The authors thank Dr. Cora SW Lai from the School of Biomedical Sciences, the University of Hong Kong, for lending the elevated plus maze test, and the Department of Anesthesiology from the University of Hong Kong for lending the rotarod test apparatus.

All methods described here have been approved by the Committee on the Use of Live Animals in Teaching and Research (CULATR), the University of Hong Kong.
1. General protocol
NOTE: This section is based on Deacon19.
<ol>
	<li>Behavioral room setup
	<ol>
		<li>Get rid of unrelated stimulation/distraction, including direct bright light on the experimental apparatus, odor, noise, and other irrelevant animals, in the behavioral room (which should be about 10 m2 with adjustable lighting and, preferably, have an anteroom). 
		NOTE: Since the mouse is a nocturnal animal, lighting under 15 lux in the open-field test, novel object recognition test, and social interaction test could minimize the interference/stress from the light and help the mouse to focus on the test.</li>
		<li>Set the camera for video recording at least 1.5 m above the floor, ensuring that it is out of the sight of the testing mouse.</li>
	</ol>
	</li>
	<li>Housing and habituation
	<ol>
		<li>Group-house the mice in an animal unit under observation (e.g., group-housing no more than four adult mice). 
		NOTE: Here, 3-month-old male C57BL/6N mice were used and housed in a 1144B cage. Rule out sick, injured, or severely stressed mice. Experiences of starvation, thirst, or being bullied may affect the performance of the mouse.</li>
		<li>Arrange for the same animal handler to conduct all the behavioral tests, to diminish variability. Perform the transportation, handling, and experiment during the light cycle from 7:00 a.m. to 7:00 p.m. If possible, arrange all the other handling, such as the administration of any drug/toxin (e.g., intranasal instillation of silica nanoparticles) or cage-cleaning, after the test.</li>
		<li>Relabel the cages with random numbers to blind the experimenter before each experiment. Habituate the mice to the experimental environment in the behavioral room for 15 to 30 min in their home cages. Keep the home cages in the behavioral room during the entire experiment.</li>
		<li>Before starting the experiment, put a nonexperimental C57BL/6N mouse in the apparatus so that the experimental condition for the first mouse is the same as the rest. Then, clean the apparatus as follows: remove the urine and feces with a clean paper towel, clean the experimental device with tap water, and then, cover the odor left by the mouse by wiping the apparatus with a paper towel lightly sprayed with 70% ethanol.</li>
		<li>To minimize the distraction caused by the experimenter, ask the experimenter to leave the behavioral room during video recording, or stay behind the curtain during the Morris water maze test.</li>
	</ol>
	</li>
	<li>Behavioral test arrangement
	<ol>
		<li>Arrange the behavioral test in the order as shown in Figure 1A. Plan to perform the tests spaced out by 24 h, except for the elevated plus maze test. 
		NOTE: Since the entire procedure requires up to 2 weeks, simplify the battery by choosing among similar tests before application in an acute or short-term study. 
		CAUTION: The 2 day open-field test ensures proper habituation for the novel object recognition test. Test the forced swimming test last as it may cause stress in C57BL/6N mice.</li>
	</ol>
	</li>
</ol>
2. BehavioralTest Protocol
<ol>
	<li>Open-field test8,18,19,20
	<ol>
		<li>Perform the open-field test in a 60 cm (length [L]) x 60 cm (width [W]) x 40 cm (height [H]) nontransparent white plastic arena. 
		NOTE: Using multiple arenas can increase the throughput of the test.</li>
		<li>Start the camera recording and gently put the mouse next to the middle of a wall of the arena, facing that same wall. Record the behavior of the mouse for 10 min before returning it to the home cage. Clean the apparatus as described in step 1.2.4. 
		NOTE: The start point in the open-field test can also be the center of the arena. Be consistent among the mice.</li>
		<li>Repeat until all the mice finish the protocol. Counterbalance the testing order between groups.</li>
		<li>Perform the data analysis as follows.
		<ol>
			<li>Divide the arena into four squares by four squares (imaginary grid) on the computer screen. To assess locomotor function, count the number of lines crossed by the mouse in the arena19. 
			NOTE: The definition of &#8220;crossing a line&#8221; is when both hind limbs cross it. This definition also applies to the elevated plus maze test.</li>
			<li>Measure the time spent in the central area as an indicator of anxiety. The central area is the four squares in the center of the arena. 
			NOTE: Additional parameters, such as rearing (both front paws off the ground, with front paws against a wall or standing), latency to the first rear, and grooming and freezing indicate the emotionality of the mouse.</li>
			<li>Alternatively, use a tracking software, as described in detail by Seibenhener and Wooten8, to measure the distance traveled, the speed, and the time spent in the center area.</li>
		</ol>
		</li>
	</ol>
	</li>
	<li>Accelerating rotarod test18
	<ol>
		<li>Perform the 3 day training of the accelerating rotarod test before any treatment of drugs or toxin/modeling/onset of disease and test the motor function for 1 day, as planned in Figure 1A. 
		NOTE: The mouse receives three trials per day during the training and testing. Each trial starts with the rotation of the rod and ends with the drop of the mouse.</li>
		<li>Place the rotarod apparatus on the bench in the behavioral room. Avoid direct lighting to the equipment. Program the equipment as starting from 4 rpm and accelerating to 40 rpm within 5 min.</li>
