließlich Finger, so dass Geruchskontamination vermieden wird. </li><li> Fahren Sie mit dem Tupfer durch den Wasserflaschenöffnung zu ziehen, bis etwa 1 Zoll des Baumwoll Ende nach unten hängt in den Käfig. Befestigen Sie das Ende des Holz Tupfer auf die Seite des Käfigs mit Labor Band. Hinweis: Achten Sie darauf, den Deckel wieder auf den Käfig fallen zu lassen, was ein lautes Geräusch, das den Test der Maus stören würden. </li><li> Sofort beginnen Timing für 2 min. Verwenden Sie einen Timer, um die 2 Minuten Präsentationszeit. Notieren Sie sich die kumulative Zeit verbrachte Sniffing den Geruch während der 2 Minuten-Studie. Graf Schnüffeln das Verhalten des Tieres mit einem separaten Stoppuhr. Aktive Schnüffeln wird definiert, wenn der Nase des Tieres in Richtung auf die Baumwollspitze mit seiner Nase ~ 2 cm oder näher an der Spitze ausgerichtet ist. </li><li> Nach dem Abschluss der jeden Versuch, heben Sie den Draht Deckel und entfernen Sie die Wattestäbchen von unten ziehen es durch die unterseite des Deckels (dies vermeidet Geruchsbelastung durch die Verhinderung der Baumwollspitze berührt den Draht). Verschließen Sie den verwendeten Tupfer in einem 500 ml Abfallflasche. </li><li> Bereiten Sie und legen Sie die nächste Tupfer und beginnen Timing. Lassen Sie ca. 1 min zwischen jeder inter-trial-Intervall. Fortfahren, bis jeder Duft wurde drei Mal in Folge präsentiert, für 2 Minuten jedes Mal. </li></ol></li><li> Sobald die Maus hat alle Prüfungen abgeschlossen ist, leeren Sie den Abfallbehälter, die die verwendeten Wattestäbchen in einem Papierkorb Außenseite kann der Testraum. Rückflug die Maus, um seinen eigenen Käfig. </li></ol> 6. Analyse <ol><li> Analysieren Sie Daten für jede Gruppe pro Geruch Zustand mit Zwei-Wege-ANOVA, gefolgt von Newman-Keuls oder eine ähnliche Post-hoc-Test, um festzustellen, erhebliche Habituation (weniger Zeit Sniffing aufeinanderfolgenden gleichen Gerüche) und dishabitutation (mehr Zeit Sniffing einen neuen Geruch). </li></ol>

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A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Olfactory+dysfunction+correlates+with+amyloid-beta+burden+in+an+Alzheimer's+disease+mouse+model.">Olfactory dysfunction correlates with amyloid-beta burden in an Alzheimer's disease mouse model.</a> J Neurosci. 30, 505-514 (2010).</li><li>Fadool, D. A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Kv1.3+channel+gene-targeted+deletion+produces+'Super-Smeller+Mice'+with+altered+glomeruli,+interacting+scaffolding+proteins,+and+biophysics.">Kv1.3 channel gene-targeted deletion produces 'Super-Smeller Mice' with altered glomeruli, interacting scaffolding proteins, and biophysics.</a> Neuron. 41, 389-404 (2004).</li><li>Arakawa, H., Arakawa, K., Blanchard, D. C., Blanchard, R. 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N., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Social+amnesia+in+mice+lacking+the+oxytocin+gene.">Social amnesia in mice lacking the oxytocin gene.</a> Nat Genet. 25, 284-288 (2000).</li><li>Yang, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Low+sociability+in+BTBR+T+tf/J+mice+is+independent+of+partner+strain.">Low sociability in BTBR T+tf/J mice is independent of partner strain.</a> Physiol Behav. 107, 649-662 (2012).</li><li>Best, J. D., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Non-associative+learning+in+larval+zebrafish.">Non-associative learning in larval zebrafish.</a> Neuropsychopharmacology. 33, 1206-1215 (2008).</li><li>Wolman, M. A., Jain, R. 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The authors have nothing to disclose.

