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. 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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 ...

The overall goal of this behavioral assay is to test various aspects of learning and memory in mice. This method can help answer numerous questions in the area of rodent cognition, from the basics of developing a deeper understanding, for certain aspects of learning in memory and mice, as well as more complex issues, such as cognitive impairment, including disease models, such as Alzheimer's disease, to the exploration of potential cognitive enhancement. The main advantage of the ORT, is its simplicity, efficiency and flexibility and studying learning and memory in mice.Demonstrating this procedure will be Andrei Jeltyi, a lab technician from our lab. Begin by selecting mole sized objects that can be easily discriminated by the animal but have a similar degree of complexity in features, such as, texture, shape or brightness to minimize any potential induced object preference that may bias the results. Additionally, to further reduce induced preferences, ensure that mice are able to climb on both objects or neither object.The ability to climb on an object may increase interest in exploration. Then to minimize the stress of bright lighting, use defused low lighting with the center of the maze illuminated around 20 lucks. Acclimate mice to any new testing rooms for at least one hour prior to use each day and keep the temperature and humidity similar to regular housing conditions.Note, the mice should be handled one to two times a day for at least one minute, over the course of one to two weeks prior to testing. Lastly, for the main arena, use a square chamber made from white or black non-porous plastic contrasted to the color of the mouse. Begin by removing the mouse from its home cage and placing it in the middle of the open empty arena.Allow for free exploration of the arena for five minutes. Once the home cage is empty, save it for use as a holding cage the next day. At the end of five minutes, remove the mouse and place it in a holding cage.Do not return the mouse to its original home cage, or this may affect the behavior of the remaining mice to be tested. Then thoroughly clean the apparatus between mice, using 70%volume per volume ethanol. Next, begin the training phase or T1, by placing two identical objects in opposite quadrons of the arena.For example, one in the north west corner and one in the south east corner. Fasten any objects that are too light to the floor with removal mounting putty. Counter balance the use of each set of objects as well as the location of the novel object to each of the four corners of the arena.Be sure to know, which diagonal is used for which animal, so that the diagonal use on the training day is the shape as that used on the testing day for each mouse. 24 hours after habituation, remove the mouse from its home cage and place it in the center of the arena, equidistant from the two identical objects to begin training. Allow free exploration for a minimum of five minutes, if fusing a strain of mice that are known to have low locomotor or exploration activity, extend the trial to 10 minutes for all mice in the cohorts.At the end of training trial, remove the mouse and place it in the holding cage. Once the home cage is empty, save it for use as the holding cage on testing day. Be sure to clean the apparatus and objects between mice.Next, start the testing phase or T2, by placing the familiar object used during training and one novel object in opposite quadrons of the arena. Use the same locations as you use during training for each mouse. At the T1 to T2 interval of choosing, remove the mouse from its home cage and place it in the center arena, equidistant from the familiar object and the novel object.Allow free exploration for 10 minutes. Finally, at the end of the testing trial, remove the mouse and place it in the holding cage. For analysis, exploration is defined as the nose directed towards the object within two to three centimeters with active vibracy sweeping.However, the mouse sitting on the object, is not counted towards exploration time. For both T1 and T2, score the first five minutes and calculate the total exploration time for both objects for each session. If the mouse does not meet the minimum exploration time of 20 seconds for both objects, continue scoring past five minutes until total exploration exceeds 20 seconds.Extend T1 and T2 time to 10 minutes for strains of mice that have low exploration and do not meet this minimum criterion by five minutes as observed during pilot testing. Finally, if mice do not reach a 20 second minimum of exploration for both objects for either T1 or T2 at 10 minutes, exclude them from the analysis as it cannot be confirmed that the animal spent enough time exploring to learn and discriminate. The object recognition task is an efficient and flexible method for studying learning and memory in mice, in which phosphodiesterase 2 inhibitors have been shown to improve during this task.As compared to vehicle, administration of the PDE2 inhibitors bays 60-7550 or ND7001, significantly enhanced memory in a dose dependent manner, when given 30 minutes prior to training. When administered at different time points, relative to training and testing, the PDE2 inhibitor bay 60-7550 significantly enhanced his memory when given 30 minutes prior to training, as well as immediately after training and 30 minutes prior to recall. Once mastered, this technique can be completed in two to three days if it's performed properly.While attempting this procedure, it's important to remember to run pilot studies and carefully choose the T1 to T2 interval, as choosing a wrong interval may result in a misaffect. Following this procedure, other methods like the Barn's maze can be performed in order to answer additional questions like differences in learning, which cannot be fully addressed using the object recognition tests. After its development, this technique paved the way for researchers in the field of learning and memory to explore the basic biological mechanisms of memory, as well as explore potential mechanisms of various cognitive deficits in rodents.After watching this video, you should have a good understanding of how to set up and design an appropriate experiment using the object recognition test, including running pilot studies, choosing objects and selecting the appropriate T1 to T2 interval. Thanks for watching and good luck with your experiments.

