Name: assessing dominant-submissive behavior in adult rats following traumatic brain injury
Uploaded: 2022-12-16T13:35:00
Description: Watch this Scientific Journal Video about Assessing Dominant-Submissive Behavior in Adult Rats Following Traumatic Brain Injury at JoVE.com

The work done are part of Dmitry Frank&#39;s PhD thesis.

The experiments were approved by the Animal Care Committee of the Ben-Gurion University of the Negev.The experiments were performed following the recommendations of the Declarations of Helsinki and Tokyo and the Guidelines for the Care and Use of Laboratory Animals of the European Community. Adult male Sprague-Dawley rats, weighing 300-350 g, were used in the present study. The animals were housed at a room temperature of 22 &#176;C &#177; 1 &#176;C and a humidity of 40%-60% with light-dark cycles.
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<ol>
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D., Tovey, C., Spangler-Martin, D., Manfredonia, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Individual+differences+versus+social+dynamics+in+the+formation+of+animal+dominance+hierarchies.">Individual differences versus social dynamics in the formation of animal dominance hierarchies.</a> Proceedings of the National Academy of Sciences of the United States of America. 99 (8), 5744-5749 (2002).</li><li>Vonk, J., Shackelford, T. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Encyclopedia of Animal Cognition and Behavior. , (2019).</li><li>De Vries, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=An+improved+test+of+linearity+in+dominance+hierarchies+containing+unknown+or+tied+relationships.">An improved test of linearity in dominance hierarchies containing unknown or tied relationships.</a> Animal Behaviour. 50 (5), 1375-1389 (1995).</li><li>Appleby, M. 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+probability+of+linearity+in+hierarchies.">The probability of linearity in hierarchies.</a> Animal Behaviour. 31 (2), 600-608 (1983).</li><li>Drent, P. J., Oers, K. v., Noordwijk, A. J. v. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Realized+heritability+of+personalities+in+the+great+tit+(Parus+major).">Realized heritability of personalities in the great tit (Parus major).</a> Proceedings of the Royal Society of London. Series B: Biological Sciences. 270 (1510), 45-51 (2003).</li><li>Sapolsky, R. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Endocrinology+alfresco:+psychoendocrine+studies+of+wild+baboons.">Endocrinology alfresco: psychoendocrine studies of wild baboons.</a> Recent Progress in Hormone Research. 48, 437-468 (1993).</li><li>Shively, C. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Social+subordination+stress,+behavior,+and+central+monoaminergic+function+in+female+cynomolgus+monkeys.">Social subordination stress, behavior, and central monoaminergic function in female cynomolgus monkeys.</a> Biological Psychiatry. 44 (9), 882-891 (1998).</li><li>Shively, C. A., Grant, K. A., Ehrenkaufer, R. L., Mach, R. H., Nader, M. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Social+stress,+depression,+and+brain+dopamine+in+female+cynomolgus+monkeys.">Social stress, depression, and brain dopamine in female cynomolgus monkeys.</a> Annals of the New York Academy of Sciences. 807, 574-577 (1997).</li><li>Tse, W. S., Bond, A. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Difference+in+serotonergic+and+noradrenergic+regulation+of+human+social+behaviours.">Difference in serotonergic and noradrenergic regulation of human social behaviours.</a> Psychopharmacology. 159 (2), 216-221 (2002).</li><li>Malatynska, E., Knapp, R. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dominant-submissive+behavior+as+models+of+mania+and+depression.">Dominant-submissive behavior as models of mania and depression.</a> Neuroscience &amp; Biobehavioral Reviews. 29 (4-5), 715-737 (2005).</li><li>Malatynska, 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=Reduction+of+submissive+behavior+in+rats:+A+test+for+antidepressant+drug+activity.">Reduction of submissive behavior in rats: A test for antidepressant drug activity.</a> Pharmacology. 64 (1), 8-17 (2002).</li><li>Frank, D., 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+life+stress+induces+submissive+behavior+in+adult+rats.">Early life stress induces submissive behavior in adult rats.</a> Behavioural Brain Research. 372, 112025 (2019).</li><li>Knapp, R. 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=Antidepressant+activity+of+memory-enhancing+drugs+in+the+reduction+of+submissive+behavior+model.">Antidepressant activity of memory-enhancing drugs in the reduction of submissive behavior model.</a> European Journal of Pharmacology. 440 (1), 27-35 (2002).</li><li>Malaty&#324;ska, E., Kostowski, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+effect+of+antidepressant+drugs+on+dominance+behavior+in+rats+competing+for+food.">The effect of antidepressant drugs on dominance behavior in rats competing for food.</a> Polish Journal of Pharmacology and Pharmacy. 36 (5), 531-540 (1984).</li><li>Kabadi, S. V., Hilton, G. D., Stoica, B. A., Zapple, D. N., Faden, A. I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Fluid-percussion-induced+traumatic+brain+injury+model+in+rats.">Fluid-percussion-induced traumatic brain injury model in rats.</a> Nature Protocols. 5 (9), 1552-1563 (2010).</li><li>Boyko, M., 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=Traumatic+brain+injury-induced+submissive+behavior+in+rats:+Link+to+depression+and+anxiety.">Traumatic brain injury-induced submissive behavior in rats: Link to depression and anxiety.</a> Translational Psychiatry. 12 (1), 239 (2022).