Die Ergebnisse in diesem Papier optimal sind für Mäuse. Die Mäuse zeigen eine starke Reaktion auf eine neuartige Geruchs, dann schnell zu gewöhnen. Einer der wichtigsten Schritte bei diesem Verfahren ist die Herstellung der Gerüche. Der Prüfer muss sehr vorsichtig sein, um die Gerüche voneinander zu isolieren, um eine Kontamination zu verhindern. Ein weiterer wichtiger Aspekt ist die Präsentation der Wattestäbchen. Der Prüfer muss die beste Lage des Baumwollspitze zu beheben, so dass die Maus können Sie die S...

Mäuse sind abhängig von Geruchssinn für die Navigation in neuen Umgebungen, die Suche nach Nahrung, zum Erkennen von anderen Personen und Sexualverhalten 1-3. Es ist wichtig, dass die Ermittler festzustellen, ob Versuchstieren eine funktionierende Geruchssinn vor der Verabreichung von Verhaltenstests, die Lebensmittel, die soziale Interaktion oder Gerüche bestimmt, um eine Antwort von der Maus zu entlocken einzubeziehen. Anosmie oder die Unfähigkeit, zwischen verschiedenen Gerüchen zu unterscheiden, könnte in falschen Positiven oder Negativen in einer Vielzahl von Verhaltensparadigmen führen, so einer Maus Fähigkeit zu erkennen und zu unterscheiden Gerüche sollte eingerichtet werden, bevor andere Arten von Verhaltenstests durchgeführt werden. Das olfaktorische Gewöhnung / Dishabituation Test wurde zuerst in den 1980er Jahren 4 beschrieben. Der Test wurde für die Verwendung in Mäusen durch Drs angepasst. Mu Yang und Jacqueline Crawley 5. Dies ist eine einfache und kostengünstige Test, dass der Prüfer ESTA erlaubtblish, die eine Maus zu erkennen und zu unterscheiden zwischen Gerüchen. Neben der Prüfung Geruchssinn, dieser Test ermöglicht dem Ermittler, um die allgemeine Verhalten der Maus zu beobachten und sollten frühzeitig in einem Testschema durchgeführt werden. Qualitative Beobachtungen bezüglich der Maus die Fortbewegung, Anzeichen von Angst, das Aktivitätsniveau, und die Reaktion auf gesellschaftliche Gerüchen gegenüber Nahrungsmittelgerüche können neue Bereiche, in denen Tests durchgeführt werden konnte signalisieren. In diesem Test, Wattestäbchen in verschiedene Gerüche getaucht werden einer Maus dreimal in einer Zeile dargestellt. Bei jedem wiederholten Präsentation eines Geruchs wird die Maus an den Wattestäbchen gewöhnen, verbringen weniger Zeit damit es mit jeder weiteren Präsentation. Wenn ein neuer Geruch präsentiert wird, tritt Entwöhnung und eine typische Maus wird mehr Zeit für die Untersuchung der Tupfer, was darauf hinweist, dass es zwischen den aktuellen und früheren Gerüche 5 unterscheiden. Dieser Test wird auf eine Maus zu einem Zeitpunkt verabreicht und umfasst eine 45 min nachlimation Periode, gefolgt von 45 Minuten der Prüfung. Obwohl dieser Test ist einfach durchzuführen kann es verwendet werden, um anspruchsvolle Fragen über die Maus olfaktorischen Systems zu untersuchen. Weitere beliebte Tests des Geruchssinns, wie beispielsweise die Lebensmittel begraben Test, einfach stellen die Anwesenheit oder Abwesenheit von Anosmie. Allerdings ermöglicht der Geruchs Diskriminierung und Gewöhnungstest der Forscher, um festzustellen, dass eine Maus hat nicht nur die Fähigkeit, Gerüche zu erkennen, sondern können zwischen verschiedenen Gerüchen zu unterscheiden. Das Muster der Gewöhnung und Dishabituation verwendet wurde, um festzustellen, ob neue Mutanten können zwischen 6,7 Gerüchen zu unterscheiden. In einer überraschenden Studie Fadool und Kollegen verwendet komplexe Mischungen von Gerüchen, dass Mäuse mit Gen-Targeting Löschung der Kv1.3 Kanal zeigen, sind &quot;Super smellers&quot;, die sehr ähnlich Gerüche besser als normale Mäuse 8 unterscheiden kann. Bei der Prüfung eines neuen Knockout-Mausmodell es sinnvoll ist establish Gegenwart von normalem Verhalten für Grund sensorischen Aufgaben. Wenn früh in einem Testprozeduren durchgeführt, der Geruch von Diskriminierung und Gewöhnung Test gibt ein Ermittler die Möglichkeit, ungewöhnliche Verhaltensweisen zu beobachten. Diese Beobachtungen könnten falsch positive oder negative Ergebnisse in nachfolgenden Tests, zuzurechnen Eigenschaften des mutierten verwechseln sind zu vermeiden. Als Forscher weiterhin Sozialverhalten die Notwendigkeit, grundlegende Riechfunktion überprüfen untersuchen immer wichtiger. Neben der Prüfung Geruchssinn in Mutantenlinien kann dieser Test verwendet werden, um spezifische Fragen, wie beispielsweise, ob eine pharmakologische Behandlung Reaktion eines Tieres auf soziale Geruch Reize gezielt erhöht oder wenn ihre Reaktion erhöht sich auf alle Geruchsreize ansprechen.