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Research

JoVE Journal

Behavior

The Successive Alleys Test of Anxiety in Mice and Rats

The plus-maze was derived from the early work of Montgomery. He observed that rats tended to avoid the open arms of a maze, preferring the enclosed ones. Handley, Mithani and File et al. performed the first studies on the plus-maze design we use today, and in 1987 Lister published a design for use with mice. 
Time spent on, and entries into, the open arms are an index of anxiety; the lower these indices, the more anxious the mouse is. Alternatively, a mouse that spends most of its time in the closed arms is classed as anxious. 
One of the problems of the plus-maze is that, while time spent on, and entries into, the open arms is a fairly unambiguous measure of anxiety, time in the central area is more difficult to interpret, although time spent here has been classified as &#x201C;decision making&#x201D;. In many tests central area time is a considerable part of the total test time.
Shepherd et al. produced an ingenious design to eliminate the central area, which they called the &#x201C;zero maze&#x201D;. However, although used by several groups, it has never been as widely adopted as the plus-maze. 
In the present article I describe a modification of the plus-maze design that not only eliminates the central area but also incorporates elements from other anxiety tests, such as the light-dark box and emergence tests. It is a linear series of four alleys, each having increasing anxiogenic properties. It has given similar results to the plus-maze in general. Although it may not be more sensitive than the plus-maze (more data is needed before a firm conclusion can be reached on this point), it provides a useful confirmation of plus-maze results which would be useful when, for example, only a single example of a mutant mouse was available, as, for example, in ENU-based mutagenesis programs.

The plus-maze measures anxiety-like behaviour in rodents. There are two opposite closed and two opposite open arms; anxious rodents avoid the open arms. The central area is neither completely open nor closed, so time spent here is ambiguous and difficult to interpret. Here a modification of the plus-maze protocol eliminating this area is described.

The plus-maze was derived from the early work of Montgomery. He observed that rats tended to avoid the open arms of a maze, preferring the enclosed ones.  ...

The Double-H Maze: A Robust Behavioral Test for Learning and Memory in Rodents

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 behavioral flexibility and required memory duration, the number of goals and pathways, and also the overall task complexity. A confounding feature in many of these tasks is the lack of control over the strategy employed by the rodents to reach the goal, e.g., allocentric (declarative-like) or egocentric (procedural) based strategies. The double-H maze is a novel water-escape memory task that addresses this issue, by allowing the experimenter to direct the type of strategy learned during the training period. The double-H maze is a transparent device, which consists of a central alleyway with three arms protruding on both sides, along with an escape platform submerged at the extremity of one of these arms.
Rats can be trained using an allocentric strategy by alternating the start position in the maze in an unpredictable manner (see protocol 1; &#167;4.7), thus requiring them to learn the location of the platform based on the available allothetic cues. Alternatively, an egocentric learning strategy (protocol 2; &#167;4.8) can be employed by releasing the rats from the same position during each trial, until they learn the procedural pattern required to reach the goal. This task has been proven to allow for the formation of stable memory traces.
Memory can be probed following the training period in a misleading probe trial, in which the starting position for the rats alternates. Following an egocentric learning paradigm, rats typically resort to an allocentric-based strategy, but only when their initial view on the extra-maze cues differs markedly from their original position. This task is ideally suited to explore the effects of drugs/perturbations on allocentric/egocentric memory performance, as well as the interactions between these two memory systems.

The goal of this protocol is to investigate spatial cognition in rodents. The double-H water maze is a novel test, which is particularly useful to elucidate the different components of learning, consolidation and memory, as well as the interplay of memory systems.