		<li>In each trial, put the mouse on the static rod, facing the wall of the machine. Start the device when the mouse is settled. Stop the device once the mouse drops and record the time the mouse spent on the rod. Immediately repeat for another two trials before returning the mouse back to the home cage.</li>
		<li>Repeat the procedure on the other mice.</li>
		<li>Measure the average time spent on the rod of three trials during testing to estimate motor function. 
		NOTE: The average time on the rod during the third day of training is the baseline of motor function.</li>
	</ol>
	</li>
	<li>Social interaction test18
	<ol>
		<li>For the social interaction test, use an open-field arena with two identical transparent chambers (8 cm [L] x 6 cm [W] x 12 cm [H]) with holes on the surface&#8212;and a novel mouse (helper) which is a same-sex juvenile conspecific that has had no previous contact with the subject mouse. Figure 1B shows the scheme of the procedure. Clean the arena and the chambers as described in step 1.2.4. 
		NOTE: The novel mouse cannot be a littermate or cage mate of the subject mouse. It is group-housed and healthy. Habituate the novel mouse to the behavioral room for 15 to 30 min as described in step 1.2.3.</li>
		<li>Separately place the two chambers in the middle of two opposite walls of the arena. Introduce the subject mouse into the arena as described in step 2.1.2 and shown in Figure 1B, for a 3 min exploration. Return the subject mouse to the home cage and remove any urine or feces in the arena.</li>
		<li>Put the helper in one of the chambers. Reintroduce the subject mouse to the arena and record for 3 min. Afterward, return both mice to each of their own home cages. Repeat the procedures with other subject mice as described above. 
		NOTE: Counterbalance the side of the helper or randomly assign it within the group.</li>
		<li>From the video, estimate the parameter describing the social interaction activity of the mice as thelper/tempty, which means the ratio of time interacting with the helper chamber (thelper) and the empty one (tempty), or use the recognition index thelper/(thelper + tempty). 
		NOTE: An interaction between the subject mouse and the chamber is defined as when the mouse&#8217;s nose is within 2 cm of the chamber and pointing toward it.</li>
	</ol>
	</li>
	<li>Elevated plus maze test10
	<ol>
		<li>Conduct the elevated plus maze test on the same day after all mice are tested in the open-field test. Clean the apparatus as described in step 1.2.4. 
		NOTE: The configuration of the elevated plus maze is a &#8220;+&#8221;-shape. It has two open arms (30 cm x 5 cm x 0.5 cm) across from each other and perpendicular to two closed arms (30 x 5 x 16 cm) with a center platform (5 cm x 5 cm x 0.5 cm). The maze is elevated 40 cm from the ground.</li>
		<li>Place the mouse at the junction of the open and closed arms, facing the open arm that is opposite to the experimenter (Figure 1C). Record the behavior for 5 min before returning the mouse to the home cage. Repeat till all mice are tested. 
		NOTE: Entering the maze with its face to the open arm could increase the mouse&#8217;s exploration of the open arm.</li>
		<li>Measure the time the mouse spent in the open arms (topen) and in the closed arms (tclose) based on the video: topen/tclose indicates the level of anxiety.</li>
	</ol>
	</li>
	<li>Forced swim test11
	<ol>
		<li>The apparatus of the forced swim test is a cylindrical tank that is 30 cm high and 20 cm in diameter. Fill the tank up to 15 cm high with tap water at room temperature (23 - 25 &#176;C). 
		NOTE: Use fresh water for each mouse.</li>
		<li>Start the video recording and gently put the mouse in the water, in the center of the apparatus. Record the video for 6 min before putting the mouse back in its home cage under infrared light. 
		NOTE: Do not disturb the mouse by drowning it or twisting its tail.</li>
		<li>Measure the immobility time in the last 5 min of the recorded video. Mobility means any movements other than those required to balance the body and to keep the head above the water.</li>
	</ol>
	</li>
	<li>Novel object recognition test17,18
	<ol>
		<li>Set up the novel object recognition test to include 2 days of habituation, 1 day of familiarization, and 1 day of testing (Figure 1D); each session is 10 min per mouse, and the intersession interval is 24 h. 
		NOTE: The habituation using the open-field test is performed as described in section 2.1. The mouse interacts with two identical objects (old objects) in familiarization. In the test, the mouse interacts with one of the old objects and a new object, both placed in the same place as the objects in familiarization. The apparatus of the novel object recognition test includes an open-field arena and two sets of objects. Each set contains two identical objects (objects A and A and objects B and B). Objects A and B are similar in size but different in texture (glass/plastic/paper), shape (round/cubic), and color (bright/dark). The objects should be odor-free and big enough for the mouse to explore within 10 min. The appropriate size for adult C57BL/6N mice is 8 cm tall and 5 cm wide/in diameter.</li>
		<li>Mark the positions of the two objects in familiarization and test, which are 5 cm away from the side and 7 cm away from the top of the arena. 
		NOTE: Mark the position on the evening before the familiarization to avoid the smell of the marker.</li>
		<li>In the familiarization, the mouse interacts with one set of identical objects. Clean the arena and objects as described in step 1.2.4 before placing the mouse in the arena, facing the middle of the wall as shown in Figure 1D. Record for 10 min before returning the mouse to the home cage. Repeat until all the mice are finished and return all the cages to the animal unit. 
		NOTE: Counterbalance the objects used in familiarization within the group to diminish bias (e.g., mice No. 1 and 2 explore objects A and A, and mice No. 3 and 4 explore objects B and B; in this way, the novel object is object B for mice No. 1 and 2 and object A for mice No. 3 and 4).</li>
