We would like to thank The New Hope Resarch&#160;Foundation for their continued support and funding of our research. We would also like to thank IITC for partial financial support for this work.

All protocols herein described were performed in accordance with, and were approved by, the Queen&#39;s University Animal Care Committee. The frequency of behavioral testing can be weekly or monthly, or a combination of the two (i.e., weekly when one expects to see the biggest difference, and monthly elsewhere).
NOTE: Murine behavioral studies are largely sex/age/strain- dependent, and therefore it is essential to collect the proper time points and to remain consistent across experimental groups. Environmental factors (e.g., scents, noise, temperature, humidity lighting, etc.) greatly affect behavior and anxiety in mice. Try to reduce these environmental factors as much as possible.
1. Open Field Test
NOTE: Prior to behavioral testing, the animals should be habituated to the room where the testing is to take place for 10 - 15 min prior to the beginning of testing. Ensure that proper personal protective equipment and all local facility guidelines are followed for the behavioral testing protocols.
NOTE: No training prior to the test.
<ol>
	<li>Ensure that the computer and OFT program are on, running, and properly connected to the camera or open field box. Click on the smartware program, the open field icon, and &#34;OK.&#34; Chose the &#34;digital analog converter&#34; source from the camera options. Click on &#34;detection&#34; and make sure that the whole open field space is visualized and that the dimensions are 30 cm x 30 cm. 
	NOTE: Equipment layout may vary depending on space constraints, but ensure that the computer and open field apparatus are near to each other in a well-lit room for proper video capture.</li>
	<li>Click &#34;calibration,&#34; &#34;snapshot,&#34; and then &#34;OK&#34; to make sure that the mouse is being detected within the confines of the open field. Click &#34;time&#34; to pre-set an amount of time to test the mice; in this case, set 5 min.</li>
	<li>Enter the subjects by clicking the &#34;subjects&#34; icon. Click &#34;scheduler&#34; and highlight all test subjects and then click the double green arrows and &#34;phase one, session one;&#34; this will bring in all of the subjects in order to begin collecting data.</li>
	<li>Place a mouse in the center of the open field and click the &#34;start&#34; icon to prompt the program to begin recording. Test only one mouse at a time.</li>
	<li>Once 5 min is complete, the program will automatically move to the next subject. In this case, remove the mouse that has completed the test and place the next subject into the open field. Click &#34;start.&#34;</li>
	<li>Wipe down the equipment. Repeat steps 1.3 - 1.4 for the remaining test subjects.</li>
	<li>In order to analyze the data once all the subjects have gone through testing, click on the &#34;analyze&#34; icon at the top right of the screen. Click &#34;analyze&#34; again and then &#34;summary report.&#34; Optionally, export the data onto a USB.</li>
</ol>
2. Mesh Test
NOTE: The surface area and size of the mesh on the MT may influence performance. On the MT, if a mouse falls off within the first 10 s during the first 2 collection time points, they will be put back on the mesh immediately for a second attempt. The mesh used herein was a wire mesh, with the mesh holes approximately 1.5 mm in diameter.
NOTE: Running these tests at the same time of day is recommended, since physiological and biochemical parameters fluctuate throughout the day. Bodyweight may influence performance in this test and therefore must be taken into consideration.
<ol>
	<li>Place a mouse in the center of a mesh and invert the mesh at least 20 cm above a clear container. Once inverted, start the timer.</li>
	<li>Time and record the time of latency to fall. If the mouse does not fall within 60 s, give it a score of 60 s and conduct no further trials. Allow the mouse to rest for at least 5 min. This is the end of trial 1.</li>
	<li>Wipe down the equipment and during the 5-min inter-trial interval, test a second mouse, if required. Repeat steps 2.1 - 2.2 for trials 2 and 3, if necessary (as long as the mouse does not score 60 s, it should undergo three trials).</li>
</ol>
3. Rotarod Test
NOTE: The surface and diameter of the rod on the RT may influence performance. On the RT, if a mouse falls off within the first 10 s during the first 2 collection time points, put it back on the rod immediately for a second attempt.
NOTE: Running these tests at the same time of day is recommended, since physiological and biochemical parameters fluctuate throughout the day. Bodyweight may influence performance in this test and therefore must be taken into consideration.
<ol>
	<li>Turn on the IITC rotarod and computer. Ensure that they are properly connected for data recording.</li>
	<li>Make sure that the program is running the accelerated protocol. Set the desired length of the rotarod test to xxx (the valid range is 000 through 999); in this case, press 300*. Set the number of lanes to be used to xxx (the valid range is 001 through 005); in this case, press 005*. Set the starting speed to xxx rpms (enter 3005 for 5 rpms; the valid range is 001 through 045); in this case, press 004*.</li>
	<li>Set the top speed to xxx rpms (enter 4,030 for 30 rpms; the valid range is 001 through 045); in this case, press 040*. Set the ramp speed to xxx in s (enter 5,030 for a 30-s ramp to top speed; the valid range is 000 through 999); in this case press 300*. Set the unit to mouse mode (enter 6001 for mouse forward and 6002 for mouse reverse); in this case, press 6001*.</li>
	<li>Ensure that all magnetized lanes are balanced before beginning the trial. Place one mouse in each lane of the rotarod.</li>
	<li>Press start to initiate the program. As the mice fall, remove them from their lanes promptly to ensure that they do not &#34;trip&#34; the other lanes.</li>
	<li>Allow the mice to rest for a 10-min inter-trial interval. This is the end of trial 1. During the inter-trial interval, wipe down the equipment and begin the trial for the next set of mice.</li>
	<li>Repeat steps 4 - 6 as necessary. If a mouse remains on the rotarod for the full 5-min trial, no further trials are required for that mouse.</li>
</ol>

