Name: novel object recognition test for the investigation of learning and memory in mice
Uploaded: 2017-08-30T13:00:00
Description: Watch this Scientific Journal Video about Novel Object Recognition Test for the Investigation of Learning and Memory in Mice at JoVE.com

Work cited and previously published by the author was supported by a grant from the National Institute of Mental Health (MH088480). The author would like to thank her former mentor, Dr. James O'Donnell for his support in that project. This publication is supported by a grant from the National Institute of Health (T32 DA007135).

All procedures performed here were submitted to and approved by the Animal Care and Use committee and were conducted following NIH guidelines.
1. Object Selection and Experimental Setup
<ol>
	<li>Select objects that are different enough to be easily discriminated by mice, but have a similar degree of complexity (texture, shape, color patterning and brightness, etc.) in order to minimize any potential induced object preference that may bias the results (see Ennaceur 2010 for ...

<ol>
	<li>Ennaceur, A., Meliani, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+new+one-trial+test+for+neurobiological+studies+of+memory+in+rats+III.+Spatial+vs.+non-spatial+working+memory.">A new one-trial test for neurobiological studies of memory in rats III. Spatial vs. non-spatial working memory.</a> Behav. Brain Res. 51 (1), 83-92 (1988).</li><li>Akkerman, S., 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=Object+recognition+testing:+methodological+considerations+on+exploration+and+discrimination+measures.">Object recognition testing: methodological considerations on exploration and discrimination measures.</a> Behav. Brain Res. 232 (2), 335-347 (2012).</li><li>Antunes, M., Biala, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+novel+object+recognition+memory:+neurobiology,+test+procedure,+and+its+modifications.">The novel object recognition memory: neurobiology, test procedure, and its modifications.</a> Cogn. Process. 13 (2), 93-110 (2012).</li><li>Leger, 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=Object+recognition+test+in+mice.">Object recognition test in mice.</a> Nat. Protoc. 8 (12), 2531-2537 (2013).</li><li>van Goethem, N. P., 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=Object+recognition+testing:+Rodent+species,+strains,+housing+conditions,+and+estrous+cycle.">Object recognition testing: Rodent species, strains, housing conditions, and estrous cycle.</a> Behav. Brain Res. 232 (2), 323-334 (2012).</li><li>Lueptow, L. M., Zhang, C. -. G., O'Donnell, J. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cyclic+GMP-mediated+memory+enhancement+in+the+object+recognition+test+by+inhibitors+of+phosphodiesterase-2+in+mice.">Cyclic GMP-mediated memory enhancement in the object recognition test by inhibitors of phosphodiesterase-2 in mice.</a> Psychopharmacology (Berl). , (2015).</li><li>Ennaceur, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=One-trial+object+recognition+in+rats+and+mice:+Methodological+and+theoretical+issues.">One-trial object recognition in rats and mice: Methodological and theoretical issues.</a> Behav. Brain Res. 215 (2), 244-254 (2010).</li><li>Berlyne, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Novelty+and+curiosity+as+determinants+of+exploratory+behavior.">Novelty and curiosity as determinants of exploratory behavior.</a> Br. J. Psychol. 41 (1-2), 68-80 (1950).</li><li>Ennaceur, A., Delacour, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+new+one-trial+test+for+neurobiological+studies+of+memory+in+rats+I.+Behavioral-data.">A new one-trial test for neurobiological studies of memory in rats I. Behavioral-data.</a> Behav. 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Pharmacol. 436 (1-2), 83-87 (2002).</li><li>Bertaina-Anglade, V., Enjuanes, E., Morillon, D., Drieu la Rochelle, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+object+recognition+task+in+rats+and+mice:+A+simple+and+rapid+model+in+safety+pharmacology+to+detect+amnesic+properties+of+a+new+chemical+entity.">The object recognition task in rats and mice: A simple and rapid model in safety pharmacology to detect amnesic properties of a new chemical entity.</a> J. Pharmacol. Toxicol. 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Brain Res. 285, 140-157 (2015).</li><li>Balderas, I., Moreno-Castilla, P., Bermudez-Rattoni, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dopamine+D1+receptor+activity+modulates+object+recognition+memory+consolidation+in+the+perirhinal+cortex+but+not+in+the+hippocampus.">Dopamine D1 receptor activity modulates object recognition memory consolidation in the perirhinal cortex but not in the hippocampus.</a> Hippocampus. 23 (10), 873-878 (2013).</li><li>Akkerman, S., Blokland, A., Prickaerts, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mind+the+gap:+delayed+manifestation+of+long-term+object+memory+improvement+by+phosphodiesterase+inhibitors.">Mind the gap: delayed manifestation of long-term object memory improvement by phosphodiesterase inhibitors.</a> Neurobiol. Learn. 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Brain Res. 236 (1), 16-22 (2013).</li><li>Deacon, R. M. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Housing,+husbandry+and+handling+of+rodents+for+behavioral+experiments.">Housing, husbandry and handling of rodents for behavioral experiments.</a> Nat. Protoc. 1 (2), 936-946 (2006).</li><li>&#350;&#305;k, A., van Nieuwehuyzen, P., Prickaerts, J., Blokland, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Performance+of+different+mouse+strains+in+an+object+recognition+task.">Performance of different mouse strains in an object recognition task.</a> Behav. Brain Res. 147 (1-2), 49-54 (2003).</li><li>Prut, L., Belzung, C., Rabelias, U. 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The ORT is an efficient and flexible method for studying learning and memory in mice. When setting up an experiment, it is important to consider a number of variables that may affect the outcome. As discussed in the representative results, the strain of mouse will affect both exploration time and retention interval. A decrease in exploration time may skew or mask results in an absolute discrimination analysis2,3,5,<su...

