Abstract
A major impediment to novel drug development has been the paucity of animal models that accurately reflect symptoms of affective disorders. In animal models, prolonged social stress has proven to be useful in understanding the molecular mechanisms underlying affective-like disorders. When considering experimental approaches for studying depression, social defeat stress, in particular, has been shown to have excellent etiological, predictive, discriminative and face validity. Described here is a protocol whereby C57BL/6J mice that are repeatedly subjected to bouts of social defeat by a larger and aggressive CD-1 mouse results in the development of a clear depressive-like syndrome, characterized by enduring deficits in social interactions. Specifically, the protocol consists of three important stages, beginning with the selection of aggressive CD-1 mice, followed by agonistic social confrontations between the CD-1 and C57BL/6J mice, and concluding with the confirmation of social avoidance in subordinate C57BL/6J mice. The automated detection of social avoidance allows a marked increase in throughput, reproducibility and quantitative analysis. This protocol is highly adaptable, but in its most common form it requires 3–4 weeks for completion.
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17 December 2014
In the version of this article initially published, there was some confusion over the interpretation of the sentence "Further, defeats should be run under constant veterinary evaluation and with full approval of all necessary institutional review boards and standards." For added clarity, the sentence was changed to read "Further, defeats should be run with full approval of all necessary institutional review boards and standards." The error has been corrected in the HTML and PDF versions of the article.
References
Greenberg, P.E. et al. The economic burden of depression in the United States: how did it change between 1990 and 2000? J. Clin. Psychiatry 64, 1465–1475 (2003).
Krishnan, V. & Nestler, E.J. Linking molecules to mood: new insight into the biology of depression. Am. J. Psychiatry 167, 1305–1320 (2008).
Krishnan, V. & Nestler, E.J. The molecular neurobiology of depression. Nature 455, 894–902 (2008).
Nestler, E.J. & Hyman, S.E. Animal models of neuropsychiatric disorders. Nat. Neurosci. 13, 1161–1169 (2010).
Li, N. et al. mTOR-dependent synapse formation underlies the rapid antidepressant effects of NMDA antagonists. Science 329, 959–964 (2010).
Yehuda, R., Flory, J.D., Southwick, S. & Charney, D.S. Developing an agenda for translational studies of resilience and vulnerability following trauma exposure. Ann. NY Acad. Sci. 1071, 379–396 (2006).
Butterweck, V., Winterhoff, H. & Herkenham, M. St John's wort, hypericin, and imipramine: a comparative analysis of mRNA levels in brain areas involved in HPA axis control following short-term and long-term administration in normal and stressed rats. Mol. Psychiatry 6, 547–564 (2001).
Rygula, R. et al. Pharmacological validation of a chronic social stress model of depression in rats: effects of reboxetine, haloperidol and diazepam. Behav. Pharmacol. 19, 183–196 (2008).
Koolhaas, J.M., De Boer, S.F., De Rutter, A.J., Meerlo, P. & Sgoifo, A. Social stress in rats and mice. Acta. Physiol. Scand. Suppl. 640, 69–72 (1997).
Kudryavtseva, N.N., Bakshtanovskaya, I.V. & Koryakina, L.A. Social model of depression in mice of C57BL/6J strain. Pharmacol. Biochem. Behav. 38, 315–320 (1991).
Miczek, K.A., Yap, J.J. & Covington, H.E. III. Social stress, therapeutics and drug abuse: preclinical models of escalated and depressed intake. Pharmacol. Ther. 120, 102–128 (2008).
Brain, P.F. Mammalian behavior and the adrenal cortex. A review. Behav. Biol. 7, 453–477 (1972).
Fuchs, E. & Flugge, G. Social stress in tree shrews: effects on physiology, brain function, and behavior of subordinate individuals. Pharmacol. Biochem. Behav. 73, 247–258 (2002).
Rygula, R., Abumaria, N., Domenici, E., Hiemke, C. & Fuchs, E. Effects of fluoxetine on behavioral deficits evoked by chronic social stress in rats. Behav. Brain. Res. 174, 188–192 (2006).
Rygula, R. et al. Citalopram counteracts depressive-like symptoms evoked by chronic social stress in rats. Behav. Pharmacol. 17, 19–29 (2006).
Rygula, R. et al. Anhedonia and motivational deficits in rats: impact of chronic social stress. Behav. Brain. Res. 162, 127–134 (2005).
Berton, O. et al. Essential role of BDNF in the mesolimbic dopamine pathway in social defeat stress. Science 311, 864–868 (2006).
Tornatzky, W. & Miczek, K.A. Long-term impairment of autonomic circadian rhythms after brief intermittent social stress. Physiol. Behav. 53, 983–993 (1993).
