Name: Assessment of Motor Balance and Coordination in Mice using the Balance Beam (Video) | JoVE Journal
Uploaded: 2011-03-10T15:00:00.000+00:00
Duration: 7 min 3 s
Description: Learn the balance beam walking assay to assess motor coordination and balance in mice. This sensitive method detects subtle deficits caused by brain injury, genetic changes, or drug treatments.

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5 minutes

Amber Southwell set up the apparatus and trained the authors in its use. Funding was provided by a grant from the NINDS to Paul Patterson.

1. This Protocol is Based on Those of Southwell et. al. (2009) and Carter et. al. (2001).
<ol>
 <li>The beam apparatus consists of 1 meter beams with a flat surface of 12 mm or 6 mm width) resting 50 cm above the table top on two poles. A black box is placed at the end of the beam as the finish point. (Plywood and other types of wood for the beams, poles, and box can be found at most hardware stores). Nesting material from home cages is placed in the black box to attract the mouse to the finish point. A lamp (with 60 watt light bulb) is used to shine light above the start point and serves as an aversive stimulus. The time to cross the center 80 cm is measured by two motion detectors (fabricated by Visopia): one at 0 cm that starts a timer and one at 80 cm that stops the timer. A nylon hammock (which can be obtained from a local fabric store) is stretched below the beam, ~7.5 cm above the table top, to cushion any falls. A video camera is set on a tripod to record the performance. The apparatus used in this demonstration was built on site.</li>
 <li>A cohort of at least 10 mice for each condition is recommended for this test. Mice are housed as groups, when possible, in a 12 hour light (7am-7pm)/dark cycle and acclimated to their cages in the behavioral facility for at least 2 weeks prior to the start of the test. Approximately 10 min prior to training/testing, the mice can be transported to the room containing the beam apparatus. </li>
 <li>On training days, each mouse crosses the 12 mm beam 3 times and then the 6 mm beam 3 times. Beams of wider widths (i.e. 28 mm) can be used for mice that are not able to cross the narrower ones or for larger mice. Mice are placed at one end of a beam and the time required to cross to the escape box at the other end (80 cm away) is measured by the motion detectors. The timer is started by the nose of the mouse entering the center 80 cm, and stopped when the animal reaches the end of the 80 cm. The mice rest for 10 min in their home cages between training sessions on the two beams. During this intertrial period, another 3-4 mice can be trained to make efficient use of time. C57BL/6 mice typically cross the beam with a minimal amount of stopping or stalling. However, when they do stall, sniff or look around without proceeding forward, the investigator should encourage the mouse to continue moving forward by prodding, poking, or pushing it from behind with gloved fingers. Once the mice are in the safe box, they are allowed some time (~15 secs) to rest there before the next trial. After each training session, the mice are returned directly to their home cages and taken back into their housing room. Of note, the mice may stall more if they are over-trained and become too familiar with the task and apparatus. On the other hand, training can be continued for additional days if the mice still do not traverse the entire beam successfully after 2 days of training. If treatments are applied to the mice, a baseline measure of performance should be obtained prior to treatment.</li>
 <li>On the test day, times to cross each beam are recorded. Two successful trials in which the mouse did not stall on the beam are averaged. Video recordings can be used for finer analysis of slipping and other observable motor deficits. An alternative way to assess beam walking is by a neurological scoring system1,2, which rates the inability to cross the beam as a score of "1" and the ability to traverse the beam normally with both paws on the beam surface and fewer than two foot slips as a "7." A slip is defined as the foot coming off the top of the beam. In this indexing system, a score of "3" would correspond to crossing the beam by dragging limbs. An animal that scores 1 on two trials is not retested. Description of performance on the beam related to other scores can be found in Feeney et al., (1982) and Carter et al., (2001).</li>
 <li>The beams and box are cleaned of mouse droppings and wiped with towels soaked with 70% ethanol and then water before the next beam is placed on the apparatus. </li>
</ol>
2. Representative Results:
<img src="/files/ftp_upload/2376/2376fig1.jpg" alt="Average time to cross beam graph; male/female C57Bl6; 12mm/6mm beam comparison; behavioral study." data-altupdatedat="1788235205000" data-alt="Figure 1"> 
Figure 1. Typical cross times of male and female C57Bl6 on 6mm and 12mm beams. A cohort of 9 to 11 week old C57BL/6-NCRL WT mice were tested on both beams after 2 days of training. Average times for crossing the 12 mm beam were 4.6 ± 0.4 sec for males (n=14) and 3.3 ± 0.3 sec for females (n=11). Times on the 6 mm beam were 6.8 ± 0.7 sec for males (n=13) and 5.9 ± 0.5 sec for females (n=10) (errors are reported as S.E.M.).

