This work was supported by The National Institute of Health (NIH-2RO1NS033661) and by the Alabama Agricultural Experiment Station (HATCH ALA021-1-09017).

NOTE: All procedures performed here were submitted to and approved by IACUC (Office of Research Compliance) and were conducted following NIH guidelines. Mice used in the behavioral testing paradigm were naive and not used for other tests. The C57BL/6 Wild Type and Knock Out mice used in this protocol have been described previously9 and the data presented here are from that manuscript.
1. Preparation of the Testing Room and Open Field Apparatus
<ol>
	<li>Use a multiple unit open fi...

<ol>
	<li>Hall, C. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Emotional+behavior+in+the+rat:+defecation+and+urination+as+measures+of+individual+differences+in+emotionality.">Emotional behavior in the rat: defecation and urination as measures of individual differences in emotionality.</a> J. Comp. Psychol. 18, 385-403 (1934).</li><li>Walsh, R. N., Cummins, R. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+open+field+test:+a+critical+review.">The open field test: a critical review.</a> Psychol. Bull. 83, 482-504 (1976).</li><li>Prut, L., Belzung, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+open+field+as+a+paradigm+to+measure+the+effects+of+drugs+on+anxiety-like+behaviors:+a+review.">The open field as a paradigm to measure the effects of drugs on anxiety-like behaviors: a review.</a> Eur. J. of Pharm. 463, 3-33 (2003).</li><li>Choleris, E., Thomas, A. W., Kavaliers, M., Prato, F. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+detailed+ethological+analysis+of+the+mouse+open+field+test:+effects+of+diazepam,+chlordiazpoxide+and+an+extremely+low+frequency+pulsed+magnetic+field.">A detailed ethological analysis of the mouse open field test: effects of diazepam, chlordiazpoxide and an extremely low frequency pulsed magnetic field.</a> Neurosci. Biobehav. Rev. 25, 235-260 (2001).</li><li>Ramos, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Animal+Models+of+anxiety:+do+I+need+multiple+tests.">Animal Models of anxiety: do I need multiple tests.</a> TIPS. 29, 493-498 (2008).</li><li>Archer, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Tests+for+emotionality+in+rats+and+mice:+a+review.">Tests for emotionality in rats and mice: a review.</a> Anim. Behav. 21, 205-235 (1973).</li><li>Gray, J. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Emotionality+in+male+and+female+rodents:+a+reply+to+Archer.">Emotionality in male and female rodents: a reply to Archer.</a> Brit. J. Psych. 70, 425-440 (1979).</li><li>Carola, V., D&#8217;Olimpio, F., Brunamonti, E., Mangia, F., Renzi, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Evaluation+of+the+elevated+plus-maze+and+open-field+tests+for+the+assessment+of+anxiety-related+behavior+in+inbred+mice.">Evaluation of the elevated plus-maze and open-field tests for the assessment of anxiety-related behavior in inbred mice.</a> Behav. Brain Res. 134, 49-57 (2002).</li><li>Ramesh Babu, J., Seibenhener, M. L., Peng, J., Strom, A. L., Kemppainen, R., Cox, N., Zhu, H., Wooten, M. C., Diaz-Meco, M. T., Moscat, J., Wooten, M. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Genetic+inactivation+of+p62+leads+to+accumulation+of+hyperphosphorylated+tau+and+neurodegeneration.">Genetic inactivation of p62 leads to accumulation of hyperphosphorylated tau and neurodegeneration.</a> J. Neurochem. 106, 107-120 (2008).</li><li>Simon, P., Dupuis, R., Costentin, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Thigmotaxis+as+an+index+of+anxiety+in+mice:+influence+of+dopaminergic+transmissions.">Thigmotaxis as an index of anxiety in mice: influence of dopaminergic transmissions.</a> Behav. Brain Res. 61, 59-64 (1994).</li><li>Denenberg, V. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Open-field+behavior+in+the+rat:+what+does+it+mean.">Open-field behavior in the rat: what does it mean.</a> Ann. N.Y. Acad. Sci. 159, 852-859 (1969).</li><li>Stanford, S. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+open+field+test:+reinventing+the+wheel.">The open field test: reinventing the wheel.</a> J. Psychopharm. 21, 134-135 (2007).</li><li>Ennaceur, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Tests+of+unconditional+anxiety+&#8211;+pitfalls+and+disappointments.">Tests of unconditional anxiety &#8211; pitfalls and disappointments.