Name: a behavioral test battery for the repeated assessment of motor skills, mood, and cognition in mice
Uploaded: 2019-03-02T13:00:00
Description: 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

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, i...

<ol>
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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 ...

Neurodegenerative disease often manifests with devastating behavior symptoms, such as cognitive impairment, mood dysfunctions, or even motor dysfunctions. This study allows a behavioral phenotypes to be assessed accurately with a high level of repeatability. We design a comprehensive battery that can be repeatedly utilized in behavior analysis in mice.This test asses motor functions, social interaction, emotional function, and cognitive function. By organizing the order and the interval of tests in this battery, we increased the testing efficiency and decreased the interference among tests and the stress on mice from frequent handling. To begin, arrange the behavioral test in the order described in the text protocol.Perform the open field test in a non-transparent, white, plastic arena. Start the camera recording, and gently put the mouse next to the middle of a wall of the arena, facing the wall. Record the mouse's behavior for 10 minutes, and return it to its home cage.Next, divide the arena into four by four squares on the computer screen. Count the number of times the mouse crosses the gridlines in the arena to assess locomotor function. To gauge anxiety, measure the amount of time that the mouse spends in the central area of the arena.Conduct the elevated plus maze test on the same day. Once all mice finish the open field test, place the mouse at the junction of the open and closed arms. Record the mouse's behavior for five minutes before returning the mouse to its home cage.After two consecutive days of open field test, set up the novel object recognition test according to the text protocol. Start the camera, and place the mouse in the arena, facing the middle of the wall, to familiarize the mouse. Record the mouse's behavior for 10 minutes.Then return the mouse to its home cage. 24 hours after the familiarization, repeat the test with one novel object. Measure the time that the mouse interacts with a new object and a familiar object separately.Then calculate the mouse's memory as the preference to the novel object. For the social interaction test, use an open field arena with two transparent chambers with holes on the surface. Place the subject mouse in the arena for a three-minute objects exploration period to habituate the mouse to the chambers.Then return the subject mouse to its home cage and clean the arena. Put a novel mouse into one of the chambers. Then reintroduce the subject mouse to the arena and record for three minutes.While reviewing the video, gauge social interaction by measuring the ratio of time the subject mouse spends interacting with the novel mouse chamber or the empty chamber. Next perform the accelerating rotarod test. In each trial, place the mouse on the static rod facing the wall opposite to the machine.Habituate the mice to the rotarod by training it for three days in the naive mice before the onset of possible symptoms, so that the mouse equally learns the scale. Stop the device when the mouse drops, and record the time the mouse spent on the rod. Repeat this process two more times before returning the mouse to its home cage.Next put the water maze in the center of a behavioral room, and mark the position of the maze to ensure the position of the water maze remains constant. Divide the maze into four imaginary quadrants. Hang visual cues in the center of each quadrant.Then place a platform in the center of the fourth quadrant and mark its location. Fill the maze with water, colored white with milk powder, until the water level is one centimeter higher than the platform. At the beginning of each training trial, start the video recording, and gently put the mouse in the maze.Stay out of sight of the mouse. Return the mouse to its home cage when the mouse cannot locate the platform within 60 seconds or if the mouse locates the platform within 60 seconds and stays on it for 10 seconds. After the test is over, stop the video, and put the mouse in its home cage under an infrared light.Finally perform the probe phase of the water maze test as previously described, without the platform. Next prepare the cylindrical tank for the forced swim test. Start the video recording, and gently put the mouse in the water in the center of the tank.Record the mouse for six minutes before putting the mouse back in its home cage. Review the video of the forced swim test. Measure the immobility time in the last five minutes of the video.This protocol was to study behavioral changes in mice after respiratory exposure to silica nanoparticles. The results in the open field test and the accelerating rotarod test demonstrated that silica nanoparticles did not affect locomotor or motor function in mice. Social interaction activity was affected after one-month exposure to silica nanoparticles, however the open field test, elevated plus maze test, and the forced swim test did not indicated any anxiety or depression after one month of exposure.A two-month exposure to silica nanoparticles did result in anxiety, as demonstrated by the results of the elevated plus maze test. Cognitive impairment was also detected in the water maze test after two months of exposure. This battery is designed for the comprehensive behavior analysis in neurodegeneration in the C57 mouse strain.It is most suitable for neurodegeneration-related longitudinal studies in toxicity of neurotoxin or potential risk factors, or drug development, which often features long-term administration and repeated testing.

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