<table border="0" cellpadding="0" cellspacing="0">
	<tbody>
		<tr>
			<td>Mouse cage</td>
			<td>Allentown</td>
			<td>Standard mouse cage</td>
			<td></td>
		</tr>
		<tr>
			<td rowspan="2">Wire lid</td>
			<td rowspan="2">Allentown</td>
			<td>BCU Mouse WBL 2500</td>
			<td rowspan="2"></td>
		</tr>
		<tr>
			<td></td>
		</tr>
		<tr>
			<td>Bedding</td>
			<td>Harlan</td>
			<td>7090 Sani-Chips</td>
			<td></td>
		</tr>
		<tr>
			<td>Cotton swabs</td>
			<td>VWR</td>
			<td>89031-270</td>
			<td>6&#8221; wooden handle</td>
		</tr>
		<tr>
			<td>Banana extract</td>
			<td>McCormick</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Almond extract</td>
			<td>McCormick</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Laboratory tape</td>
			<td>VWR</td>
			<td>89098-062</td>
			<td></td>
		</tr>
		<tr>
			<td>Stop watch</td>
			<td>VWR</td>
			<td>62374-000</td>
			<td></td>
		</tr>
		<tr>
			<td>Nitrile gloves</td>
			<td>VWR</td>
			<td>82026</td>
			<td></td>
		</tr>
		<tr>
			<td>Timing device</td>
			<td>VWR</td>
			<td>61161-350</td>
			<td></td>
		</tr>
		<tr>
			<td>15 mL conical tubes</td>
			<td>VWR</td>
			<td>89003-294</td>
			<td></td>
		</tr>
		<tr>
			<td>2 L beakers&#160;</td>
			<td>Pyrex</td>
			<td align="right">1003</td>
			<td></td>
		</tr>
		<tr>
			<td>Parafilm</td>
			<td>Parafilm</td>
			<td>PM-992</td>
			<td>4&#8221; x 250&#8217;</td>
		</tr>
		<tr>
			<td>1 L bottle with cap</td>
			<td>VWR</td>
			<td>89000-240</td>
			<td></td>
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</table>