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, 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.

 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 commonly split into lower-order and higher-order components, with lower order RRBs consisting of motor stereotypies and higher order RRBs consisting of perseverative and sequencing behaviors. Higher order RRBs are challenging to model in mice. Current assays for RRBs in mice focus primarily on the lower order components, making basic biomedical research into potential treatments or interventions for higher-order RRBs difficult. Here we describe a new assay, novel object exploration. This assay uses a basic open-field arena with four novel objects placed around the perimeter. The test mouse is allowed to freely explore the arena and the order in which the mouse investigates the novel objects is recorded. From these data, patterned sequences of exploration can be identified, as can the most preferred object for each mouse. The representative data shared here and past results using the novel object exploration assay illustrate that inbred mouse strains do demonstrate different behavior in this assay and that strains with elevated lower order RRBs also show elevated patterned behavior. As such, the novel object exploration assay appears to possess good face validity for higher order RRBs in humans and may be a valuable assay for future studies investigating novel therapeutics for ASD.

Higher order restricted, repetitive behaviors (RRBs) disrupt the lives of affected individuals. These behaviors are challenging to model in rodents, making basic biomedical research into potential treatments or interventions for RRBs difficult. Here we describe novel object exploration as a potential assay for higher order RRBs in mice.

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 four mountains within a computer-generated landscape is tested using a delayed match-to-sample paradigm. Allocentric spatial memory is assessed by altering the viewpoint, colors and textures between the initially presented and target images.
Allocentric spatial memory is a key function of the hippocampus, one of the earliest brain regions to be affected in Alzheimer&#39;s disease (AD) and impairment of hippocampal function predates the onset of dementia. It was hypothesized that performance on the 4MT would aid the diagnosis of predementia AD, which manifests clinically as Mild Cognitive Impairment (MCI).
The 4MT was applied to patients with MCI, stratified further based on cerebrospinal fluid (CSF) AD biomarker status (10 MCI biomarker positive, 9 MCI biomarker negative), and with mild AD dementia, as well as healthy controls. Comparator tests included tests of episodic memory and attention widely accepted as sensitive measures of early AD. Behavioral data were correlated with quantitative MRI measures of the hippocampus, precuneus and posterior cingulate gyrus.
4MT scores were significantly different between the two MCI groups (p = 0.001), with a test score of &#8804;8/15 associated with 100% sensitivity and 78% specificity for the classification of MCI with positive AD biomarkers, i.e., predementia AD. 4MT test scores correlated with hippocampal volume (r = 0.42) and cortical thickness of the precuneus (r = 0.55).
In conclusion, the 4MT is effective in identifying the early stages of AD. The short duration, easy application and scoring, and favorable psychometric properties of the 4MT fulfil the need for a simple but accurate diagnostic test for predementia AD.

The 4 Mountains Test: A Short Test of Spatial Memory with High Sensitivity for the Diagnosis of Pre-dementia Alzheimer&#039;s Disease

This article describes the 4 Mountains Test (4MT), a hippocampus-dependent test of working allocentric spatial memory. The hippocampus is affected early in Alzheimer's disease (AD) and this article outlines the 4MT methodology and results of patient testing, which demonstrates the value of the 4MT in the diagnosis of pre-dementia AD.

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 ...

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 memories are about events in their spatiotemporal context and are typically visuospatial, rather than verbal. Yet often, tests of episodic memory use verbal material to be recalled (word lists, stories). The Real-World What-Where-When memory test requires participants to hide a total of 16 different objects in 16 different locations over two temporal occasions, 2 h apart. Another two hours later, they are then asked to recall which objects (What) they had hidden in which locations (Where) and on which of the two occasions (When). In addition to counting the number of correctly recalled complete what-where-when combinations, this task can also be used to test real-world spatial memory and object memory. This task is sensitive to normal cognitive aging, and correlates well with performance on other episodic memory tasks, while at the same time providing more ecological validity and being cheap and easy to run.

A Real-world What-Where-When Memory Test

The Real-World What-Where-When memory test is a novel episodic memory test, in which participants need to recall which objects have been hidden in which locations on which of two distinct occasions. It is easy to run and is sensitive to normal cognitive aging.