Covington, H.E. III & Miczek, K.A. Intense cocaine self-administration after episodic social defeat stress, but not after aggressive behavior: dissociation from corticosterone activation. Psychopharmacology (Berl) 183, 331–340 (2005).
Berton, O. & Nestler, E.J. New approaches to antidepressant drug discovery: beyond monoamines. Nat. Rev. Neurosci. 7, 137–151 (2006).
Krishnan, V. et al. Molecular adaptations underlying susceptibility and resistance to social defeat in brain reward regions. Cell 131, 391–404 (2007).
Avgustinovich, D.F., Gorbach, O.V. & Kudryavtseva, N.N. Comparative analysis of anxiety-like behavior in partition and plus-maze tests after agonistic interactions in mice. Physiol. Behav. 61, 37–43 (1997).
Lutter, M. et al. The orexigenic hormone ghrelin defends against depressive symptoms of chronic stress. Nat. Neurosci. 11, 752–753 (2008).
Chuang, J.C. et al. Chronic social defeat stress disrupts regulation of lipid synthesis. J. Lipid Res. 51, 1344–1353 (2010).
Chuang, J.C. et al. A beta3-adrenergic-leptin-melanocortin circuit regulates behavioral and metabolic changes induced by chronic stress. Biol. Psychiatry 67, 1075–1082 (2010).
Lutter, M. et al. Orexin signaling mediates the antidepressant-like effect of calorie restriction. J. Neurosci. 28, 3071–3075 (2008).
Tsankova, N.M. et al. Sustained hippocampal chromatin regulation in a mouse model of depression and antidepressant action. Nat. Neurosci. 9, 519–525 (2006).
Li, N. et al. Glutamate N-methyl-D-aspartate receptor antagonists rapidly reverse behavioral and synaptic deficits caused by chronic stress exposure. Biol. Psychiatry 69, 754–761 (2011).
Covington, H.E. III et al. Antidepressant actions of histone deacetylase inhibitors. J. Neurosci. 29, 11451–11460 (2009).
Wilkinson, M.B. et al. Imipramine treatment and resiliency exhibit similar chromatin regulation in the mouse nucleus accumbens in depression models. J. Neurosci. 29, 7820–7832 (2009).
Vialou, V. et al. [Delta]FosB in brain reward circuits mediates resilience to stress and antidepressant responses. Nat. Neurosci. 13, 745–752 (2010).
Christoffel, D.J. et al. IkappaB kinase regulates social defeat stress-induced synaptic and behavioral plasticity. J. Neurosci. 31, 314–321 (2011).
Johren, O., Flugge, G. & Fuchs, E. Regulation of hippocampal glucocorticoid receptor gene expression by psychosocial conflict. Ann. NY Acad. Sci. 746, 429–430 (1994).
Blanchard, D.C. et al. Visible burrow system as a model of chronic social stress: behavioral and neuroendocrine correlates. Psychoneuroendocrinology 20, 117–134 (1995).
Koolhaas, J.M., Everts, H., de Ruiter, A.J., de Boer, S.F. & Bohus, B. Coping with stress in rats and mice: differential peptidergic modulation of the amygdala-lateral septum complex. Prog. Brain Res. 119, 437–448 (1998).
Vivian, J.A. & Miczek, K.A. Interactions between social stress and morphine in the periaqueductal gray: effects on affective vocal and reflexive pain responses in rats. Psychopharmacology (Berl) 146, 153–161 (1999).
Nestler, E.J. et al. Neurobiology of depression. Neuron 34, 13–25 (2002).
Huhman, K.L. Social conflict models: can they inform us about human psychopathology? Horm. Behav. 50, 640–646 (2006).
Dadomo, H. et al. Vulnerability to chronic subordination stress-induced depression-like disorders in adult 129SvEv male mice. Prog. Neuropsychopharmacol. Biol. Psychiatry (17 November 2010).
Razzoli, M. et al. Strain-specific outcomes of repeated social defeat and chronic fluoxetine treatment in the mouse. Pharmacol. Biochem. Behav. 97, 566–576 (2010).
Razzoli, M., Carboni, L., Andreoli, M., Ballottari, A. & Arban, R. Different susceptibility to social defeat stress of BALB/c and C57BL6/J mice. Behav. Brain Res. 216, 100–108 (2010).
Gimsa, U., Kanitz, E., Otten, W. & Ibrahim, S.M. Behavior and stress reactivity in mouse strains with mitochondrial DNA variations. Ann. NY Acad. Sci. 1153, 131–138 (2009).