</li><li>Jones, N. C., 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=Experimental+traumatic+brain+injury+induces+a+pervasive+hyperanxious+phenotype+in+rats.">Experimental traumatic brain injury induces a pervasive hyperanxious phenotype in rats.</a> Journal of Neurotrauma. 25 (11), 1367-1374 (2008).</li><li>Frank, D., 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+novel+histological+technique+to+assess+severity+of+traumatic+brain+injury+in+rodents:+Comparisons+to+neuroimaging+and+neurological+outcomes.">A novel histological technique to assess severity of traumatic brain injury in rodents: Comparisons to neuroimaging and neurological outcomes.</a> Frontiers in Neuroscience. 15, 733115 (2021).</li><li>Frank, D., 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+metric+test+for+assessing+spatial+working+memory+in+adult+rats+following+traumatic+brain+injury.">A metric test for assessing spatial working memory in adult rats following traumatic brain injury.</a> Journal of Visualized Experiments. (171), e62291 (2021).</li><li>Frank, D., 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=Induction+of+diffuse+axonal+brain+injury+in+rats+based+on+rotational+acceleration.">Induction of diffuse axonal brain injury in rats based on rotational acceleration.</a> Journal of Visualized Experiments. (159), e61198 (2020).</li><li>Zlotnik, 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=&#946;2+adrenergic-mediated+reduction+of+blood+glutamate+levels+and+improved+neurological+outcome+after+traumatic+brain+injury+in+rats.">&#946;2 adrenergic-mediated reduction of blood glutamate levels and improved neurological outcome after traumatic brain injury in rats.</a> Journal of Neurosurgical Anesthesiology. 24 (1), 30-38 (2012).</li><li>Frank, D., 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+novel+histological+technique+to+assess+severity+of+traumatic+brain+injury+in+rodents:+Comparisons+to+neuroimaging+and+neurological+outcomes.">A novel histological technique to assess severity of traumatic brain injury in rodents: Comparisons to neuroimaging and neurological outcomes.</a> Frontiers in Neuroscience. 15, 733115 (2021).</li><li>Marinkovic, I., 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=Prognosis+after+mild+traumatic+brain+injury:+Influence+of+psychiatric+disorders.">Prognosis after mild traumatic brain injury: Influence of psychiatric disorders.</a> Brain Sciences. 10 (12), 916 (2020).</li><li>Robert, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Traumatic+brain+injury+and+mood+disorders.">Traumatic brain injury and mood disorders.</a> Mental Health Clinician. 10 (6), 335-345 (2020).</li><li>Sabaz, M., 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=Prevalence,+comorbidities,+and+correlates+of+challenging+behavior+among+community-dwelling+adults+with+severe+traumatic+brain+injury:+A+multicenter+study.">Prevalence, comorbidities, and correlates of challenging behavior among community-dwelling adults with severe traumatic brain injury: A multicenter study.</a> The Journal of Head Trauma Rehabilitation. 29 (2), 19-30 (2014).</li><li>Aaronson, A., Lloyd, R. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Aggression+after+traumatic+brain+injury:+A+review+of+the+current+literature.">Aggression after traumatic brain injury: A review of the current literature.</a> Psychiatric Annals. 45 (8), 422-426 (2015).</li><li>Koolhaas, J. M., 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=The+resident-intruder+paradigm:+A+standardized+test+for+aggression,+violence+and+social+stress.">The resident-intruder paradigm: A standardized test for aggression, violence and social stress.</a> Journal of Visualized Experiments. (77), e4367 (2013).</li><li>Bhatnagar, S., Vining, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Facilitation+of+hypothalamic-pituitary-adrenal+responses+to+novel+stress+following+repeated+social+stress+using+the+resident/intruder+paradigm.">Facilitation of hypothalamic-pituitary-adrenal responses to novel stress following repeated social stress using the resident/intruder paradigm.</a> Hormones and Behavior. 43 (1), 158-165 (2003).</li><li>Boyko, M., 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=The+effect+of+depressive-like+behavior+and+antidepressant+therapy+on+social+behavior+and+hierarchy+in+rats.">The effect of depressive-like behavior and antidepressant therapy on social behavior and hierarchy in rats.</a> Behavioural Brain Research. 370, 111953 (2019).</li><li>Gruenbaum, B. F., 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+complex+diving-for-food+Task+to+investigate+social+organization+and+interactions+in+rats.">A complex diving-for-food Task to investigate social organization and interactions in rats.</a> Journal of Visualized Experiments. (171), e61763 (2021).</li><li>Grasmuck, V., Desor, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Behavioural+differentiation+of+rats+confronted+to+a+complex+diving-for-food+situation.">Behavioural differentiation of rats confronted to a complex diving-for-food situation.</a> Behavioural Processes. 58 (1-2), 67-77 (2002).</li><li>Pinhasov, A., Crooke, J., Rosenthal, D., Brenneman, D., Malatynska, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Reduction+of+Submissive+Behavior+Model+for+antidepressant+drug+activity+testing:+Study+using+a+video-tracking+system.">Reduction of Submissive Behavior Model for antidepressant drug activity testing: Study using a video-tracking system.</a> Behavioural Pharmacology. 16 (8), 657-664 (2005).</li><li>Nesher, 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=Differential+responses+to+distinct+psychotropic+agents+of+selectively+bred+dominant+and+submissive+animals.">Differential responses to distinct psychotropic agents of selectively bred dominant and submissive animals.</a> Behavioural Brain Research. 236 (1), 225-235 (2013).</li></ol>