<ol><li>Feeney, D. M., Gonzalez, A., Law, W. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Amphetamine%2C+haloperidol%2C+and+experience+interact+to+affect+rate+of+recovery+after+motor+cortex+injury%5BTitle%5D)+AND+%22Science%22%5BJournal%5D)">Amphetamine, haloperidol, and experience interact to affect rate of recovery after motor cortex injury.</a> Science. 217, 855-857 (1982).</li>
<li>Carter, R. J., Morton, J., Dunnett, S. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Motor+coordination+and+balance+in+rodents%5BTitle%5D)+AND+%22Curr+Protoc+Neurosci%22%5BJournal%5D)">Motor coordination and balance in rodents.</a> Curr Protoc Neurosci. Chapter 8, (2001).</li>
<li>Wallace, J. E., Krauter, E. E., Campbell, B. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Motor+and+reflexive+behavior+in+the+aging+rat%5BTitle%5D)+AND+%22J+Gerontol%22%5BJournal%5D)">Motor and reflexive behavior in the aging rat.</a> J Gerontol. 35, 364-370 (1980).</li>
<li>Brooks, S. P., Dunnett, S. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Tests+to+assess+motor+phenotype+in+mice%3A+a+user%27s+guide%5BTitle%5D)+AND+%22Nat+Rev+Neurosci%22%5BJournal%5D)">Tests to assess motor phenotype in mice: a user's guide.</a> Nat Rev Neurosci. 10, 519-529 (2009).</li>
<li>Fox, G. B., Fan, L., Levasseur, R. A., Faden, A. I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Sustained+sensory%2Fmotor+and+cognitive+deficits+with+neuronal+apoptosis+following+controlled+cortical+impact+brain+injury+in+the+mouse%5BTitle%5D)+AND+%22J+Neurotrauma%22%5BJournal%5D)">Sustained sensory/motor and cognitive deficits with neuronal apoptosis following controlled cortical impact brain injury in the mouse.</a> J Neurotrauma. 15, 599-614 (1998).</li>
<li>Buccafusco, J. J. <a target="_blank" href="http://scholar.google.com/scholar?hl=en&safe=off&q=author%3aBuccafusco%2C+JJ+%22Methods+of+behavioral+analysis+in+neuroscience%22">Methods of behavioral analysis in neuroscience</a>. , 2nd edn, CRC Press. (2009).</li>
<li>Southwell, A. L., Ko, J., Patterson, P. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Intrabody+gene+therapy+ameliorates+motor%2C+cognitive%2C+and+neuropathological+symptoms+in+multiple+mouse+models+of+Huntington%27s+disease%5BTitle%5D)+AND+%22J+Neurosci%22%5BJournal%5D)">Intrabody gene therapy ameliorates motor, cognitive, and neuropathological symptoms in multiple mouse models of Huntington's disease.</a> J Neurosci. 29, 13589-13602 (2009).</li>
<li>Carter, R. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Characterization+of+progressive+motor+deficits+in+mice+transgenic+for+the+human+Huntington%27s+disease+mutation%5BTitle%5D)+AND+%22J+Neurosci%22%5BJournal%5D)">Characterization of progressive motor deficits in mice transgenic for the human Huntington's disease mutation.</a> J Neurosci. 19, 3248-3257 (1999).</li>
<li>Gulinello, M., Chen, F., Dobrenis, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Early+deficits+in+motor+coordination+and+cognitive+dysfunction+in+a+mouse+model+of+the+neurodegenerative+lysosomal+storage+disorder%2C+Sandhoff+disease%5BTitle%5D)+AND+%22Behav+Brain+Res%22%5BJournal%5D)">Early deficits in motor coordination and cognitive dysfunction in a mouse model of the neurodegenerative lysosomal storage disorder, Sandhoff disease.</a> Behav Brain Res. 193, 315-319 (2008).</li>
<li>Stanley, J. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((The+mouse+beam+walking+assay+offers+improved+sensitivity+over+the+mouse+rotarod+in+determining+motor+coordination+deficits+induced+by+benzodiazepines%5BTitle%5D)+AND+%22J+Psychopharmacol%22%5BJournal%5D)">The mouse beam walking assay offers improved sensitivity over the mouse rotarod in determining motor coordination deficits induced by benzodiazepines.</a> J Psychopharmacol. 19, 221-227 (2005).</li>
</ol>