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 memory game paradigm is a behavioral procedure to explore the relationship between language, spatial memory, and object knowledge. Using two different versions of the paradigm, spatial language use and memory for object location are tested under different, experimentally manipulated conditions. This allows us to tease apart proposed models explaining the influence of object knowledge on spatial language (e.g., spatial demonstratives), and spatial memory, as well as understanding the parameters that affect demonstrative choice and spatial memory more broadly. Key to the development of the method was the need to collect data on language use (e.g., spatial demonstratives: &#34;this/that&#34;) and spatial memory data under strictly controlled conditions, while retaining a degree of ecological validity. The language version (section 3.1) of the memory game tests how conditions affect language use. Participants refer verbally to objects placed at different locations (e.g., using spatial demonstratives: &#34;this/that red circle&#34;). Different parameters can be experimentally manipulated: the distance from the participant, the position of a conspecific, and for example whether the participant owns, knows, or sees the object while referring to it. The same parameters can be manipulated in the memory version of the memory game (section 3.2). This version tests the effects of the different conditions on object-location memory. Following object placement, participants get 10 seconds to memorize the object&#39;s location. After the object and location cues are removed, participants verbally direct the experimenter to move a stick to indicate where the object was. The difference between the memorized and the actual location shows the direction and strength of the memory error, allowing comparisons between the influences of the respective parameters.

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

We present a protocol to explore the relationship between spatial language production, spatial memory, and object knowledge. The procedure allows experimental manipulation of, and control over, conditions of object knowledge, language at instruction, and physical location, thus teasing apart cognitive and linguistic models describing interactions between these variables.

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

A Behavioral Assay for Investigating the Role of Spatial Memory During Instinctive Defense in Mice

Evolution has selected a repertoire of defensive behaviors that are essential for survival across all animal species. These behaviors are often stereotyped actions elicited in response to innately aversive sensory stimuli, but their success requires enough flexibility for adapting to different spatial environments, which can change rapidly. Here, we describe a behavioral assay to evaluate the influence of learned spatial knowledge on defensive behaviors in mice. We have adapted the widely used Barnes maze spatial memory assay to investigate how mice navigate to a shelter during escape responses to innately aversive sensory stimuli in a novel environment, and how they adapt to acute changes in the environment. This new assay is an ethological paradigm that does not require training and exploits the natural exploration patterns and navigation strategies in mice. We propose that the set of protocols described here are a powerful means of studying goal-directed behaviors and stimulus-triggered navigation, which should be of interest to both the fields of instinctive behaviors and spatial memory.

This protocol describes a behavioral assay, based on the Barnes maze test, to study how instinctive defensive actions are modified by the knowledge of spatial environment.

Evolution has selected a repertoire of defensive behaviors that are essential for survival across all animal species. These behaviors are often ...

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.

Ethologically relevant behavioral testing is a critical component of any study that uses mouse models to study the cognitive effects of various ...

Defining the Role Of Language in Infants' Object Categorization with Eye-tracking Paradigms

Assessing infant category learning is a challenging but vital aspect of studying infant cognition. By employing a familiarization-test paradigm, we straightforwardly measure infants&#8217; success in learning a novel category while relying only on their looking behavior. Moreover, the paradigm can directly measure the impact of different auditory signals on the infant categorization across a range of ages. For instance, we assessed how 2-year-olds learn categories in a variety of labeling environments: in our task, 2-year-olds successfully learned categories when all exemplars were labeled&#160;or the first two exemplars were labeled, but they failed to categorize when no exemplars were labeled or only the final two exemplars were labeled. To determine infants&#8217; success in such tasks, researchers can examine both the overall preference displayed by infants in each condition and infants&#8217; pattern of looking over the course of the test phase, using an eye-tracker to provide fine-grained time-course data. Thus, we present a powerful paradigm for identifying the role of language, or any auditory signal, in infants&#8217; object category learning.

Defining the Role Of Language in Infants&#039; Object Categorization with Eye-tracking Paradigms

Here we present a protocol for familiarization-test paradigms which provide a direct test of infant categorization and help to define the role of language in early category learning.

Assessing infant category learning is a challenging but vital aspect of studying infant cognition. By employing a familiarization-test paradigm, we ...