Clinical studies indicate that brain injury may increase the risk of psychiatric disorders26,27. Moreover, TBI affects the development of social behavior28,29. In this protocol, the TBI model had an effect on the presentation of dominant-submissive behavior. Dominant-submissive behavior manifested itself in terms of time spent on the feeder and who came first to the feeder.
In ...

Intraspecies competition occurs when members of the same species compete for a limited resource at the same time1. In contrast, interspecies competition occurs between members of two different species2. Intraspecies competition is divided into two types, including interference (adapted) and exploitation (contest), and arises depending on the type of resource in contention, such as food and territory3.
The existence of social hierarchies is impossible without dominant-submissive relationships (DSRs). Dominance presents as &#34;winning&#34; and subordination as &#34;losing&#34; within pairs of animals4. However, DSRs appear not only in pairs but also in groups of three or more. In 1922, Thorleif Schjelderup-Ebbe described the dominance hierarchy in domestic chickens. The principal distinguishing signs between the dominant and subordinate animals were time spent at the feeder and aggressive behavior. The dominance hierarchy is divided into two forms: linear and nonlinear5. Linear dominance involves two groups, A and B. In this paradigm of transitive relationships6, group A dominates group B, or group B dominates group A. Nonlinear dominance occurs when there is at least one circular relationship: A dominates B, B dominates C, and C dominates A7.
Models for assessing dominant-submissive behavior exist for different species, including rodents, birds8, non-human primates9,10,11, and humans12. The dominant-submissive method is well represented in the literature and has been applied as a model to assess mania and depression13, as well as antidepressant drug activity14. This model has been used to investigate early life stress after maternal separation in adult rats15. The DSR paradigms can be divided into three models: the reduction of dominant behavior model13,16, the reduction of submissive behavior model14, and the clonidine-reversal of dominance model17.
This study demonstrates an investigation of DSR through tasks based on food competition. The advantages of this method are its easy reproducibility and the ability to observe and accurately analyze dominant-submissive behavior. In addition, the dominant-submissive behavioral task relies on food rather than territory, unlike comparable behavioral tasks, which makes this behavioral task lower cost and simpler and researchers do not need to undergo complicated training to perform the task and process the data.
The overall goal of the current study is to demonstrate the development of DSR after traumatic brain injury (TBI). TBI is associated with social impairments, depression, and anxiety. The model of inducing TBI is a simple and effective standard model that involves inducing traumatic brain injury with a fluid percussion device18,19.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>2% chlorhexidine in 70% alcohol solution</td>
			<td>SIGMA - ALDRICH</td>
			<td>500 cc</td>
			<td>For general antisepsis of the skin in the operatory field</td>
		</tr>
		<tr>
			<td>4 boards of different thicknesses (1.5 cm, 2.5 cm, 5 cm and 8.5 cm)</td>