		<li>Perform the test 24 h after the familiarization. Use the same procedure as for the familiarization, except replace one of the objects with one from another set (Figure 1D). Repeat until all the mice have performed the test and, afterward, return all the cages to the animal unit. 
		NOTE: Counterbalance the side of the new object within the group to diminish bias (e.g., introduce mice No. 1 and 3 to objects A and B, and mice No. 2 and 4 to objects B and A). In this way, the novel object shows at the right side for mice No. 1 and 4, and at the left side for mice No. 2 and 3. Here, the left side is the left side of the experimenter when facing the arena.</li>
		<li>Measure the time that each mouse interacts with the new object (tnew) and the old object (told) separately, from the video footage in the test phase. An interaction between the animal and the object is described in the note following step 2.3.4. Calculate the memory of the mouse as the preference to the novel object = tnew/told; or tnew/(tnew + told). 
		NOTE: tnew/told equals to 1 or tnew/(tnew + told) equals to 0.5 means the mouse has no preference for the novel object (i.e., memory impairment). The time interacting with objects in the familiarization can serve as a control of the experiment. The total time indicates the exploration activity of the mouse, and tleft/tright suggests spatial bias.</li>
	</ol>
	</li>
	<li>Morris water maze test16
	<ol>
		<li>Set up the apparatus as follows.
		<ol>
			<li>Put the water maze, a circular pool (of 120 cm in diameter and 60 cm deep), in the center of a behavioral room, and mark the position of the maze to ensure the position remains the same during the entire experiment.</li>
			<li>Divide the maze into four equal imaginary quadrants. Hang the visual cues (e.g., circle, square, triangle, and pentagon) in the center of each quadrant, 130 cm above the floor and 53 cm away from the wall of the maze. 
			NOTE: The maze and the cue must stay in the same position during the entire test, so the mouse can form accurate spatial memory.</li>
			<li>Place a platform 25 cm away from the wall, in the center of the fourth quadrant, and mark the position. The platform for the mouse is 10 cm in diameter. 
			NOTE: The position and diameter of the platform determine the difficulty of the task. The nearer it is to the wall of the maze, or the bigger the platform, the easier the task.</li>
			<li>Fill the water maze with water (with a temperature of 23 to 25 &#176;C, colored into white and made opaque by milk powder/food whitening powder) until the water level is 1 cm higher than the platform. Bring the mice into the behavioral room for 15 to 30 min of habituation as shown in step 1.2.3 and turn on the infrared light above the cages, which will be used to dry the mice. 
			NOTE: Cover the top of the platform with white cloth and net so that the mouse can easily climb onto it. Make sure there is no direct lighting above the water.</li>
		</ol>
		</li>
		<li>Conduct the training phase as follows.
		<ol>
			<li>The training phase takes 5 days, four trials per day. Semi-randomly arrange the starting points on each day as demonstrated in literature16. This effort prevents the mouse from establishing associative memory, which is the most common way to &#8220;cheat&#8221; in the test.</li>
			<li>At the beginning of each trial, start the video recording and gently put the mouse into the maze. 
			NOTE: Do not drop the mouse into the tank or twist its tail, which may cause extra stress and disorientation.</li>
			<li>Ask the experimenter to stay out of sight of the mouse and to return to take the mouse back to its home cage when any of the following happens: (i) the mouse cannot locate the platform within 60 s; (ii) the mouse finds the platform within 60 s and stays on it for 10 s. In circumstance (i), ask the experimenter to place the mouse on the platform and let it stay there for 10 s. 
			NOTE: Point (ii) means the mouse successfully located the platform.</li>
			<li>Stop the video and put the mouse back in the home cage under infrared light. 
			NOTE: Maintaining the mice&#8217;s body temperature is critical for their performance because hypothermia stresses mice and may affect the following tests.</li>
			<li>Repeat the procedure with another mouse.</li>
			<li>Based on the video, record the escape latency, which is the duration of the period the mouse spends in the maze, from entering the maze till the moment it successfully locates the platform. If the mouse cannot find the platform or stays there for less than 10 s in 60 s, the escape latency counts as 60 s. Plot the learning curve against the training days with the average escape latency per day. 
			NOTE: The escape latency does not include the 10 s spend on the platform.</li>
		</ol>
		</li>
		<li>Perform the probe phase as follows.
		<ol>
			<li>On the sixth day of the Morris water maze test, set up the apparatus as described in step 2.7.1., take a picture of the maze to record the position of the platform, and then remove the platform from the tank.</li>
			<li>Start the video recording, and gently put the mouse into the maze in the quadrant diagonally opposite to the target quadrant.</li>
			<li>Ask the experimenter to stay out of sight of the mouse during the 1 min video recording. Afterward, have the experimenter take the mouse out of the maze and put it back in the home cage.</li>
			<li>Use the image taken at step 2.7.3.1. as a reference to measure the duration of any platform crossing. Time the duration the mouse stays in the target quadrant (ttarget), according to the video. The total time is ttotal. Measure the preference to the target quadrant as ttarget /ttotal.</li>
		</ol>
		</li>
	</ol>
	</li>
</ol>