The object recognition test (ORT), also known as the novel object recognition test (NOR), is a relatively fast and efficient means for testing different phases of learning and memory in mice. It was originally described by Ennaceur and Delacour in 1988 and used primarily in rats1; however, since then, it has been successfully adapted for use in mice2,3,4,5,6,7. The test relies on as few as three sessions: one habituation session, one training session, and one test session. Training simply involves visual exploration of two identical objects, while the test session involves replacing one of the previously explored objects with a novel object. Because rodents have an innate preference for novelty, a rodent that remembers the familiar object will spend more time exploring the novel object7,8,9.
The main advantage of the ORT over other rodent memory tests is that it relies on rodents&#39; natural proclivity for exploring novelty8. Therefore, there is no need for numerous training sessions or any positive or negative reinforcement to motivate behavior. This means that the ORT is much less stressful, relative to other tests10,11,12,13,14,15, and requires significantly less time to run than other commonly used memory tests, such as the Morris water maze or Barnes maze, which both can take up to a week or longer. Consequently, the conditions of the ORT more closely resemble those used in studying human cognition, increasing the ecological validity of the test over many other rodent memory tests. Similarly, because ORT is a simple visual recall task, it has been successfully adapted for use in numerous species, including humans and non-human primates, to assess different inter-species aspects of declarative memory 2,16,17. Finally, the ORT can be easily modified to examine different phases of learning and memory (i.e., acquisition, consolidation, or recall), to assess different types of memory (e.g., spatial memory), or to assess different retention intervals (i.e., short-term vs long-term memory).
The versatility of the ORT provides a platform for innumerable research applications. Studies can make use of pharmacologic agents to either disrupt or enhance memory. Varying the time of drug administration before or after training, or prior to testing can hint at the underlying neural mechanisms that lead to disrupted or enhanced memory6,18,19,20. In a similar way, optogenetic technology can be used at these same various time points to look at the neural activation/inhibition that contributes to the different phases of learning and memory. The ORT is also appropriate for assessing differences in transgenic animals, in lesion studies, or in neurodegenerative models or in aging studies21,22,23,24,25,26,27,28. The time between training and testing, known as the retention interval, can be altered to assess any of these changes on short- and long- term memory26. Ultimately, the ORT can be used as a tool to study pharmacological, genetic, and neurological changes to learning and memory, or these tools can be used to study the basis of learning and memory in the ORT.