			<td>This is to evaluate neurological defect</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>4-0 Nylon suture</td>
			<td>4-00</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Bottles</td>
			<td>Techniplast</td>
			<td>ACBT0262SU</td>
			<td></td>
		</tr>
		<tr>
			<td>Bupivacaine 0.1 %</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Diamond Hole Saw Drill 3 mm diameter</td>
			<td>Glass Hole Saw Kit</td>
			<td></td>
			<td>Optional.</td>
		</tr>
		<tr>
			<td>Digital Weighing Scale</td>
			<td>SIGMA - ALDRICH</td>
			<td>Rs 4,000</td>
			<td></td>
		</tr>
		<tr>
			<td>Dissecting scissors</td>
			<td>SIGMA - ALDRICH</td>
			<td>Z265969</td>
			<td></td>
		</tr>
		<tr>
			<td>Ethanol 99.9 %</td>
			<td>Pharmacy</td>
			<td></td>
			<td>5%-10% solution used to clean equipment and remove odors</td>
		</tr>
		<tr>
			<td>Fluid-percussion device</td>
			<td>custom-made at the university workshop</td>
			<td></td>
			<td>No specific brand is recommended.</td>
		</tr>
		<tr>
			<td>Gauze Sponges</td>
			<td>Fisher</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Gloves (thin laboratory gloves)</td>
			<td></td>
			<td></td>
			<td>Optional.</td>
		</tr>
		<tr>
			<td>Heater with thermometer</td>
			<td>Heatingpad-1</td>
			<td>Model: HEATINGPAD-1/2</td>
			<td>No specific brand is recommended.</td>
		</tr>
		<tr>
			<td>Horizon-XL</td>
			<td>Mennen Medical Ltd</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Isofluran, USP 100%</td>
			<td>Piramamal Critical Care, Inc</td>
			<td>NDC 66794-017</td>
			<td>Anesthetic liquid for inhalation</td>
		</tr>
		<tr>
			<td>Logitech Webcam Software</td>
			<td>Logitech</td>
			<td>2.51</td>
			<td>Software for video camera</td>
		</tr>
		<tr>
			<td>Operating forceps</td>
			<td>SIGMA - ALDRICH</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Operating Scissors</td>
			<td>SIGMA - ALDRICH</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>PC Computer for USV recording and data analyses</td>
			<td>Intel</td>
			<td></td>
			<td>Intel core i5-6500 CPU @ 3.2GHz, 16 GB RAM, 64-bit operating system</td>
		</tr>
		<tr>
			<td>Plexiglass boxes linked by a narrow passage</td>
			<td></td>
			<td></td>
			<td>Two transparent 30 cm &#215; 20 cm &#215; 20 cm plexiglass boxes linked by a narrow 15 cm &#215; 15 cm &#215; 60 cm passage</td>
		</tr>
		<tr>
			<td>Purina Chow</td>
			<td>Purina</td>
			<td>5001</td>
			<td>Rodent laboratory chow given to rats,&#160; is a lifecycle nutrition that has been used in biomedical research</td>
		</tr>
		<tr>
			<td>Rat cages (rat home cage or another enclosure)</td>
			<td>Techniplast</td>
			<td>2000P</td>
			<td>No specific brand is recommended</td>
		</tr>
		<tr>
			<td>Scalpel blades 11</td>
			<td>SIGMA - ALDRICH</td>
			<td>S2771</td>
			<td></td>
		</tr>
		<tr>
			<td>SPSS</td>
			<td>SPSS Inc., Chicago, IL, USA</td>
			<td>A 20 package</td>
			<td></td>
		</tr>
		<tr>
			<td>Stereotaxic Instrument</td>
			<td>custom-made at the university workshop</td>
			<td></td>
			<td>No specific brand is recommended</td>
		</tr>
		<tr>
			<td>Timing device</td>
			<td>Interval Timer:Timing for recording USV&#39;s</td>
			<td></td>
			<td>Optional. Any timer will do, although it is convenient to use an interval timer if you are tickling multiple rats</td>
		</tr>
		<tr>
			<td>Video camera</td>
			<td>Logitech</td>
			<td>C920 HD PRO WEBCAM</td>
			<td>Digital video camera for high definition recording of rat behavior under dominant submissive test</td>
		</tr>
	</tbody>
</table>