<ol>
	<li>Osmon, K. J., 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=Systemic+Gene+Transfer+of+a+Hexosaminidase+Variant+Using+an+scAAV9.47+Vector+Corrects+GM2+Gangliosidosis+in+Sandhoff+Mice.">Systemic Gene Transfer of a Hexosaminidase Variant Using an scAAV9.47 Vector Corrects GM2 Gangliosidosis in Sandhoff Mice.</a> Human Gene Therapy. 27 (7), 497-508 (2016).</li><li>Brooks, S. P., Dunnett, S. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Tests+to+assess+motor+phenotype+in+mice:+a+user's+guide.">Tests to assess motor phenotype in mice: a user's guide.</a> Nature Reviews, Neuroscience. 10, 519-529 (2009).</li><li>Ma, M., Basso, D. M., Walters, P., Stokes, B. T., Moser, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Behavioural+and+histological+outcomes+following+graded+spinal+cord+contusion+injury+in+the+C57Bl/6+mouse.">Behavioural and histological outcomes following graded spinal cord contusion injury in the C57Bl/6 mouse.</a> Exp. Neurol. 169, 239-254 (2001).</li><li>Menalled, L. B., 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=Early+motor+dysfunction+and+striosomal+distribution+of+huntingtin+microaggregates+in+Huntington's+disease+knock-in+mice.">Early motor dysfunction and striosomal distribution of huntingtin microaggregates in Huntington's disease knock-in mice.</a> J.Neurosci. 22, 8266-8276 (2002).</li><li>Grusser, C., Grusser-Cornehls, U. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Improvement+in+motor+performance+of+Weaver+mutant+mice+following+lesion+of+the+cerebellum.">Improvement in motor performance of Weaver mutant mice following lesion of the cerebellum.</a> Behav. Brain Res. 97, 189-194 (1998).</li><li>De Leonibus, E., 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=Spatial+deficits+in+a+mouse+model+of+Parkinson's+disease.">Spatial deficits in a mouse model of Parkinson's disease.</a> Psychopharmacology (Berl). 194, 517-525 (2007).</li><li>Morris, M., Koyama, A., Masliah, E., Mucke, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Tau+Reduction+Does+Not+Prevent+Motor+Deficits+in+Two+Mouse+Models+of+Parkinson's+Disease.">Tau Reduction Does Not Prevent Motor Deficits in Two Mouse Models of Parkinson's Disease.</a> PLoS ONE. 6 (12), 1-7 (2011).</li><li>Abo-ouf, A., 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=Deletion+of+tumour+necrosis+factor-&#945;+ameliorates+neurodegeneration+in+Sandhoff+disease+mice.">Deletion of tumour necrosis factor-&#945; ameliorates neurodegeneration in Sandhoff disease mice.</a> Human Molecular Genetics. , 1-16 (2013).</li><li>Dumont, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Behavioral+Phenotyping+of+Mouse+Models+of+Neurodegeneration.">Behavioral Phenotyping of Mouse Models of Neurodegeneration.</a> Neurodegeneration : Methods and Protocols, Methods in Molecular Biology. 793, 229-237 (2011).</li><li>Cabe, P. A., Tilson, H. A., Mitchell, C. L., Dennis, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+simple+recording+grip+strength+device.">A simple recording grip strength device.</a> Pharmacol. Biochem. 8, 101-102 (1978).</li><li>Barn&#275;oud, P., Loliver, J., Sanger, D., Moser, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Quantitative+motor+assessment+in+FALS+mice:+a+longitudinal+study.">Quantitative motor assessment in FALS mice: a longitudinal study.</a> Neuroreport. 8, 2861-2865 (1997).</li><li>Lui, Y., 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=Mouse+model+of+GM2+activator+deficiency+manifests+cerebellar+pathology+and+motor+impairment.">Mouse model of GM2 activator deficiency manifests cerebellar pathology and motor impairment.</a> Proc. Natl. Acad. Sci. 94, 8133-8143 (1997).</li><li>Sango, K., 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=Mouse+models+of+Tay-Sachs+and+Sandhoff+diseases+differ+in+neurologic+phenotype+and+ganglioside+metabolism.">Mouse models of Tay-Sachs and Sandhoff diseases differ in neurologic phenotype and ganglioside metabolism.</a> Nature Genetics. 11 (2), 170-176 (1995).</li><li>Jones, B. J., Roberts, D. 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+rotarod+suitable+for+quantitative+measurements+of+motor+inco-ordination+in+na&#239;ve+mice.">A rotarod suitable for quantitative measurements of motor inco-ordination in na&#239;ve mice.</a> Nauyn-Schmiedebergs Arch. Pharmacol. 259 (2), 211 (1968).</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=Measuring+the+Strength+of+Mice.">Measuring the Strength of Mice.</a> Journal of Visualized Experiments JoVE. (76), (2013).</li></ol>