testing for odor discrimination and habituation in mice

This video demonstrates a technique to establish the presence of a normally functioning olfactory system in a mouse. The test helps determine whether the mouse can discriminate between non-social odors and social odors, whether the mouse habituates to a repeatedly presented odor, and whether the mouse demonstrates dishabituation when presented with a novel odor. Since many social behavior tests measure the experimental animal&#8217;s response to a familiar or novel mouse, false positives can be avoided by establishing that the animals can detect and discriminate between social odors. There are similar considerations in learning tests such as fear conditioning that use odor to create a novel environment or olfactory cues as an associative stimulus. Deficits in the olfactory system would impair the ability to distinguish between contexts and to form an association with an olfactory cue during fear conditioning. 
In the odor habitation/dishabituation test, the mouse is repeatedly presented with several odors. Each odor is presented three times for two minutes. The investigator records the sniffing time directed towards the odor as the measurement of olfactory responsiveness. A typical mouse shows a decrease in response to the odor over repeated presentations (habituation). The experimenter then presents a novel odor that elicits increased sniffing towards the new odor (dishabituation). After repeated presentation of the novel odor the animal again shows habituation. This protocol involves the presentation of water, two or more non-social odors, and two social odors. In addition to reducing experimental confounds, this test can provide information on the function of the olfactory systems of new knockout, knock-in, and conditional knockout mouse lines.

Nach dem beschriebenen Protokoll wurden die Ergebnisse unter Verwendung von zehn Erwachsenen (postnatalen Tag 90 bis 120) männliche C57BL / 6J Mäusen und acht männlichen FVB mixed-Hintergrund Stamm (postnatalen Tag 50-70) aufgezeichnet. Die Prüfung erfolgte durch einen einzigen Untersucher über mehrere Tage durchgeführt. Jeder Punkt auf dem Graphen repräsentiert die durchschnittliche Verweilzeit der Untersuchung eines Geruch. Wenn das Wattestäbchen in Wasser getaucht wird vorgelegt, ...

Watch this Scientific Journal Video about Testing for Odor Discrimination and Habituation in Mice at JoVE.com

Testing for Odor Discrimination and Habituation in Mice

This manuscript describes a protocol to examine the olfactory system of rodents. The olfactory habituation/dishabituation test will allow investigators to determine whether a mouse habituates to a repeatedly presented odor and whether the mouse demonstrates dishabituation when presented a novel odor.

Die Prüfung auf Geruch Diskriminierung und Habituation in Mice

This video demonstrates a technique to establish the presence of a normally functioning olfactory system in a mouse. The test helps determine whether the ...

testing-for-odor-discrimination-and-habituation-in-mice

Research

JoVE Journal

Behavior

17.5K Aufrufe.  Baylor University. This manuscript describes a protocol to examine the olfactory system of rodents. The olfactory habituation/dishabituation test will allow investigators to determine whether a mouse habituates to a repeatedly presented odor and whether the mouse demonstrates dishabituation when presented a novel odor. 

Serie: Testing for Odor Discrimination and Habituation 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

Barnes Maze Testing Strategies with Small and Large Rodent Models

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

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

Barnes Maze Teststrategien mit kleinen und großen Nager-Modelle

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

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 ...

Morris Water Maze Test: Optimization for Mouse Strain and Testing Environment

The Morris water maze (MWM) is a commonly used task to assess hippocampal-dependent spatial learning and memory in transgenic mouse models of disease, including neurocognitive disorders such as Alzheimer&#8217;s disease. However, the background strain of the mouse model used can have a substantial effect on the observed behavioral phenotype, with some strains exhibiting superior learning ability relative to others. To ensure differences between transgene negative and transgene positive mice can be detected, identification of a training procedure sensitive to the background strain is essential. Failure to tailor the MWM protocol to the background strain of the mouse model may lead to under- or over- training, thereby masking group differences in probe trials. Here, a MWM protocol tailored for use with the F1 FVB/N x 129S6 background is described. This is a frequently used background strain to study the age-dependent effects of mutant P301L tau (rTg(TauP301L)4510 mice) on the memory deficits associated with Alzheimer&#8217;s disease. Also described is a strategy to re-optimize, as dictated by the particular testing environment utilized.

This manuscript describes a Morris water maze (MWM) protocol tailored for use with a commonly used mouse model of Alzheimer's disease. The MWM is widely used in transgenic mouse models. Implementation of a procedure sensitive to the background strain of the mouse model is essential for detecting group differences.