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V., 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=Behavioural+battery+testing:+evaluation+and+behavioural+outcomes+in+8+inbred+mouse+strains.">Behavioural battery testing: evaluation and behavioural outcomes in 8 inbred mouse strains.</a> Physiology &amp; Behavior. 99 (3), 301-316 (2010).</li><li>Powell, T. R., Fernandes, C., Schalkwyk, L. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Depression-Related+Behavioral+Tests.">Depression-Related Behavioral Tests.</a> Current Protocols in Mouse Biology. 2 (2), 119-127 (2012).</li><li>Paylor, R., Spencer, C. M., Yuva-Paylor, L. A., Pieke-Dahl, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+use+of+behavioral+test+batteries,+II:+effect+of+test+interval.">The use of behavioral test batteries, II: effect of test interval.</a> Physiology &amp; Behavior. 87 (1), 95-102 (2006).</li><li>Lee, K. M., Coehlo, M., McGregor, H. A., Waltermire, R. S., Szumlinski, K. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Binge+alcohol+drinking+elicits+persistent+negative+affect+in+mice.">Binge alcohol drinking elicits persistent negative affect in mice.</a> Behavioural Brain Research. 291, 385-398 (2015).</li><li>Shiotsuki, H., 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=A+rotarod+test+for+evaluation+of+motor+skill+learning.">A rotarod test for evaluation of motor skill learning.</a> Journal of Neuroscience Methods. 189 (2), 180-185 (2010).</li></ol>