Haenisch, B., Bilkei-Gorzo, A., Caron, M.G. & Bonisch, H. Knockout of the norepinephrine transporter and pharmacologically diverse antidepressants prevent behavioral and brain neurotrophin alterations in two chronic stress models of depression. J. Neurochem. 111, 403–416 (2009).
Covington, H.E. III et al. Antidepressant effect of optogenetic stimulation of the medial prefrontal cortex. J. Neurosci. 30, 16082–16090 (2010).
Acknowledgements
We thank D. Christoffel for his helpful review of this manuscript. We also acknowledge the intrepid efforts of the Mount Sinai School of Medicine animal facility personnel. This research was supported by US National Institute of Mental Health grant 1R01MH090264-01A1.
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S.A.G., H.E.C., O.B. and S.J.R. contributed to study design, data collection, analysis and writing.
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Golden, S., Covington, H., Berton, O. et al. A standardized protocol for repeated social defeat stress in mice. Nat Protoc 6, 1183–1191 (2011). https://doi.org/10.1038/nprot.2011.361
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DOI: https://doi.org/10.1038/nprot.2011.361
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Natalia Kudryavtseva
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Standardized Protocol for Repeated Social Defeat Stress vs. Sensory Contact Model: similarities and differences, strengths and weaknesses
Kudryavtseva N.N.
Neurogenetics of Social Behavior Sector, Institute of Cytology and Genetics SD RAS, Novosibirsk, Russia
Of all the models used by behavioral neuroscience the biosocial models are gaining in popularity among the scientists who study the consequences of chronic social conflicts and social stress in animals. One of such models presented as the Standardized Protocol of Repeated Social Defeat Stress(SPRSDS)<sup>1</sup> has been widely applied to produce important results on the molecular mechanisms of the social defeat stress<sup>2</sup>. SPRSDS is a modified version of the Sensory Contact Model<sup>3</sup> (SCM), which has been used by different laboratories in its original or modified forms for elucidating implications of chronic social conflicts. The basic design and justification of the SCM different stages are described in detail in the latest publication “Sensory contact model: Protocol, control, applications” <sup>4</sup>. Thus, it is now possible to compare similarities, differences and, as a consequence, strengths and weaknesses of these two behavioral paradigms.
SPRSDS and SCM designs: principal similarities
· An experimental cage contains a perforated transparent Plexiglas partition (screen, divider) dividing the cage into two compartments;
· During the experiments defeated and aggressor mice are housed in the neighboring compartments of common cages;
· Defeated mice are daily transferred to novel cages with unfamiliar aggressors in the neighboring compartments.
SPRSDS and SCM designs: principal differences
Size and design of experimental cage:
SPRSDS: Rectangular hamster cages 26.7 cm (w) × 48.3 cm (d) × 15.2 cm (h) divided lengthwise into two equal compartments by a perforated Plexiglas divider which physically separates the mice after fighting sessions;
SCM: Special cages 14 cm х 28 cmх 10 cm divided crosswise by a perforated Plexiglas partition into two equal compartments. The partition is removed daily to allow agonistic interactions on common territory.
SPRSDS: Preliminarily screened, heaver and older males of CD-1 strain are used as aggressors. C57BL/6J mice are used as defeaters.
SCM: Males of the same strain, similar age and weight are used as aggressors and defeaters: In the first agonistic interactions the winners and defeaters become obvious (for detail see <sup> 4</sup>).
SPRSDS: During agonistic interaction test defeated males are placed in aggressors’ compartments.
SCM: The partition is removed from the cage to start agonistic interaction between mice on common territories.
SPRSDS: Some critical steps used in the SCM are lacking: 2-3 days of sensory contact and 5 minute of activation before agonistic interactions etc. (for detail see<sup> 4</sup>).
Different social motivations in behavior of defeated mice:
SPRSDS: Resident-intruder behavior, the intruder (defeated mice) demonstrates defense behavior on resident’s (aggressor’s) territory;
SCM:Loss of social status for the defeaters in the first agonistic interactions.
Different controls:
SPRSDS: Control mice are pair housed in defeat boxes to each side of the perforated divider. All control mice are rotated on a daily basis in a manner similar to that of the mice undergoing defeat, but they are never actually allowed physical contact with their cage mates.
Thus, control mice are subjected to all stress procedures excluding physical contact with aggressors.
SCM: 5-7 days of individual housing. The problem of the control in the SCM and rationality of this control which was confirmed in special experiments are discussed in detail in <sup>4</sup>.
Key etiologic factor:
SPRSDS: Physical attacks of aggressor;
SCM: Chronic psychoemotional stress: expectation of adverse events, waiting for attacks from aggressive male.