<table border="0" cellpadding="0" cellspacing="0" width="805">
	<tbody>
		<tr height="84">
			<td height="84" style="height:84px;width:216px;">Open Field Box</td>
			<td style="width:71px;">Panlab/Harvard Apparatus</td>
			<td style="width:119px;">LE800SC</td>
			<td style="width:399px;">Available in grey, white, or black</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">ANY-maze</td>
			<td style="width:71px;">Stoelting Co.</td>
			<td style="width:119px;">60000</td>
			<td style="width:399px;">Behavior tracking system</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">EthoVisionXT 12</td>
			<td style="width:71px;">Noldus</td>
			<td style="width:119px;"></td>
			<td style="width:399px;">Behavior tracking system; requires 3 point tracking</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Video Camera</td>
			<td style="width:71px;">Any</td>
			<td style="width:119px;"></td>
			<td style="width:399px;">Video camera should be mounted directly overhead of the apparatus</td>
		</tr>
		<tr height="63">
			<td height="63" style="height:63px;width:216px;">70% Ethanol&#160;</td>
			<td style="width:71px;">Fisher Scientific</td>
			<td style="width:119px;">BP2818-4</td>
			<td style="width:399px;">Prior to starting testing and in between trials, each object should be carefully cleaned. The floor and walls of the apparatus should also be cleaned.&#160;</td>
		</tr>
	</tbody>
</table>

novel object recognition test for the investigation of learning and memory in mice

The object recognition test (ORT) is a commonly used behavioral assay for the investigation of various aspects of learning and memory in mice. The ORT is fairly simple and can be completed over 3 days: habituation day, training day, and testing day. During training, the mouse is allowed to explore 2 identical objects. On test day, one of the training objects is replaced with a novel object. Because mice have an innate preference for novelty, if the mouse recognizes the familiar object, it will spend most of its time at the novel object. Due to this innate preference, there is no need for positive or negative reinforcement or long training schedules. Additionally, the ORT can also be modified for numerous applications. The retention interval can be shortened to examine short-term memory, or lengthened to probe long-term memory. Pharmacological intervention can be used at various times prior to training, after training, or prior to recall to investigate different phases of learning (i.e., acquisition, early or late consolidation, or recall). Overall, the ORT is a relatively low-stress, efficient test for memory in mice, and is appropriate for the detection of neuropsychological changes following pharmacological, biological, or genetic manipulations.

A general experimental setup for the ORT is shown in Figure 2. On habituation day (T0) mice are placed in the empty arena for 5 min. Twenty-four hours later, mice are placed back in the chamber with 2 identical objects and allowed to freely explore for up to 10 min (T1). On testing day (T2), the mice are again placed in the arena, but with one familiar object and one novel object, and allowed to explore for up to 10 min. The retention interval, the time betwe...

Watch this Scientific Journal Video about Novel Object Recognition Test for the Investigation of Learning and Memory in Mice at JoVE.com

Novel Object Recognition Test for the Investigation of Learning and Memory in Mice

The object recognition test (ORT) is a simple and efficient assay for evaluating learning and memory in mice. The methodology is described below.

The object recognition test (ORT) is a commonly used behavioral assay for the investigation of various aspects of learning and memory in mice. The ORT is ...

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

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, ...

Novel Object Exploration as a Potential Assay for Higher Order Repetitive Behaviors in Mice

Restricted, repetitive behaviors (RRBs) are a core feature of autism spectrum disorder (ASD) and disrupt the lives of affected individuals.  RRBs are ...

The 4 Mountains Test: A Short Test of Spatial Memory with High Sensitivity for the Diagnosis of Pre-dementia Alzheimer's Disease

This protocol describes the administration of the 4 Mountains Test (4MT), a short test of spatial memory, in which memory for the topographical layout of ...

The Rodent Psychomotor Vigilance Test (rPVT): A Method for Assessing Neurobehavioral Performance in Rats and Mice

The Rodent Psychomotor Vigilance Test rPVT: A Method for Assessing Neurobehavioral Performance in Rats and Mice

The human Psychomotor Vigilance Test (PVT) is a widely used procedure for measuring changes in fatigue and sustained attention. The present article ...

A Real-world What-Where-When Memory Test

A Real-world What-Where-When Memory Test

Episodic memory is a complex memory system which allows recall and mental re-experience of previous episodes from one&#39;s own life. Real-life episodic ...

The (Spatial) Memory Game: Testing the Relationship Between Spatial Language, Object Knowledge, and Spatial Cognition

The Spatial Memory Game: Testing the Relationship Between Spatial Language, Object Knowledge, and Spatial Cognition

The memory game paradigm is a behavioral procedure to explore the relationship between language, spatial memory, and object knowledge. Using two different ...