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Behavioral methods to study anxiety in rodents
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Stress is a precipitating factor for anxiety-related disorders, which are among the leading forms of psychiatric illness and impairment in the modern world. Rodent-based behavioral tests and models are widely used to understand the mechanisms by which stress triggers anxiety-related behaviors and to identify new treatments for anxiety-related disorders. Although substantial progress has been made and many of the key neural circuits and molecular pathways mediating stress responsiveness have been characterized, these advances have thus far failed to translate into fundamentally new treatments that are safer and more efficacious in humans. The purpose of this article is to describe methods that have been historically used for this type of research and to highlight new approaches that align with recent conceptualizations of disease symptomatology and that may ultimately prove to be more fruitful in facilitating the development of improved therapeutics.
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![Figure 1 Assessment of the acoustic startle reflex in rodents. (A) Sprague-Dawley rat in an apparatus that quantifies startle responses. Rats are placed within a nonconfining holder to minimize restraint stress. Startle response is induced by a burst of white noise delivered via a speaker located behind the rat. The force displaced is quantified by an accelerometer beneath the holder. (B) Acute effects of the stress-peptide corticotropin-releasing factor (CRF; administered intracerebroventricularly [ICV]) or vehicle only (VEH) on acoustic startle in 150-minute test sessions. Data are expressed as (%) change from pretreatment tests. CRF produces a dosedependent increase in startle. *P<0.05;**P<0.01 ; Tukey's t-tests.](/paper-figures/1cf377c1a83e4ebeb6145fc99fd6e50c/behavioral-methods-to-study-anxiety-in-rodents-figure-1.jpg)
![Figure 2 Electroencephalogram/electromyogram (EEG/EMG) telemetry in rodents as a potential index of stress. The use of telemetry in mice and rats allows derivation of numerous metrics, including sleep architecture and circadian rhythm. (A) C57BL/6 mouse next to a transmitter (manufacturer: Data Sciences International) that is subcutaneously implanted to allow remote measurements: EEG/EMG, activity, and body temperature. (B) Representative example of a brief period of EEG and EMG signal from awake, slow-wave sleep, or rapid eye movement (REM) sleep. (C) Regular circadian fluctuations of activity and temperature in C57BL/6J mice, where 0-12 h is the light phase and 12-24 h is the dark phase. These cycles can be dysregulated by stress. (D) Effects of the stress peptide pituitary adenylate cyclase-activating polypeptide (PACAP; administered intracerebroventricularly [ICV]) on exploratory behavior in the elevated plus maze (EPM) (left) and REM sleep (right) in C57BL/6J mice. PACAP decreased exploratory behavior and increased REM during the dark phase, au, arbitrary units; ZT, zeitgeber time](/paper-figures/21613f59d70c4c3ba713cd09759fad89/behavioral-methods-to-study-anxiety-in-rodents-figure-2.jpg)
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“Symptoms of mental illnesses are accompanied by emotional, cognitive, and motivational aspects that are not attributable to the lower mammal species being studied (Lezak et al., 2017).”
From The Epigenetics of Anxiety Pathophysiology: A DNA Methylation and Histone Modification Focused Review (2023) · cites this paper as Lezak et al., 2017
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“Acoustic startle is a measurable “flinch” to a delivered white noise, that has an amplitude that can be quantified in units of force, and is hypothesized to reflect the state of alertness associated with increased anxiety (Lezak et al., 2017).”
From The Epigenetics of Anxiety Pathophysiology: A DNA Methylation and Histone Modification Focused Review (2023) · cites this paper as Lezak et al., 2017
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“For example, pairing an aversive unconditioned stimuli, such as a footshock, with a tone/sound, light, or a context/environment (neutral conditioned stimuli) can elicit fear responses to the conditioned stimuli in the absence of the footshock on re-exposure to the conditioned stimulus (Lezak et al., 2017).”
From The Epigenetics of Anxiety Pathophysiology: A DNA Methylation and Histone Modification Focused Review (2023) · cites this paper as Lezak et al., 2017
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Zhu, S., Saeki, K., Yamada, L., Karlsson, R., Abebe, D., & Ozato, K. (2026) · Brain Behavior & Immunity - Health · doi:10.1016/j.bbih.2026.101395
Metadata
- Authors
- Lezak, K. R., Missig, G., & Carlezon Jr, W. A.
- Affiliations
- Harvard University (US)
- Behavioral Genetics Laboratory, Department of Psychiatry, McLean Hospital, Harvard Medical School, Belmont, Massachusetts, USA
- Venue
- Dialogues in Clinical Neuroscience
- Publisher
- Informa UK Limited
- Published
- 2017-06-30
- Volume
- 19
- Issue
- 2
- Pages
- 181-191
- ISSN
- 1294-8322 (ISSN-L)
- 1958-5969
- PMC
- PMC5573562
- OpenAlex
- W2984369896
- Cited by
- 290
- References
- 100
- Citations by year
- 2026: 16 · 2025: 39 · 2024: 35 · 2023: 45 · 2022: 47 · 2021: 39 · 2020: 36 · 2019: 25 · 2018: 7 · 2017: 1
- FWCI
- 10.27
- Type
- journal article
- Language
- en
- Open access
- diamond
- doi.org
- Licence
- Locations
- Dialogues in Clinical Neuroscience (publishedVersion, cc-by-nc-nd)
- PubMed Central (publishedVersion, cc-by-nc-nd)
- PubMed (publishedVersion)
- Funders
- National Institutes of Health (5r01mh063266-25)
- National Institute of Mental Health (R01-MH063266)
- Topics
- Life Sciences › Neuroscience › Behavioral Neuroscience › Stress Responses and Cortisol
- Life Sciences › Neuroscience › Cellular and Molecular Neuroscience › Neurotransmitter Receptor Influence on Behavior
- Life Sciences › Neuroscience › Cognitive Neuroscience › Memory and Neural Mechanisms
- Keywords
- anxiety, stress, anxiety-related disorders, stress responsiveness, translational research, behavioral tests
- MeSH terms
- Animals, Anxiety Disorders, Behavior, Animal, Disease Models, Animal, Biomedical Research, Mice, Rats, Translational Research, Biomedical
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