Strengths of SPRSDS
The model can be used in limited situations (lack of special cages) for the study of prolonged social avoidance induced by 10 days chronic social defeat stress under the lack of aggressive behavior in male mice of the same strain.
Weaknesses of SPRSDS
Aggressor mice are big and old mice of CD-1 strain which have to be preliminarily screened. Only 50% of CD-1 mice can be used in the experimentation as aggressors; 30-40% of the mice of inbred C57BL/6J strain subjected to social defeat stress are not susceptible to stress. Invalid control – stressed mice.
On the molecular level: the absence of changes in the expression of specific genes whose products are involved in the brain mechanisms of anxiety or depression, for example, serotonergic genes <sup>2</sup>.
Strengths of SCM
90-100% of the mice of C57BL/6J strain demonstrate aggression or active defense in the first agonistic interaction. Thereafter 90-100% of the victorious mice demonstrate aggression daily. The partition test, which measures behavioral reaction (communicativeness) to familiar or unfamiliar partners in the neighboring compartment, is very informative for testing the level of avoidance behavior in defeaters and expression of aggressivemotivation in the victorious mice<sup>5</sup>. The results of different laboratories prove a good reproducibility of the method<sup>6</sup>. A wide range of psychoemotional disorders (not only prolonged social avoidance) can be formed in male mice under repeated agonistic interactions on the basis of different mouse strains. Therefore, it is possible to carry out pharmacological screening of different psychotropic drugs under simulated clinical conditions<sup>7</sup> and to study the dynamic mechanisms of the evolving psychoemotional disorders from norm to severe pathology. Stability of the model has been demonstrated over two decades of its application.
Weaknesses of SCM
The innovative behavioral approach to the modeling of psychoemotional disorders, including a novel approach to control choice (healthy state and social norm vs. conventional control) and the system of comparison (norm vs. psychopathology), is difficult for assimilation by the newcomers and reviewers. The lack of necessary knowledge of the behavioral and neurophysiologic processes working in the SCM and insufficient experience in their modeling might lead to false interpretation of the results. A study of a forming psychopathology requires prolonged experiments. For example, the experiment on the induction of mixed anxiety/depression state and collection of all the evidence may take 4-6 weeks. For each new mouse strain and for different laboratory conditions the experimenter may need to introduce some modifications into the preliminary or testing procedures.
However, for the experimenters who decide to use the original version of SCM its weaknesses will be outweighed by its wide applicability in the modeling and studies of different psychoemotional disorders (anxious depression, catalepsy, social avoidance, pathological aggression, generalized anxiety, hyperactivity, disturbances in cognition and motivated behaviors, anhedonia, addictive state etc.) accompanied by somatic changes (reduced gonad function, psychogenic immune deficiency, enhanced metastasis etc.) as well as in the studies of the effects of psychotropic drugs under simulated clinical conditions.
References
1. Golden, S.A., Covington III, H.E., Berton, O., Russo, S.J. A standardized protocol for repeated social defeat stress in mice. Nature Prot. 6, 8,1183-1991 (2011).
2. Berton, O. et al. Essential role of BDNF in the mesolimbic dopamine pathway in social defeat stress. Science 311, 864�(2006).
3. Kudryavtseva, N.N. The sensory contact model for the study of aggressive and submissive behaviors in male mice. Aggres. Behav. 17 (5), 285-291(1991).
4. Kudryavtseva, N.N. Sensory contact model: Protocol, control, applications. In Horizons in Neuroscience Research, NOVA Science Publishers Inc., Editors: Andres Costa and Eugenio Villalba, Vol. 3, Chapter 4, 81-100, New York (2011).https://www.novapublishers.com/cata...
5. Kudriavtseva, N.N. Use of the "partition" test in behavioral and pharmacological experiments. Neurosci. Behav. Physiol. 33 (5), 461-471 (2003).
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6.Bartolomucci, A., Fuchs, E., Koolhaas, J.M., Ohl, F. Acute and Chronic Social Defeat: Stress Protocols and Behavioral Testing Mood and Anxiety Related Phenotypes in Mice, Neuromethods, Chapter 14, Ed. Todd D. Gould, Humana Press, 2009, 261-275.
7.Kudryavtseva, N.N. et al. An experimental approach for the study of psychotropic drug effects under simulated clinical conditions. Curr. Drug Metab. 9, 4, 352-360 (2008).
Bronwen Dekker
The comment above from Natalia Kudryavtseva compares two approaches:
(1) A standardized protocol for repeated social defeat stress in mice (the subject of this Nature Protocol)
(2) The sensory contact model
In addition to the refereces included in the comment itself, I would like to highlight that an open-access protocol for the Sensory contact model can be found at Nature Precedings.
Sensory Contact Model: Protocol, Control, Applications