Morris Wasser-Labyrinth-Test: Optimierung für Mausstamm und Testumgebung

 The Morris water maze (MWM) is a commonly used task to assess hippocampal-dependent spatial learning and memory in transgenic mouse models of disease, ...

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 ...

Simple and Computer-assisted Olfactory Testing for Mice

Olfaction is highly conserved among species and is required for reproduction and survival.
In humans, olfaction is also one of the senses that is affected with aging and is a strong predictor of neurodegenerative diseases. Thus, olfaction testing is used as a non-invasive diagnostic method to detect neurological deficits early on. In order to understand the mechanisms underlying olfactory network susceptibility, olfactory research in rodents has gained momentum in the past decade.
Here, we present a very simple, time efficient and reproducible olfactory testing method of innate odor perception and sensitivity in mice without the need of any prior food or water restriction. The tests are performed in a familiar environment to the mice, require only the scents and a 2 min session of odorant exposure. The analysis is performed, post-hoc, using computer-assisted commands on ImageJ and can be, therefore, carried out from start to end by one researcher.
This protocol does not require any special hardware or setup and is indicated for any laboratory interested in testing olfactory perception and sensitivity.

We present a simple and unbiased olfactory test in mice. With this protocol olfactory discrimination, preference, avoidance and sensitivity to a novel odor as compared to water can be assessed in single behavioral sessions. This method is indicated for a single experimenter and analysis is based on computer-assisted video processing.

Einfache und Computergestützte Riechtests für Mäuse

Olfaction is highly conserved among species and is required for reproduction and survival.
In humans, olfaction is also one of the senses that is affected ...

Testing for Metacognitive Responding Using an Odor-based Delayed Match-to-Sample Test in Rats

Metamemory involves the cognitive ability to assess the strength of one&#39;s memories. To explore the possibility of metamemory in non-human animals, numerous behavioral tasks have been created, many of which utilize an option to decline memory tests. To assess metamemory in rats, we utilized this decline-test option paradigm by adapting previous visual delayed-match-to-sample tests (DMTS)1,2 developed for primate species to an odor-based test suitable for rodents. First,&#160;rats are given a sample to remember by digging in a cup of scented sand. After a delay, the rat is presented with four distinctly scented cups, one of which contains the identical scent experienced during the sample; if this matching cup is selected, then the rat obtains a preferred, larger reward. Selection of any of the other three non-matching sand-filled scented cups results in no reward. Retention intervals are individually titrated such that subjects perform between 40 and 70% correct, therefore ensuring rats sometimes remember and sometimes forget the sample. Here, the operational definition of metamemory is the ability to distinguish between the presence and absence of memory through behavioral responding. Towards this end, on two-thirds of trials, a decline option is presented in addition to the four choice cups (choice trials). If the decline-test option- an unscented colored sand cup, is selected, the subject receives a smaller less-preferred reward and avoids the memory test. On the remaining third of trials, the decline-test option is not available (forced trials), causing subjects to guess the correct cup when the sample is forgotten. On choice tests, subjects that know when they remember should select the decline option when memory is weak rather than take the test and choose incorrectly. Therefore, significantly higher performance on chosen tests as compared to forced memory tests is indicative of the adaptive use of the decline-test response and metacognitive responding.

This protocol describes a method for investigating the possibility of metamemory, or memory awareness, in rodents. The odor-based delayed-matching-to-sample paradigm is a novel, ecologically-relevant behavioral test useful for determining the extent to which rodents can adaptively respond based on cognitively monitoring the strength of their memory states.

Für die metakognitiven Reaktion mit einem Geruch-basierte Tests verzögert Match Beispieltest bei Ratten

Metamemory involves the cognitive ability to assess the strength of one&#39;s memories. To explore the possibility of metamemory in non-human animals, ...