The authors have nothing to disclose.

For all tests, Clean and disinfect the surface to minimize contagions and distractors to the mice.&#160;Also, make certain that the mice are given sufficient rest periods between trials and between behavioral testing when using the OFT, RR, and MT.
In the OFT, it is critical that each mouse is placed in the middle of the open field and that the program is started upon the release of the mouse into the center. Be sure that there are no hand movements the sensors may register upon starting the p...

The goal of behavioral testing is to measure phenotypic characterization, whether in studies for a new mouse model of diseases affecting the central nervous system or for a trial of a new treatment. There are a variety of behavioral tests to choose from. The rationale for using a combination of behavioral tests is often to fully assess the pre-clinical treatment outcomes. The battery of behavioral testing herein includes the open field test (OFT), the mesh test (MT), and the rotarod test (RR).
The OFT is used as a measure of general locomotor activity1,2,3,4,5,6,7. This assessment allows the mouse to move freely in an open field, and the measurements taken are the movements of the mouse over the course of the behavioral trial. The quantitative analyses made using an OFT include rearing, distance moved, time spent moving (e.g., walking and running), and changes in activity over time (e.g., jumping and slower/hyperactive movements), which is why it is a useful and thorough analysis of rodent behavior.
The MT is used to assess the strength of the mice1,2,8,9,10,11. The use of all four limbs to hang off of a wire grid mesh allows for the non-invasive measurement of the ability of the mice to show sustained limb tension while opposing the effects of gravity. This assessment is done by placing a mouse on an inverted mesh and timing its latency to fall. This test assesses the muscular strength of the mouse by measuring how long it can hold itself onto the inverted screen before falling off.
The RR assesses the coordination of a mouse while using the accelerating protocol1,2,7,8,9,12,13. The rotarod apparatus consists of a cylindrical beam that rotates in the air starting at 4 rpm. The mouse can learn this motor coordination skill and is then challenged by slow, incremental increases up to 40 rpm. Classically, the constant speed protocol is a measure of muscular strength, whereas the accelerating protocol for the RR assesses coordination, endurance, and muscular strength1,2,7,14.
The advantages of these three behavioral tests are that they are widely accepted throughout the rodent behavioral testing community2,7,8,9, and when taken not just as the sum of the individual tests, but collectively, they can be used as a global assessment of phenotypic behavior when investigating the different outcomes of experimental treatments. The OFT is one of the only tests for overall locomotor activity2. The RR is also one of the only ways to assess coordination2, other than to use gait analysis software, which will not be discussed here. There are a few ways in which the muscular strength of a mouse can be assessed; however, using the MT, or a wire hang test, has been prevalently used in the literature and is accepted as a reliable method for testing muscular strength2,8,9,10,11. Many other techniques for measuring general locomotion and coordination are extremely time consuming and may not produce reliable data (e.g., manually timing mouse movement or manually painting paws for gait analysis). The automated systems for the OFT and the RR increase the reliability and validity of the data produced and save time and effort.
For example, given the neurodegeneration of the Sandhoff disease mouse model, when performing this battery of tests, the OFT would be the first of these tests used to assess the motor abnormalities that are usually seen in this mouse model starting at 3 months of age. Deficits in motor coordination should also become apparent beginning at 12 weeks of age. Deficits in muscular strength should begin around 14 weeks of age, but they ultimately become much more significant by 16 weeks.
These tests are useful when studying diseases that cause significant motor impairments, including neurodegenerative diseases9 such as Sandhoff disease1,8,13, Parkinson&#39;s disease7, Huntington&#39;s disease, multiple sclerosis. The tests can also be used to study aging and to make comparative geriatric assessments or measurements of health. Gait disturbances and changes in locomotion and balance are seen in many of these disease states. However, the usefulness of these tests is not limited to diseases with prevalent neuromuscular symptoms.
Please note that there have been no confounding variables noted with respect to motivation or the novelty of any of the described behavioral testing measures. It is imperative for certain disease states and pathologies that the researchers acquire preliminary behavioral data before the mouse shows any phenotypic symptoms; the researcher should then periodically (weekly) ensure that all motor impairments are accounted for. Many diseases have a quick onset of phenotypic symptoms, and without weekly testing, these symptoms would not be accounted for.