The authors have nothing to disclose.

Behavioral analysis of mice is critical for neurodegeneration research. While cognitive function is often the most susceptible domain of behavior affected in neurodegenerative diseases, mood dysfunction, such as depression and anxiety, is often comorbid. Moreover, motor function often affects the interpretation of the results in some tests, such as the novel object recognition test, the elevated plus maze test, and the social interaction test. Based on these thoughts, a comprehensive behavioral test battery is required f...

Neurodegenerative diseases featured devastating behavioral symptoms, including cognitive impairment, mood dysfunctions such as anxiety and depression, or motor dysfunction1. The pathogenesis of various kinds of neurodegenerative diseases is unclear2. Accumulative studies indicate that genetic and environmental factors might both contribute to the pathogenesis of neurodegenerative diseases. Identifying the risk factor of neurodegeneration requires behavioral analysis. Although each type of neurodegenerative disease has its signature behavioral symptom (e.g.,&#160;Alzheimer&#39;s disease [AD] is featured with cognitive impairment and Parkinson&#39;s disease [PD] with motor dysfunction). With the progression of the disease, the patients manifest comorbidity of different behavioral abnormalities3. For example, AD patients show symptoms of mood dysfunction in the advanced stage4,5. PD patients may progress into PD-related dementia and develop cognitive impairment6. Based on these features, the behavioral analysis in neurodegeneration models is usually comprehensive and repeated.
To achieve this goal, a battery which contains classical and widely used behavioral tests with excellent validity was designed for behavioral analyses in motor, mood, and cognition. The motor function can be tested by the open-field test7,8 and the accelerating rotarod test. Mood dysfunction, including social dysfunction, depression, and anxiety, are most commonly seen in neurodegenerative diseases5. Hence, this battery includes a social interaction test for sociability9, the elevated plus maze test for anxiety10, and the forced swim test for depression11. Cognitive impairment is one of the most characteristic symptoms in neurodegenerative diseases such as AD and frontotemporal lobar dementia12. Cognitive domains, including short-term memory and episodic memory, are susceptible to neurodegeneration13,14,15. Therefore, the Morris water maze test for spatial learning and memory16 and the novel object recognition test for short-term memory17 are included in the battery. These tests are compatible with each other. The order of the tests was designed to maximize habituation and to minimize interference, to further increase the compatibility within the battery. Since each function is tested by at least two independent tests that are different in principle and method, the results of each test can be further validated. Moreover, the protocols of some tests are highlighted for repeated testing, facilitating the longitudinal study of the development of neurodegenerative diseases. Therefore, this behavior test battery studies different subdomains of behavioral changes seen in various stages of neurodegeneration while costing a minimal number of animals. This battery has been used in a longitudinal study which evaluated the behavioral changes in young adult (3-month-old) male C57BL/6N mice after respiratory exposure to silica nanoparticles, an occupational hazard that is a potential risk factor of neurodegeneration18. However, other strains or models, such as aged mice and genetically manipulated mice, may behave differently than young C57BL/6N mice. Therefore, caution may be required when using this battery in these mice.

<table>
	<tbody>
		<tr>
			<td>chambers in social interaction test</td>
			<td>home made</td>
			<td></td>
			<td>(8 cm (L) x 6 cm (W) x 12 cm (H)), transparant with holes, plastic</td>
		</tr>
		<tr>
			<td>cylindrical tanks used in forced swimming test</td>
			<td>home made</td>
			<td></td>
			<td>30 cm height, 20 cm diameters, glass</td>
		</tr>
		<tr>
			<td>elevated plus maze</td>
			<td>home made</td>
			<td></td>
			<td>open arms (30 x 5 x 0.5 cm) ,closed arms (30 x 5 x 16 cm), center platform (5 x 5 x 0.5 cm), 40 cm tall. Plastic, nontransparant</td>
		</tr>
		<tr>
			<td>IITC Roto-Rod Apparatus</td>
			<td>IITC life science Inc.</td>
			<td>755, series 8</td>
			<td></td>
		</tr>
		<tr>
			<td>open field arena</td>
			<td>home made</td>
			<td></td>
			<td>60 cm (L) x 60 cm (W) x 40 cm (H), plastic, nontransparant</td>
		</tr>
		<tr>
			<td>water maze</td>
			<td>home made</td>
			<td></td>
			<td>120 cm in diameter, 60 cm deep, steel</td>
		</tr>
	</tbody>
</table>