assessing dominant-submissive behavior in adult rats following traumatic brain injury

Competition over resources such as food, territory, and mates significantly influences relationships within animal species and is mediated through social hierarchies that are often based on dominant-submissive relationships. The dominant-submissive relationship is a normal behavioral pattern among the individuals of a species. Traumatic brain injury is a frequent cause of social interaction impairment and the reorganization of dominant-submissive relationships in animal pairs. This protocol describes submissive behavior in adult male Sprague-Dawley rats after the induction of traumatic brain injury using a fluid-percussion model compared to naive rats through a series of dominant-submissive tests performed between 29 days and 33 days after induction. The dominant-submissive behavior test shows how brain injury can induce submissive behavior in animals competing for food. After traumatic brain injury, the rodents were more submissive, as indicated by them spending less time at the feeder and being less likely to arrive first at the trough compared to the control animals. According to this protocol, submissive behavior develops after traumatic brain injury in adult male rats.

Neurological severity score assessment 
Neurological deficits were assessed in male rats after TBI using the NSS. The rats were divided into two groups: one TBI group and one control group. The control group was subjected to sham surgery. The NSS allowed for the assessment of motor function and behavior alteration by a points system22,23; a score of 24 indicated a severe neurological dysfunction, and a score of 0 represented intact neurologi...

Watch this Scientific Journal Video about Assessing Dominant-Submissive Behavior in Adult Rats Following Traumatic Brain Injury at JoVE.com

Assessing Dominant-Submissive Behavior in Adult Rats Following Traumatic Brain Injury

The present protocol describes a rat model of fluid percussion-induced traumatic brain injury followed by a series of behavioral tests to understand the development of dominant and submissive behavior. Using this model of traumatic brain injury in conjunction with specific behavioral tests enables the study of social impairments following brain injury.

Competition over resources such as food, territory, and mates significantly influences relationships within animal species and is mediated through social ...

In this study, we would like to demonstrate an investigation of dominant-submissive relationship through tests based on food competition. The overall goal of the current study is to show the development of dominant-submissive relationship after traumatic brain injury. The advantages of this method are its easy reproducibility and the ability to observe and accurately analyze dominant-submissive behavior.The dominant-submissive method is well represented in the literature and has been applied as a model to assess mania and depression. This model has been used to investigate early life stress after maternal separation in adult rats as well. To begin, select 30 adult males randomly.Divide them into traumatic brain injury and sham groups. Provide chow and water ad libitum. Before the injury, perform baseline assessments of the neurological severity score in both groups of rats as described in the manuscript.After anesthetizing the rats, check for the immobilization of the rat by testing for lack of movement or pedal reflex in response to a stimulant. Perform craniotomy four millimeter posterior and four millimeter lateral of bregma. Using a fluid percussion device, induce traumatic brain injury over 21 to 23 milliseconds through the three-way stopcock.Perform craniotomy on the group of sham operated rats and infiltrate 0.1%bupivacaine before closing the wound. One week before the test, randomly divide the rats into cages. Use an apparatus made from two transparent acrylic glass boxes connected by a slender tunnel.Next, fill a feeder with sweetened milk and place it in the tunnel center. Place each rat in the apparatus for 15 minutes for habitation on the first two days. Randomly select one rat from the control group and one from the traumatic brain injury group and set them at equal distances from the feeder, allowing them to explore for five minutes.Allow the rats access to water ad libitum. Position a camera. Install the recommended computer software to capture, save, and process the data, and record the video while the rats are in the arena.The neurological severity score showed that 48 hours after surgery, the neurological deficits were sufficiently greater for the traumatic brain injury rats compared to the sham operated rats. However, 28 days after surgery, the differences between the traumatic brain injury and sham operated groups were insignificant. Time spent at the feeder was significantly lower for the traumatic brain injury rats compared to the sham operated rats.Fewer traumatic brain injury than sham operated rats came first to the feeder. The most important things are cleaning the apparatus before every session and performing the test in a room with proper air circulation. This technique paves the way for researchers to explore dominant-submissive behavioral in rats after brain injury.