<table><tbody><tr><td>IITC Life Sciences Rotarod with Series 8 computing software&amp;nbsp;</td><td>IITC Life Sciences</td><td>755</td><td></td></tr><tr><td>Square Open Field 45 x 45 x 40 cm (grey color). Other floor color available: white, black.</td><td>Harvard Aparatus (PanLab)</td><td>760190</td><td></td></tr><tr><td>&amp;nbsp;Open Field Preconfigured Module</td><td>Harvard Aparatus (PanLab)</td><td>760688</td><td></td></tr><tr><td>Smart V3.0 Software Platform&amp;nbsp;</td><td>Harvard Aparatus (PanLab)</td><td>760681</td><td></td></tr><tr><td>Video converter (analog/digital)</td><td>Harvard Aparatus (PanLab)</td><td>760262</td><td></td></tr><tr><td>Roof fixing sys for camera</td><td>Harvard Aparatus (PanLab)</td><td>760277</td><td></td></tr><tr><td>Analog Sony Camera &amp;amp; lenseNTSC</td><td>Harvard Aparatus (PanLab)</td><td>760505</td><td></td></tr></tbody></table>

battery of behavioral tests assessing general locomotion, muscular strength, and coordination in mice

Behavioral testing is used in pre-clinical trials to assess the phenotypic effects and outcomes that a particular disease or treatment has on the animal&#39;s wellbeing and health. There are numerous behavioral tests that may be applied. We selected a test for general locomotion, the open field test (OFT); a test for muscular strength, the mesh test (MT); and a test for coordination, the rotarod test (RR). Testing can be accomplished on a weekly or monthly basis. As a test for general locomotion, the OFT works by objectively monitoring movement parameters while the mouse is in an open field apparatus. The field is generally a 2&#39; x 2&#39; box, and the movements are recorded through laser sensing or through video capture. The mouse is placed in the center of the open field and allowed to move freely for the test. The MT uses the latency for a mouse to fall off an inverted screen as a measure of muscular strength. A mouse is placed on a screen, which is inverted over a clear box, and is timed for their latency to fall. Three trials are performed, with the best of the three trials scored for that day. A score of 60 s is the maximum time a mouse is left inverted. Mice are given a 5-min rest period between mesh test trials. Lastly, an accelerated protocol on the RR assesses motor coordination and endurance. During a trial, a mouse walks on a rotating rod as it increases in speed from 4 rpm to 40 rpm over 5 min. The trial ends when the mouse touches the magnetized pressure sensor upon falling. Each mouse undergoes three trials, and the best trial is scored for that day. This combined behavioral data allows for the global assessment of mobility, coordination, strength, and movement of the test animals. At least two out of the three behavioral testing measures must show improvement for an animal to qualify as having overall improved motor function.