a behavioral test battery for the repeated assessment of motor skills, mood, and cognition in mice

Pharmacological and toxicological studies in neurodegeneration require comprehensive behavioral analysis in mice because motor dysfunctions and dysfunctions in mood and cognition are common and often shared symptoms in neurodegenerative diseases. Shown here is a behavioral test battery for motor, mood, and cognition, which can be repeatedly tested in a longitudinal study. This battery assesses the overall behavioral phenotype in mice by examining each domain of behavior with at least two independent well-accepted tests (i.e., open-field test and rotarod test for motor function, social interaction test, elevated plus maze test, and forced swim test for emotional function, and Morris water maze test and novel object recognition test for cognitive function). Therefore, this sensitive and comprehensive test battery is a powerful tool for the study of behavioral alternation in neurodegeneration.

This behavioral test battery was designed for the comprehensive and valid behavioral analysis of motor, mood, and cognition, which are commonly affected in neurodegeneration5. We have applied this battery to study the behavioral changes in young adult C57BL/6N mice after respiratory exposure to silica nanoparticles for 1 month and 2 months18. The results revealed that C57BL/6N mice exposed to silica nanoparticles showed various behavioral ch...

Watch this Scientific Journal Video about A Behavioral Test Battery for the Repeated Assessment of Motor Skills, Mood, and Cognition in Mice at JoVE.com

A Behavioral Test Battery for the Repeated Assessment of Motor Skills, Mood, and Cognition in Mice

A comprehensive behavioral test battery of motor skills, mood&#8212;including social interaction, depression, and anxiety&#8212;and cognition is designed for the repeated assessment of neurodegeneration-related behavioral changes in mice.

Pharmacological and toxicological studies in neurodegeneration require comprehensive behavioral analysis in mice because motor dysfunctions and ...

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Behavior

18.8K Views.  Children's Hospital of Nanjing Medical University. A comprehensive behavioral test battery of motor skills, mood—including social interaction, depression, and anxiety—and cognition is designed for the repeated assessment of neurodegeneration-related behavioral changes in mice.

Video: A Behavioral Test Battery for the Repeated Assessment of Motor Skills, Mood, and Cognition in Mice

The Resident-intruder Paradigm: A Standardized Test for Aggression, Violence and Social Stress

This video publication explains in detail the experimental protocol of the resident-intruder paradigm in rats. This test is a standardized method to measure offensive aggression and defensive behavior in a semi natural setting. The most important behavioral elements performed by the resident and the intruder are demonstrated in the video and illustrated using artistic drawings. The use of the resident intruder paradigm for acute and chronic social stress experiments is explained as well. Finally, some brief tests and criteria are presented to distinguish aggression from its more violent and pathological forms.

This video shows the resident-intruder paradigm in rats. This test is a standardized method to measure offensive aggression, defensive behavior and violence in a semi-natural setting. The use of the paradigm for social stress experiments is explained as well.

This video publication explains in detail the experimental protocol of the resident-intruder paradigm in rats. This test is a standardized method to ...