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Research

JoVE Journal

Behavior

Use of an Eight-arm Radial Water Maze to Assess Working and Reference Memory Following Neonatal Brain Injury

Working and reference memory are commonly assessed using the land based radial arm maze.&#160;However, this paradigm requires pretraining, food deprivation, and may introduce scent cue confounds. The eight-arm radial water maze is designed to evaluate reference and working memory performance simultaneously by requiring subjects to use extra-maze cues to locate escape platforms and remedies the limitations observed in land based radial arm maze designs. Specifically, subjects are required to avoid the arms previously used for escape during each testing day (working memory) as well as avoid the fixed arms, which never contain escape platforms (reference memory). Re-entries into arms that have already been used for escape during a testing session (and thus the escape platform has been removed) and re-entries into reference memory arms are indicative of working memory deficits. Alternatively, first entries into reference memory arms are indicative of reference memory deficits. We used this maze to compare performance of rats with neonatal brain injury and sham controls following induction of hypoxia-ischemia and show significant deficits in both working and reference memory after eleven days of testing. This protocol could be easily modified to examine many other models of learning impairment.

The eight-arm radial water maze is designed to evaluate reference and working memory performance simultaneously by requiring subjects to use extra-maze cues to locate escape platforms and remedies the limitations observed in land based radial arm maze designs.

Working and reference memory are commonly assessed using the land based radial arm maze.&#160;However, this paradigm requires pretraining, food ...

Operant Procedures for Assessing Behavioral Flexibility in Rats

Executive functions consist of multiple high-level cognitive processes that drive rule generation and behavioral selection. An emergent property of these processes is the ability to adjust behavior in response to changes in one&#8217;s environment (i.e., behavioral flexibility). These processes are essential to normal human behavior, and may be disrupted in diverse neuropsychiatric conditions, including schizophrenia, alcoholism, depression, stroke, and Alzheimer&#8217;s disease. Understanding of the neurobiology of executive functions has been greatly advanced by the availability of animal tasks for assessing discrete components of behavioral flexibility, particularly strategy shifting and reversal learning. While several types of tasks have been developed, most are non-automated, labor intensive, and allow testing of only one animal at a time. The recent development of automated, operant-based tasks for assessing behavioral flexibility streamlines testing, standardizes stimulus presentation and data recording, and dramatically improves throughput. Here, we describe automated strategy shifting and reversal tasks, using operant chambers controlled by custom written software programs. Using these tasks, we have shown that the medial prefrontal cortex governs strategy shifting but not reversal learning in the rat, similar to the dissociation observed in humans. Moreover, animals with a neonatal hippocampal lesion, a neurodevelopmental model of schizophrenia, are selectively impaired on the strategy shifting task but not the reversal task. The strategy shifting task also allows the identification of separate types of performance errors, each of which is attributable to distinct neural substrates. The availability of these automated tasks, and the evidence supporting the dissociable contributions of separate prefrontal areas, makes them particularly well-suited assays for the investigation of basic neurobiological processes as well as drug discovery and screening in disease models.

The ability to assess executive functions such as behavioral flexibility in rats is useful for investigating the neurobiology of cognition in both intact animals and disease models. Here we describe automated tasks for assessing strategy shifting and reversal learning, which are particularly sensitive to disruptions in prefrontal cortical networks.

Executive functions consist of multiple high-level cognitive processes that drive rule generation and behavioral selection. An emergent property of these ...

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 ...