The study animals may undergo the behavioral testing paradigm weekly or monthly to accommodate the necessary time points required for analysis. See Figure 1 for examples on behavioral testing timing. In the representative data below, taken from Osmon et al. (2016), the mice underwent the battery of behavioral testing monthly, beginning at 12 weeks of age, until their humane endpoint around 40 weeks of age. The timing of behavioral testing is sometimes critical fo...

Watch this Scientific Journal Video about Battery of Behavioral Tests Assessing General Locomotion, Muscular Strength, and Coordination in Mice at JoVE.com

Battery of Behavioral Tests Assessing General Locomotion, Muscular Strength, and Coordination in Mice

Behavioral testing is used in pre-clinical trials to assess the phenotypic effects of a disease or treatment on an animal's wellbeing. In order to globally assess motor functioning, we selected tests for general locomotion, muscular strength, and coordination: the open field test, the mesh test, and the rotarod test, respectively.

Behavioral testing is used in pre-clinical trials to assess the phenotypic effects and outcomes that a particular disease or treatment has on the ...

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Universidad de Cartagena UDEC

Research

JoVE Journal

Behavior

13.5K Views.  Queen's University. Behavioral testing is used in pre-clinical trials to assess the phenotypic effects of a disease or treatment on an animal's wellbeing. In order to globally assess motor functioning, we selected tests for general locomotion, muscular strength, and coordination: the open field test, the mesh test, and the rotarod test, respectively. 

Video: Battery of Behavioral Tests Assessing General Locomotion, Muscular Strength, and Coordination in Mice

Assessment of Sensorimotor Function in Mouse Models of Parkinson's Disease

Sensitive and reliable behavioral outcome measures are essential to the evaluation of potential therapeutic treatments in preclinical trials for many neurodegenerative diseases. In Parkinson's disease, sensorimotor tests sensitive to varying degrees of nigrostriatal dysfunction are fundamental for testing the efficacy of potential therapeutics. Reliable and quite elegant sensorimotor measures exist for rats, however many of these tests measure sensorimotor asymmetry within the rat and are not entirely suitable for the newer genetic mouse models of PD. We have put together a battery of sensorimotor tests inspired by the sensitive tests in rats and adapted for mice. The test battery highlighted in this study is chosen for a) its sensitivity in a wide variety of mouse models of PD, b) its ease in implementing into a study, and c) its low expense. These tests have proven useful in characterizing novel genetic mouse models of PD as well as in testing potential disease-modifying therapies.

In Parkinson's disease and movement disorders in general, sensitive and reliable behavioral assays are essential for testing novel potential therapeutics. Here, we describe a manageable battery of sensorimotor tests for mice that are sensitive to varying degrees of injury to the nigrostriatal system and useful for preclinical studies.

Sensitive and  reliable behavioral outcome measures are essential to the evaluation of  potential therapeutic treatments in preclinical trials for many  ...

Measuring the Strength of Mice

Kondziela7 devised the inverted screen test and published it in 1964. It is a test of muscle strength using all four limbs. Most normal mice easily score maximum on this task; it is a quick but insensitive gross screen, and the weights test described in this article will provide a finer measure of muscular strength.
There are also several strain gauge-based pieces of apparatus available commercially that will provide more graded data than the inverted screen test, but their cost may put them beyond the reach of many laboratories which do not specialize in strength testing. Hence in 2000 a cheap and simple apparatus was devised by the author. It consists of a series of chain links of increasing length, attached to a "fur collector" a ball of fine wire mesh sold for preventing limescale build up in hard water areas. An accidental observation revealed that mice could grip these very tightly, so they proved ideal as a grip point for a weight-lifting apparatus. A common fault with commercial strength meters is that the bar or other grip feature is not thin enough for mice to exert a maximum grip. As a general rule, the thinner the wire or bar, the better a mouse can grip with its small claws.
This is a pure test of strength, although as for any test motivational factors could potentially play a role. The use of scale collectors, however, seems to minimize motivational problems as the motivation appears to be very high for most normal young adult mice.

Deficits in muscular strength occur in many clinical conditions such as motor neuron disease. The inverted screen and weight lifting tests described here measure strength in mice almost exclusively, with minimal influence of factors such as coordination.