Experiments on animals were performed in accordance with the guidelines and regulations set forth by the Caltech Institutional Animal Care and Use Committee (IACUC).

Performance on the balance beam is a useful measure of fine coordination and balance and has been validated by previous work. The beam test can detect motor deficits due to age, central nervous syst` lesions, and genetic and pharmacological manipulations in young and older rodents2,3,4. Mice with cortical impact lesions often exhibit contralateral slipping on the beam5,6. The beam test has also been used to differentiate the motor skills of wild-type (WT) and Huntington disease (HD) mouse models ...

<table><tbody><tr><td>Beams: 1-m strips of smooth wood with small, medium, and large square (i.e., 6, 12, and 28 mm wide) cross-sections</td><td></td><td></td><td></td></tr><tr><td>Support stand to hold up the start section of the raised beam (&amp;#8764;1.5-cm cross-section, 50 cm high)</td><td></td><td></td><td></td></tr><tr><td>Goal box (20 cm on each side) mounted on a support stand (3 cm cross-section, 50 cm high)</td><td></td><td></td><td></td></tr><tr><td>Hammock-&amp;#8764;7.5 cm above tabletop</td><td></td><td></td><td></td></tr><tr><td>60-watt (W) desk lamp</td><td></td><td></td><td></td></tr><tr><td>Video camera, tripod, and blank video tapes</td><td></td><td></td><td></td></tr><tr><td>Ethanol, paper towels, cleaning materials</td><td></td><td></td><td></td></tr></tbody></table>

assessment of motor balance and coordination in mice using the balance beam

Assessment of Motor Balance and Coordination in Mice using the Balance Beam

Brain injury, genetic manipulations, and pharmacological treatments can result in alterations of motor skills in mice. Fine motor coordination and balance can be assessed by the beam walking assay. The goal of this test is for the mouse to stay upright and walk across an elevated narrow beam to a safe platform. This test takes place over 3 consecutive days: 2 days of training and 1 day of testing. Performance on the beam is quantified by measuring the time it takes for the mouse to traverse the beam and the number of paw slips that occur in the process. Here we report the protocol used in our laboratory, and representative results from a cohort of C57BL/6 mice. This task is particularly useful for detecting subtle deficits in motor skills and balance that may not be detected by other motor tests, such as the Rotarod.

Deficits in fine motor coordination can be assessed with the balance beam test. Performance on the beam is quantified by the speed at which the beam is traversed and the number of times the mouse slips on the beam.

Brain injury, genetic manipulations, and pharmacological treatments can result in alterations of motor skills in mice.  Fine motor coordination and ...