Exploring the Use of Isolated Expressions and Film Clips to Evaluate Emotion Recognition by People with Traumatic Brain Injury

The current study presented 60 people with traumatic brain injury (TBI) and 60 controls with isolated facial emotion expressions, isolated vocal emotion expressions, and multimodal (i.e., film clips) stimuli that included contextual cues. All stimuli were presented via computer. Participants were required to indicate how the person in each stimulus was feeling using a forced-choice format. Additionally, for the film clips, participants had to indicate how they felt in response to the stimulus, and the level of intensity with which they experienced that emotion.

This paper describes how to implement a battery of behavioral tasks to examine emotion recognition of isolated facial and vocal emotion expressions, and a novel task using commercial television and film clips to assess multimodal emotion recognition that includes contextual cues.

The current study presented 60 people with traumatic brain injury (TBI) and 60 controls with isolated facial emotion expressions, isolated vocal emotion ...

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 Spatial Memory Impairment in a Mouse Model of Traumatic Brain Injury Using a Radial Water Tread Maze

Despite the recent increase in use of mouse models in scientific research, researchers continue to use cognitive tasks that were originally designed and validated for rat use. The Radial Water Tread (RWT) maze test of spatial memory (designed specifically for mice and requiring no swimming) has been shown previously to successfully distinguish between controlled cortical impact-induced TBI mice and sham controls. Here, a detailed protocol for this task is presented. The RWT maze capitalizes on the natural tendency of mice to avoid open areas in favor of hugging the sides of an apparatus (thigmotaxis). The walls of the maze are lined with nine escape holes placed above the floor of the apparatus, and mice are trained to use visual cues to locate the escape hole that leads out of the maze. The maze is filled with an inch of cold water, sufficient to motivate escape but not deep enough to require that the mouse swim. The acquisition period takes only four training days, with a test of memory retention on day five and a long-term memory test on day 12. The results reported here suggest that the RWT maze is a feasible alternative to rat-validated, swimming-based cognitive tests in the assessment of spatial memory deficits in mouse models of TBI.

Here we present a protocol for a mouse-specific test of cognition that does not require swimming. This test can be used to successfully distinguish controlled cortical impact-induced traumatic brain injury mice from sham controls.

Despite the recent increase in use of mouse models in scientific research, researchers continue to use cognitive tasks that were originally designed and ...

Detecting Behavioral Deficits in Rats After Traumatic Brain Injury

With the increasing incidence of traumatic brain injury (TBI) in both civilian and military populations, TBI is now considered a chronic disease; however, few studies have investigated the long-term effects of injury in rodent models of TBI. Shown here are behavioral measures that are well-established in TBI research for times early after injury, such as two weeks, until two months. Some of these methods have previously been used at later times after injury, up to one year, but by very few laboratories. The methods demonstrated here are a short neurological assessment to test reflexes, a Beam-Balance to test balance, a Beam-Walk to test balance and motor coordination, and a working memory version of the Morris water maze that can be sensitive to deficits in reference memory. Male rats were handled and pre-trained to neurological, balance, and motor coordination tests prior to receiving parasagittal fluid percussion injury (FPI) or sham injury. Rats can be tested on the short neurological assessment (neuroscore), the beam-balance, and the Beam-Walk multiple times, while testing on the water maze can only be done once. This difference is because rats can remember the task, thus confounding the results if repeated testing is attempted in the same animal. When testing from one to three days after injury, significant differences are detected in all three non-cognitive tasks. However, differences in the Beam-Walk task were not detectable at later time points (after 3 months). Deficits were detected at 3 months in the Beam-Balance and at 6 months in the neuroscore. Deficits in working memory were detected out to 12 months after injury, and a deficit in a reference memory first appeared at 12 months. Thus, standard behavioral tests can be useful measures of persistent behavioral deficits after FPI.

The goal of the behavioral tests presented here is to detect functional deficits in rats after traumatic brain injury. Four specific tests are presented that detect deficits in behaviors to reflect the damage to specific brain areas at times extending to one year after injury.

With the increasing incidence of traumatic brain injury (TBI) in both civilian and military populations, TBI is now considered a chronic disease; however, ...