</a> J. Phys. Behav. 135, 55-71 (2014).</li><li>Borta, A., Schwarting, R. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Inhibitory+avoidance,+pain+reactivity,+and+plus-maze+behavior+in+Wistar+rats+with+high+versus+low+rearing+activity.">Inhibitory avoidance, pain reactivity, and plus-maze behavior in Wistar rats with high versus low rearing activity.</a> J. Phys. Behav. 84, 387-396 (2005).</li><li>Costall, B., Jones, B. J., Kelly, M. E., Naylor, R. J., Tomkins, D. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Exploration+of+mice+in+a+black+and+white+test+box:+validation+as+a+model+of+anxiety.">Exploration of mice in a black and white test box: validation as a model of anxiety.</a> Pharmacol. Biochem. Behav. 32, 777-785 (1989).</li><li>Bouwknecht, J. A., Paylor, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pitfalls+in+the+interpretation+of+genetic+and+pharmacological+effects+on+anxiety-like+behavior+in+rodents.">Pitfalls in the interpretation of genetic and pharmacological effects on anxiety-like behavior in rodents.</a> Behav. Pharm. 19, 385-402 (2008).</li><li>Webster, D. G., Baumgardner, D. J., Dewsbury, D. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Open+field+behavior+in+eight+taxa+of+muriod+rodents.">Open field behavior in eight taxa of muriod rodents.</a> Bull. Psychonom. Soc. 13, 90-92 (1979).</li><li>Wilson, R. C., Vacek, T., Lanier, D. L., Dewsbury, D. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Open+field+behavior+in+muroid+rodents.">Open field behavior in muroid rodents.</a> Behav. Biol. 17, 495-506 (1976).</li><li>Crawley, J. N. <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+transgenic+and+knockout+mice:+experimental+designs+and+evaluation+of+general+health,+sensory+functions,+motor+abilities+and+specific+behavioral+tests.">Behavioral phenotyping of transgenic and knockout mice: experimental designs and evaluation of general health, sensory functions, motor abilities and specific behavioral tests.</a> Brain Res. 835, 18-26 (1999).</li><li>Simon, P., Dupuis, R., Costentin, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Thigmotaxis+as+an+index+of+anxiety+in+mice.+influence+of+dopaminergic+transmissions.">Thigmotaxis as an index of anxiety in mice. influence of dopaminergic transmissions.</a> Behav. Brain Res. 61, 59-64 (1994).</li><li>Miller, B. H., Schultz, L. E., Gulati, A., Su, A. I., Pletcher, M. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Phenotypic+characterization+of+a+genetically+diverse+panel+of+mice+for+behavioral+despair+and+anxiety.">Phenotypic characterization of a genetically diverse panel of mice for behavioral despair and anxiety.</a> PLoS One. 5, e14458 (2010).</li><li>Tannenbaum, B., Anisman, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Impact+of+chronic+intermittent+challenges+in+stressor-susceptible+and+resilient+strains+of+mice.">Impact of chronic intermittent challenges in stressor-susceptible and resilient strains of mice.</a> Biol. Psych. 53, 292-303 (2003).</li><li>Lister, R. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ethologically-based+animal+models+of+anxiety+disorders.">Ethologically-based animal models of anxiety disorders.</a> Pharmacol. Ther. 46, 321-340 (1990).</li><li>Kulesskaya, N., Voikar, V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Assessment+of+mouse+anxiety-like+behavior+in+the+light-dark+box+and+open-field+arena:+role+of+equipment+and+procedure.">Assessment of mouse anxiety-like behavior in the light-dark box and open-field arena: role of equipment and procedure.</a> Phys. Behav. 133, 30-38 (2014).</li><li>Han, H., Du, W., Zhou, B., Zhang, W., Xu, G., Niu, R., Sun, Z. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effects+of+chronic+fluoride+exposure+on+object+recognition+memory+and+mRNA+expression+of+SNARE+complex+in+hippocampus+of+male+mice.">Effects of chronic fluoride exposure on object recognition memory and mRNA expression of SNARE complex in hippocampus of male mice.</a> Biol. Trace Elem. Res. 158, 58-64 (2014).</li><li>Barrow, P., Leconte, I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+influence+of+body+weight+on+open+field+and+swimming+maze+performance+during+the+post-weaning+period+in+the+rat.">The influence of body weight on open field and swimming maze performance during the post-weaning period in the rat.</a> Lab. Animals. 30, 22-27 (1996).</li></ol>