Kondziela7 devised the inverted screen test and published it in 1964. It is a test of muscle strength using all four limbs. Most normal mice easily score ...

Medicine

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 ...

Neuroscience

Measuring Motor Coordination in Mice

Mice are increasingly being used in behavioral neuroscience, largely replacing rats as the behaviorist's animal of choice. Before aspects of behavior such as emotionality or cognition can be assessed, however, it is vital to determine whether the motor capabilities of e.g. a mutant or lesioned mouse allow such an assessment. Performance on a maze task requiring strength and coordination, such as the Morris water maze, might well be impaired in a mouse by motor, rather than cognitive, impairments, so it is essential to selectively dissect the latter from the former. For example, sensorimotor impairments caused by NMDA antagonists have been shown to impair water maze performance2. 
Motor coordination has traditionally been assessed in mice and rats by the rotarod test, in which the animal is placed on a horizontal rod that rotates about its long axis; the animal must walk forwards to remain upright and not fall off. Both set speed and accelerating versions of the rotarod are available.
The other three tests described in this article (horizontal bar, static rods and parallel bars) all measure coordination on static apparatus. The horizontal bar also requires strength for adequate performance, particularly of the forelimbs as the mouse initially grips the bar just with the front paws. 
Adult rats do not perform well on tests such as the static rods and parallel bars (personal observations); they appear less well coordinated than mice. I have only tested male rats, however, and male mice seem generally less well coordinated than females. Mice appear to have a higher strength:weight ratio than rats; the Latin name, Mus musculus, seems entirely appropriate. The rotarod, the variations of the foot fault test12 or the Catwalk (Noldus)15 apparatus are generally used to assess motor coordination in rats.

Protocols are presented for two established motor coordination tasks, the accelerating rotarod and horizontal bar, also two tests developed in Oxford recently, the static rods and parallel bars. These tests can detect motor impairments potentially of interest in their own right, as well as being possible variables in tests of other areas of behavior.

Mice are increasingly being used in behavioral neuroscience, largely replacing rats as the behaviorist's animal of choice. Before aspects of behavior such ...

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.

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 ...

Assessing Motor Performance in TH-Related Murine Models Using the Ladder Beam Test

A Versatile, Behavioral Method to Investigate Thyroid Hormone Effects on Cerebellar Function

Thyroid hormone (TH) action is essential during the development of the central nervous system, including the cerebellum. In case of TH deficiency in early life such as congenital hypothyroidism, patients display neurological disorders such as cognitive retardation and motor deficits. There are various studies using mouse models with tissue- or cell-specific TH deficiency to investigate the role of TH in the cerebellum. Compared to generalized congenital hypothyroid mice, cerebellar cell-specific TH-deficient mice display milder and subtler ataxic features, making the assessment of motor function difficult when using conventional tests such as the rotarod test. 
Due to the need for an alternative tool to assess motor function in TH-related animal models, we developed a versatile behavioral method called the "ladder beam test," in which we can design the various ladder tests depending on the severity of ataxia in model mice. We utilized transgenic mice expressing a dominant-negative TH receptor specifically in the cerebellar Purkinje cell, a sole output neuron in the cerebellar cortex modulating motor performance. The newly-built ladder beam test successfully detected robust impairments in motor performance in the transgenic mice at a greater level compared to the rotarod test. Disruption of motor learning was also detected in the ladder beam test but not in the rotarod test. The protocol with this novel behavioral apparatus can be applied to other animal models that may show mild ataxic phenotype to examine subtle changes in cerebellar function.

Author Spotlight: Accurately Assessing Thyroid Hormone-Driven Motor Alterations in Mouse

Here we present a protocol for a versatile behavior test developed recently, the ladder beam test. This test has the advantage of detecting subtle cerebellar ataxia caused by a defect of thyroid hormone action in the central nervous system over the conventional behavior tests assessing motor performance.

Thyroid hormone (TH) action is essential during the development of the central nervous system, including the cerebellum. In case of TH deficiency in early ...