Using Laser Doppler Imaging and Monitoring to Analyze Spinal Cord Microcirculation in Rat

Laser Doppler flowmetry (LDF) is a noninvasive method for blood flow (BF) measurement, which makes it preferable for measuring microcirculatory alterations of the spinal cord. In this article, our goal was to use both Laser Doppler imaging and monitoring to analyze the change of BF after spinal cord injury. Both the laser Doppler image scanner and the probe/monitor were being employed to obtain each readout. The data of LDPI provided a local distribution of BF, which gave an overview of perfusion around the injury site and made it accessible for comparative analysis of BF among different locations. By intensely measuring the probing area over a period of time, a combined probe was used to simultaneously measure the BF and oxygen saturation of the spinal cord, showing overall spinal cord perfusion and oxygen supply. LDF itself has a few limitations, such as relative flux, sensitivity to movement, and biological zero signal. However, the technology has been applied in clinical and experimental study due to its simple setup and rapid measurement of BF.

Here we present a combination of laser Doppler perfusion imaging (LDPI) and laser Doppler perfusion monitoring (LDPM) to measure spinal cord local blood flows and oxygen saturation (SO2), as well as a standardized procedure for introducing spinal cord trauma on rat.

Laser Doppler flowmetry (LDF) is a noninvasive method for blood flow (BF) measurement, which makes it preferable for measuring microcirculatory ...

The Knob Supination Task: A Semi-automated Method for Assessing Forelimb Function in Rats

Tasks that accurately measure dexterity in animal models are critical to understand hand function. Current rat behavioral tasks that measure dexterity largely use video analysis of reaching or food manipulation. While these tasks are easy to implement and are robust across disease models, they are subjective and laborious for the experimenter. Automating traditional tasks or creating new automated tasks can make the tasks more efficient, objective, and quantitative. Since rats are less dexterous than primates, central nervous system (CNS) injury produces more subtle deficits in dexterity, however, supination is highly affected in rodents and crucial to hand function in primates. Therefore, we designed a semi-automated task that measures forelimb supination in rats. Rats are trained to reach and grasp a knob-shaped manipulandum and turn the manipulandum in supination to receive a reward. Rats can acquire the skill within 20 &plusmn; 5 days. While the early part of training is highly supervised, much of the training is done without direct supervision. The task reliably and reproducibly captures subtle deficits after injury and shows functional recovery that accurately reflects clinical recovery curves. Analysis of data is performed by specialized software through a graphical user interface that is designed to be intuitive. We also give solutions to common problems encountered during training, and show that minor corrections to behavior early in training produce reliable acquisition of supination. Thus, the knob supination task provides efficient and quantitative evaluation of a critical movement for dexterity in rats.

This manuscript describes a semi-automated task that quantifies supination in rats. Rats reach, grasp, and supinate a spherical manipulandum. The rat is rewarded with a pellet if the turn angle exceeds a criterion set by the user. This task increases throughput, sensitivity to injury, and objectivity compared to traditional tasks.

Tasks that accurately measure dexterity in animal models are critical to understand hand function. Current rat behavioral tasks that measure dexterity ...

A Conditioned Place Preference Protocol for Measuring Incubation of Craving in Rats

A major cause of repeated relapses is a craving for the drug. Drug craving increases progressively during the abstinence period, a phenomenon termed incubation of drug craving. Here, we describe a morphine conditioned place preference (CPP) protocol for measuring the incubation of craving in rats. In this protocol, a CPP paradigm mainly employing somatosensory cues is used to establish a long-term reward memory of morphine. A three-chamber CPP box that differs in the texture of the chamber floor is constructed. First, the animals are tested for their baseline preference to the two side chambers for three consecutive days. Then, they are injected intraperitoneally with morphine/saline and put into their non-preferred/preferred chamber for 45 min. After 6 days of conditioning, their preference to the side chambers is tested for 15 min at different time points after the last conditioning session. With this paradigm, the reward memory of morphine could last for at least 18 days. To test whether the above-mentioned protocol can model increased craving, the number of entrances into the two side chambers are counted during the abstinence period. The results show that the entrances increased, suggesting that the CPP paradigm could mimic the incubation of craving. Future studies can employ this model to study neural mechanisms underlying long-term memory and incubation of craving.

Here, a morphine conditioned place preference (CPP) protocol is described to measure incubation of craving in rats.

A major cause of repeated relapses is a craving for the drug. Drug craving increases progressively during the abstinence period, a phenomenon termed ...