The authors have nothing to disclose.

The Open Field Maze is one of the most widely used platforms in animal behavioral studies. A number of important conventional and ethological parameters2,4 can be collected and analyzed during the performance of the OFM. These data allow the researcher to measure behaviors ranging from overall locomotor activity to anxiety-related emotional behaviors8. However, use of OFM is not without its shortcomings. One confounding issue is the wide range of static variables that can be manipulated during any t...

The Open Field Maze (OFM) was initially developed in 1934 as a test to measure emotionality in rodents1. It has attained the status of being one of the most widely used measures of behavior in animal psychology2. It provides an easy and fairly rapid assessment of well-defined behaviors requiring no training to the test subject and little to no specialized training for the human administering the test. These attributes have led to wide-spread use of the open field maze in research extended to other animal species such as calves, pigs, rabbits, primates, honeybees and lobsters3. Part of its popularity arises from the fact that the psychological and physiological concepts underlying the tests are generally straight-forward and well understood. For example, it has been postulated that evolutionary forces have selected for a common response in animals such that most species display anxiety-mediated fear or flight responses to specific stimuli. Rodents for example, show distinct aversions to large, brightly lit, open and unknown environments4. We can assume they have been phylogenetically conditioned to see these types of environments as dangerous. All of these features are incorporated in the open field maze and form the basis of its use in behavioral paradigm testing.
An open field maze consists of a wall-enclosed area that is of sufficient height to prevent the subject from escaping. Typical maze shapes are circular or square with an area large enough, based on the size of the subject tested, to elicit a feeling of openness in the center of the maze. A number of variables can be scored in the open field maze with most parameters involving differing types of motor activity2. Ambulation is the most common behavior studied but others such as latency or rearing can also be measured. Most often, rodent behavior is analyzed in a bare maze. However, the addition of objects, either one or many to the maze floor, adds the ability to see how the subject interacts with novel additional stimuli2. Relevant parameters when objects are presented are typically the number of approaches to an object or in some cases, preference or aversion for one object over another.
Many behavioral tests of anxiety are based on the subject animal&#8217;s body activity and locomotion5. Interpreting behavioral tests for emotionality while separating non-emotional confounding factors, such as motor activity, has been the subject of intense debate6,7. As the OFM was originally described, two measures of emotionality can be deduced, locomotor activity and fecal boli deposits or defecation1. However, these two measures have been shown in some studies to be unrelated supporting the conclusion that emotionality in rodents is multidimensional5. Regardless, discrepancies in the literature regarding these measures and emotionality or anxiety in mouse models may be attributed to differences in analysis criteria or differences in testing procedures. Studies have conclusively linked results from OFM analysis with other measures of anxiety when comparing mouse models8.

<table border="0" cellpadding="0" cellspacing="0" width="631">
	<tbody>
		<tr height="40">
			<td height="40" style="height:40px;width:160px;">Multi Unit Open Field Test</td>
			<td style="width:171px;">San Diego Instruments, Inc.</td>
			<td style="width:109px;">White 7001-0354</td>
			<td style="width:192px;">Any single or multi unit open field maze can be used</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:160px;">SMART DT Tracking Software</td>
			<td style="width:171px;">PanLab/Harvard Apparatus</td>
			<td style="width:109px;">76-0695</td>
			<td style="width:192px;">Any tracking software can be utilized with this protocol</td>
		</tr>
		<tr height="60">
			<td height="60" style="height:60px;width:160px;">Sony 990x Video Camera Recorder</td>
			<td style="width:171px;">Sony</td>
			<td style="width:109px;">CCD-TRV328</td>
			<td style="width:192px;">Any suitable video camera can be attached to computer for recording tracking profiles.</td>
		</tr>
	</tbody>
</table>

use of the open field maze to measure locomotor and anxiety-like behavior in mice

Animal models have proven to be invaluable to researchers trying to answer questions regarding the mechanisms of behavior. The Open Field Maze is one of the most commonly used platforms to measure behaviors in animal models. It is a fast and relatively easy test that provides a variety of behavioral information ranging from general ambulatory ability to data regarding the emotionality of the subject animal. As it relates to rodent models, the procedure allows the study of different strains of mice or rats both laboratory bred and wild-captured. The technique also readily lends itself to the investigation of different pharmacological compounds for anxiolytic or anxiogenic effects. Here, a protocol for use of the open field maze to describe mouse behaviors is detailed and a simple analysis of general locomotor ability and anxiety-related emotional behaviors between two strains of C57BL/6 mice is performed. Briefly, using the described protocol we show Wild Type mice exhibited significantly less anxiety related behaviors than did age-matched Knock Out mice while both strains exhibited similar ambulatory ability.

The average number of individuals per strain of mice tested in most cases is approximately 20 to generate sufficient statistical relevance. However, this number can be in the range of 8-30 depending on mouse availability. Depending on the measurement or comparisons required, it is also favorable to use age-matched subjects.
The first and arguably most important specific parameter to measure in the Open Field Maze is total ambulatory distance. While the unit of measure is irrelevant for compari...

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Use of the Open Field Maze to Measure Locomotor and Anxiety-like Behavior in Mice

A protocol is provided to use an Open Field Maze to access general locomotor activity, anxiety and emotionality in a laboratory mouse model.

Animal models have proven to be invaluable to researchers trying to answer questions regarding the mechanisms of behavior. The Open Field Maze is one of ...

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70.6K vistas.  Auburn University. A protocol is provided to use an Open Field Maze to access general locomotor activity, anxiety and emotionality in a laboratory mouse model.