Measuring Active and Passive Tameness Separately in Mice

Domesticated animals such as dogs and laboratory mice show a high level of tameness, which is important for humans to handle them easily. Tameness has two behavioral components: a reluctance to avoid humans (passive tameness) and a motivation to approach humans (active tameness). To quantify these components in mice, we previously developed behavioral tests for active tameness, passive tameness, and the willingness to stay on a human hand, each designed to be completed within 3 min. The data obtained were used for selective breeding, with a large number of mice analyzed per generation. The active tameness test measures the movement of the mouse toward a human hand and the contact it engages in. The passive tameness test measures the duration of time that a mouse tolerates human touch. In the stay-on-hand test, a mouse is placed on a human hand and touched slowly using the thumb of that hand; the duration of time that the animal remains on the hand is measured. Here, we describe the test set-up and apparatus, explain the procedures, and discuss the appropriate data analysis. Finally, we explain how to interpret the results.

Tameness in animals includes a reduction of avoidance responses to humans (passive tameness) and an increase in active approaches to humans (active tameness). Here, we describe detailed protocols for three behavioral tests (active tameness, passive tameness, and stay-on-hand tests) to separately measure the active and passive tameness of mice.

Messung der aktiven und passiven Zahmheit separat bei Mäusen

Domesticated animals such as dogs and laboratory mice show a high level of tameness, which is important for humans to handle them easily. Tameness has two ...

A Wind Tunnel for Odor Mediated Insect Behavioural Assays

Olfaction is the most important sensory mechanism by which many insects interact with their environment and a wind tunnel is an excellent tool to study insect chemical ecology. Insects can locate point sources in a three-dimensional environment through the sensory interaction and sophisticated behavior. The quantification of this behavior is a key element in the development of new tools for pest control and decision support. A wind tunnel with a suitable flight section with laminar air flow, visual cues for in-flight feedback and a variety of options for the application of odors can be used to measure complex behaviour which subsequently may allow the identification of attractive or repellent odors, insect flight characteristics, visual-odor interactions and interactions between attractants and odors lingering as background odors in the environment. A wind tunnel holds the advantage of studying the odor mediated behavioural repertoire of an insect in a laboratory setting. Behavioural measures in a controlled setting provide the link between the insect physiology and field application. A wind tunnel must be a flexible tool and should easily support the changes to setup and hardware to fit different research questions. The major disadvantage to the wind tunnel setup described here, is the clean odor background which necessitates special attention when developing a synthetic volatile blend for field application.

Here, we describe the construction and use of a wind tunnel for odor mediated behavioural assays with insects. The wind tunnel design facilitates the release of odor sources by several methods, with and without visual stimuli. Wind tunnel experiments are important methods to identify behaviorally active volatile chemicals.

Ein Windkanal für Geruch vermittelt Insekt Behavioral Assays

Olfaction is the most important sensory mechanism by which many insects interact with their environment and a wind tunnel is an excellent tool to study ...

Novel Object Recognition and Object Location Behavioral Testing in Mice on a Budget

Ethologically relevant behavioral testing is a critical component of any study that uses mouse models to study the cognitive effects of various physiological or pathological changes. The object location task (OLT) and the novel object recognition task (NORT) are two effective behavioral tasks commonly used to reveal the function and relative health of specific brain regions involved in memory. While both of these tests exploit the inherent preference of mice for the novelty to reveal memory for previously encountered objects, the OLT primarily evaluates spatial learning, which relies heavily on hippocampal activity. The NORT, in contrast, evaluates non-spatial learning of object identity, which relies on multiple brain regions. Both tasks require an open-field-testing arena, objects with equivalent intrinsic value to mice, appropriate environmental cues, and video recording equipment and the software. Commercially available systems, while convenient, can be costly. This manuscript details a simple, cost-effective method for building the arenas and setting up the equipment necessary to perform the OLT and NORT. Furthermore, the manuscript describes an efficient testing protocol that incorporates both OLT and NORT and provides typical methods for data acquisition and analysis, as well as representative results. Successful completion of these tests can provide valuable insight into the memory function of various mouse model systems and appraise the underlying neural regions that support these functions.

Here we provide a protocol which includes comprehensive instructions for the economical establishment of murine object location and novel object recognition behavioral testing, including the design, cost, and construction of required equipment as well as execution of behavioral testing, data collection, and analysis.

Methods Article

Die Prüfung auf Geruch Diskriminierung und Habituation in Mice