The Crossmodal Congruency Task as a Means to Obtain an Objective Behavioral Measure in the Rubber Hand Illusion Paradigm

The rubber hand illusion (RHI) is a popular experimental paradigm. Participants view touch on an artificial rubber hand while the participants' own hidden hand is touched. If the viewed and felt touches are given at the same time then this is sufficient to induce the compelling experience that the rubber hand is one's own hand. The RHI can be used to investigate exactly how the brain constructs distinct body representations for one's own body. Such representations are crucial for successful interactions with the external world. To obtain a subjective measure of the RHI, researchers typically ask participants to rate statements such as "I felt as if the rubber hand were my hand". Here we demonstrate how the crossmodal congruency task can be used to obtain an objective behavioral measure within this paradigm.
The variant of the crossmodal congruency task we employ involves the presentation of tactile targets and visual distractors. Targets and distractors are spatially congruent (i.e. same finger) on some trials and incongruent (i.e. different finger) on others. The difference in performance between incongruent and congruent trials - the crossmodal congruency effect (CCE) - indexes multisensory interactions. Importantly, the CCE is modulated both by viewing a hand as well as the synchrony of viewed and felt touch which are both crucial factors for the RHI.
The use of the crossmodal congruency task within the RHI paradigm has several advantages. It is a simple behavioral measure which can be repeated many times and which can be obtained during the illusion while participants view the artificial hand. Furthermore, this measure is not susceptible to observer and experimenter biases. The combination of the RHI paradigm with the crossmodal congruency task allows in particular for the investigation of multisensory processes which are critical for modulations of body representations as in the RHI.

We demonstrate how an objective measure can be employed in the widely employed rubber hand illusion paradigm. This measure is obtained by modifying the well-established crossmodal congruency task. This task allows the investigation of multisensory processes which are critical for modulations of body representations as in the rubber hand illusion.

The rubber hand illusion (RHI) is a popular experimental paradigm. Participants view touch on an artificial rubber hand while the participants' own hidden ...

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

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

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

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

Compensatory Limb Use and Behavioral Assessment of Motor Skill Learning Following Sensorimotor Cortex Injury in a Mouse Model of Ischemic Stroke

Mouse models have become increasingly popular in the field of behavioral neuroscience, and specifically in studies of experimental stroke. As models advance, it is important to develop sensitive behavioral measures specific to the mouse. The present protocol describes a skilled motor task for use in mouse models of stroke. The Pasta Matrix Reaching Task functions as a versatile and sensitive behavioral assay that permits experimenters to collect accurate outcome data and manipulate limb use to mimic human clinical phenomena including compensatory strategies (i.e., learned non-use) and focused rehabilitative training. When combined with neuroanatomical tools, this task also permits researchers to explore the mechanisms that support behavioral recovery of function (or lack thereof) following stroke. The task is both simple and affordable to set up and conduct, offering a variety of training and testing options for numerous research questions concerning functional outcome following injury. Though the task has been applied to mouse models of stroke, it may also be beneficial in studies of functional outcome in other upper extremity injury models.

Mouse models have become increasingly popular in studies of behavioral neuroscience. As models advance, it is important to develop sensitive behavioral measures specific to the mouse. This protocol describes the Pasta Matrix Reaching Task, which is a skilled motor task for use in mouse models of stroke.

Mouse models have become increasingly popular in the field of behavioral neuroscience, and specifically in studies of experimental stroke. As models ...

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.

This protocol describes the modified hole board (mHB), which combines features from a traditional hole board and open field and is designed to measure ...

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 ...

Assessment of Social Cognition in Non-human Primates Using a Network of Computerized Automated Learning Device (ALDM) Test Systems

Fagot &#38; Paleressompoulle1 and Fagot &#38; Bonte2 have published an automated learning device (ALDM) for the study of cognitive abilities of monkeys maintained in semi-free ranging conditions. Data accumulated during the last five years have consistently demonstrated the efficiency of this protocol to investigate individual/physical cognition in monkeys, and have further shown that this procedure reduces stress level during animal testing3. This paper demonstrates that networks of ALDM can also be used to investigate different facets of social cognition and in-group expressed behaviors in monkeys, and describes three illustrative protocols developed for that purpose. The first study demonstrates how ethological assessments of social behavior and computerized assessments of cognitive performance could be integrated to investigate the effects of socially exhibited moods on the cognitive performance of individuals. The second study shows that batteries of ALDM running in parallel can provide unique information on the influence of the presence of others on task performance. Finally, the last study shows that networks of ALDM test units can also be used to study issues related to social transmission and cultural evolution. Combined together, these three studies demonstrate clearly that ALDM testing is a highly promising experimental tool for bridging the gap in the animal literature between research on individual cognition and research on social cognition.