Validating a Murine Model of Angelman Syndrome Using Behavioral Tests

Behavioral Characterization of an Angelman Syndrome Mouse Model

This manuscript describes a battery of behavioral tests available to characterize Angelman syndrome (AS)-like phenotypes in an established murine model of AS. We use the rotarod learning paradigm, detailed gait analysis, and nest building test to detect and characterize animal motor impairments. We test animal emotionality in the open field and elevated plus maze tests, as well as the affect in the tail suspension test. When AS mice are tested in the open field test, the results should be interpreted with care, since motor dysfunctions influence mouse behavior in the maze and alter activity scores.
The reproducibility and effectiveness of the presented behavioral tests has already been validated in several independent Uba3a mouse lines with different knockout variants, establishing this set of tests as an excellent validation tool in AS research. Models with the relevant construct and face validity will warrant further investigations to elucidate the pathophysiology of the disease and grant the development of causal treatments.

Author Spotlight: A Battery of Highly Reproducible Behavioral Tests to Validate an Angelman Syndrome Murine Model 

This manuscript presents a set of highly reproducible behavioral tests to validate an Angelman syndrome mouse model.

This manuscript describes a battery of behavioral tests available to characterize Angelman syndrome (AS)-like phenotypes in an established murine model of ...

A Battery of Motor Tests in a Neonatal Mouse Model of Cerebral Palsy

As the sheer number of transgenic mice strains grow and rodent models of pediatric disease increase, there is an expanding need for a comprehensive, standardized battery of neonatal mouse motor tests. These tests can validate injury or disease models, determine treatment efficacy and/or assess motor behaviors in new transgenic strains. This paper presents a series of neonatal motor tests to evaluate general motor function, including ambulation, hindlimb foot angle, surface righting, negative geotaxis, front- and hindlimb suspension, grasping reflex, four limb grip strength and cliff aversion. Mice between the ages of post-natal day 2 to 14 can be used. In addition, these tests can be used for a wide range of neurological and neuromuscular pathologies, including cerebral palsy, hypoxic-ischemic encephalopathy, traumatic brain injury, spinal cord injury, neurodegenerative diseases, and neuromuscular disorders. These tests can also be used to determine the effects of pharmacological agents, as well as other types of therapeutic interventions. In this paper, motor deficits were evaluated in a novel neonatal mouse model of cerebral palsy that combines hypoxia, ischemia and inflammation. Forty-eight hours after injury, five tests out of the nine showed significant motor deficits: ambulation, hindlimb angle, hindlimb suspension, four limb grip strength, and grasping reflex. These tests revealed weakness in the hindlimbs, as well as fine motor skills such as grasping, which are similar to the motor deficits seen in human cerebral palsy patients.

Presented is a concise battery of mouse neonatal motor tests. Using these tests, neonatal motor deficits can be demonstrated in a variety of neonatal motor disorders. By having a standardized set of tests, results from different studies can be compared, allowing for better and accurate reporting between groups.

As the sheer number of transgenic mice strains grow and rodent models of pediatric disease increase, there is an expanding need for a comprehensive, ...

Balance and Coordination Testing

Balance and coordination are critical components involved in the control of movement. Many sensory receptors and neural processing units are required to help individuals maintain balance while performing various activities. Deficits in balance and coordination occur in patients suffering from movement disorders or due to aging. Therefore, scientists are trying to understand the pathophysiology behind these conditions. One way to do that is by using rodent models and testing them on behavioral paradigms such as the rotarod or balance beam.
This video discusses the currently known neurophysiology behind balance and coordination. Then, we go over protocols to run balance tests in rodents using the rotarod and balance beam. Finally, we'll discuss some current studies utilizing these methods to investigate aging, muscular dystrophy and Parkinson's disease.

Balance &amp; Motor Coordination: Neurophysiology, Rotarod &amp; Balance Beam Test

Balance and coordination are critical components involved in the control of movement.  Many sensory receptors and neural processing units are required to ...

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Assessing Gait by Footprint Analysis: A Method to Test Motor Coordination in Mice

Source: Sugimoto, H. and Kawakami, K. <a href="https://jove.unicartagenaproxy.elogim.com/video/59027/low-cost-protocol-footprint-analysis-hanging-box-test-for-mice">Low-cost Protocol of Footprint Analysis and Hanging Box Test for Mice Applied the Chronic Restraint Stress.</a>&#160;J. Vis. Exp. (2019).
In this video, we describe the footprint testing procedure and related analysis to assess gait and motor coordination in mice.

Source: Sugimoto, H. and Kawakami, K. Low-cost Protocol of Footprint Analysis and Hanging Box Test for Mice Applied the Chronic Restraint Stress.&#160;J. ...