The overall goal of this procedure is to assess motor coordination and balance in mice. The first step is to assemble a balance beam with motion detector's position to record the latency of beam crossing to begin training maer placed on the starting point of the balance beam and allowed to cross to a safe platform. It is sometimes necessary to encourage the animal along the beam toward the end point.After an inter trial interval of 10 minutes, maer then trained on a narrower beam to allow for greater sensitivity. The final step of the procedure is to test the trained animals on beams of different widths. Ultimately, results can be obtained that show deficits in motor coordination and balance.Hi, I am Tim Luong and I'm an MD PhD candidate in the biology department at the California Institute of Technology. I'm Holly Carlisle and I'm a postdoc at the California Institute of Technology in the biology department, and today we're going to be showing you how to assess balance and motor coordination in mice using the balance beam test. Now, the main advantage of this technique compared to other methods such as the rotor rod, is that the balance beam test tends to be more sensitive to even subtle deficits in motor coordination.This method can help answer key questions in the behavioral field, such as whether our motor coordination and balance are affected in certain neurological disease models and whether our motor deficits are alleviated by particular treatments. The purpose of this test is to measure the amount of time it takes for a mouse to cross the beam without stopping or stalling and without prompting from the investigator. In order to get a clean run on test day, we train for two days prior to testing.Visual demonstration of this method is helpful as the way the mice are handled and aided across the beam may be difficult for those less familiar with behavioral testing. So let's get started. This protocol is based on those of Southwell, etal and Carter etal.The apparatus is assembled by resting a beam that is one meter long between two 50 centimeter high poles. The beams have a flat surface of six or 12 millimeters wide. Beams of greater widths can be used for larger mice and those not able to cross the narrower sizes.A black box is placed at the end of the beam and nesting material is placed inside the black box to encourage the mouse towards the finishing point, illuminate the starting point with a lamp to serve as an aversive stimulus. Motion detectors are placed at zero and 80 centimeters to record the latency of beam crossing, stretch a nylon hammock below the beam and approximately 7.5 centimeters above the tabletop to cushion any falls. The mice used in this procedure are housed on a 12 hour light dark cycle and have been acclimated for two weeks prior to testing.Introduced the animals to the test room approximately 10 minutes before beginning the experiment and use a cohort of at least 10 mice per test condition for the best results. To begin training, place a mouse on the start location of a 12 millimeter wide beam. The timer starts when the nose of the mouse enters the center 80 centimeters and stops when the animal reaches the end of the 80 centimeters.Mice should cross the beam with a minimal amount of stopping or stalling. If the animal stops moving forward, encourage them to continue by gently prodding them from behind. Once the animal has successfully traversed the beam, remove it from the safe box and place it back into the home cage for 10 minutes before beginning the next training session.The balance beam and box are cleaned and wiped with 70%ethanol before introducing the next mouse. Each mouse is trained on the 12 millimeter wide beam three times followed by training on the six millimeter wide beam. An additional three times over training may increase the occurrence of stalling as the animal becomes more familiar with the task and apparatus.On the other hand, if mice fail to traverse the entire beam successfully after two days of training, additional training may be required. Be sure to record baseline measurements before the animals receive any treatment. After training is complete for the day, return mice to their home cages into the holding facility.On the day of testing, bring the animals into the room 10 minutes before beginning the experiment as shown previously, place a mouse in the start location and record the latency for the animal to traverse the center 80 centimeters of the beam. The data are reported as the average of two successful trials in which the mouse crossed directly to the end without stopping and without prodding by the investigator. Video recordings can be used for finer analysis of slipping and other observable motor deficits.In this example, nine to 11 week old C 57 black six mice were tested on 12 millimeter and six millimeter wide beams. The average time to cross the 12 millimeter wide beam was 4.6 plus or minus 0.4 seconds for males, and 3.3 plus or minus 0.3 seconds for females times on the six beam were 6.8 plus or minus 0.7 seconds for males and 5.9 plus or minus 0.5 seconds. For females.Balance beam performance may vary depending on the age and strain of mice. The C 57 black six mice used in this example are typically active animals, a few slips, but no falls were observed for these mice and genetically or pharmacologically manipulated animals, slips and falls may become more frequent and can be quantified. Impaired mice may clinging onto the side of the beam, which increases their time to cross While attempting this procedure.It's important to be persistent and consistent in prodding the mice to move forward if they stall or get distracted on the beam. After watching this video, you should have a good understanding of how to train mice to cross the balance beam and to assess their balance and coordination. Thanks for watching and I hope you found this helpful.

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Learn the balance beam walking assay to assess motor coordination and balance in mice. This sensitive method detects subtle deficits caused by brain injury, genetic changes, or drug treatments.

Assessment of Motor Balance and Coordination in Mice using the Balance Beam

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