Assessment of Social Cognition in Non-human Primates Using a Network of Computerized Automated Learning Device ALDM Test Systems

Fagot &#38; Paleressompoulle1 have published an automated learning device (ALDM) aimed at testing individual cognitive abilities in semi-free ranging monkeys. The main goal of our protocol is to use a network of ALDM test units to study social cognition in non-human primates.

Fagot &#38; Paleressompoulle1 and Fagot &#38; Bonte2 have published an automated learning device (ALDM) for the study of cognitive abilities of monkeys ...

Assessment of Cocaine-induced Behavioral Sensitization and Conditioned Place Preference in Mice

It is thought that rewarding experiences with drugs create strong contextual associations and encourage repeated intake. In turn, repeated exposures to drugs of abuse make lasting alterations in the brain function of vulnerable individuals, and these persistent alterations likely serve to maintain the maladaptive drug seeking and taking behaviors characteristic of addiction/dependence2. In rodents, reward experience and contextual associations are frequently measured using the conditioned place preference assay, or CPP, wherein preference for a previously drug-paired context is measured. Behavioral sensitization, on the other hand, is an increase in a drug-induced behavior that develops progressively over repeated exposures. Since sensitized behaviors can often be measured after several months of drug abstinence, depending on the dose and length of initial exposure, they are considered observable correlates of lasting drug-induced plasticity. Researchers have found these assays useful in determining the neurobiological substrates mediating aspects of addiction as well as assessing the potential of different interventions in disrupting these behaviors. This manuscript describes basic, effective protocols for mouse CPP and locomotor behavioral sensitization to cocaine.

This protocol is intended to enable researchers to conduct experiments designed to test these aspects of addiction using the conditioned place preference and locomotor behavioral sensitization assays.

It is thought that rewarding experiences with drugs create strong contextual associations and encourage repeated intake. In turn, repeated exposures to ...

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 describes a rodent version of the PVT&#8212;termed the &#34;rPVT&#34;&#8212;that measures similar aspects of attention (i.e., performance accuracy, motor speed, premature responding, and lapses in attention). Data are presented that demonstrate both the short- and long-term usefulness of the rPVT when employed with laboratory rats. Rats easily learn the rPVT, and learning to perform the basic procedure takes less than two weeks of training. Once acquired, rat performances in the rPVT show a high degree of similarity to these same performance measures in the human PVT, including similarities in, lapses in attention, reaction times, vigilance decrements across session time (i.e., the human &#34;time-on-task&#34; effects), and the response-stimulus interval (RSI) effect described for humans. Thus the rPVT can be an extremely valuable tool for assessing the effects of a wide range of variables on sustained attention quite similar to human PVT performances, and thus can be useful for developing novel treatments for neurobehavioral dysfunctions.

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

A rat version of the human Psychomotor Vigilance Test (PVT) is described that measures aspects of attention similar to those measured with the human PVT, including aspects of human vigilance such as performance accuracy, motor speed, premature responding, and lapses in attention.

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

Assessing Working Memory in Children: The Comprehensive Assessment Battery for Children &#8211; Working Memory (CABC-WM)

The Comprehensive Assessment Battery for Children - Working Memory (CABC-WM) is a computer-based battery designed to assess different components of working memory in young school-age children. Working memory deficits have been identified in children with language-based learning disabilities, including dyslexia1,2 and language impairment3,4, but it is not clear whether these children exhibit deficits in subcomponents of working memory, such as visuospatial or phonological working memory. The CABC-WM is administered on a desktop computer with a touchscreen interface and was specifically developed to be engaging and motivating for children. Although the long-term goal of the CABC-WM is to provide individualized working memory profiles in children, the present study focuses on the initial success and utility of the CABC-WM for measuring central executive, visuospatial, phonological loop, and binding constructs in children with typical development. Immediate next steps are to administer the CABC-WM to children with specific language impairment, dyslexia, and comorbid specific language impairment and dyslexia.

Assessing Working Memory in Children: The Comprehensive Assessment Battery for Children &amp;#8211; Working Memory (CABC-WM)

Working memory predicts a significant amount of variance for a variety of cognitive tasks, including speaking, reading, and writing. However, few tools are available to assess working memory in children. We present an innovative, computer-based battery that comprehensively assesses different components of working memory in school-age children.

The Comprehensive Assessment Battery for Children - Working Memory (CABC-WM) is a computer-based battery designed to assess different components of ...