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Received: 1 April 2021 
|
  Revised: 13 April 2021 
|  Accepted: 29 April 2021

DOI: 10.1111/ejn.15270

CO M M E N TA RY

The forced swim test: Giving up on behavioral despair

(Commentary on Molendijk & de Kloet, 2021)

Erin Gorman-­Sandler   | Fiona Hollis

Department of Pharmacology, Physiology, and Neuroscience, University of South Carolina School of Medicine, Columbia, SC, USA

Correspondence
Fiona Hollis, Department of Pharmacology, Physiology, and Neuroscience, University of South Carolina School of Medicine, Columbia, SC, USA.
Email: fiona.hollis@uscmed.sc.edu

The article that is the subject of this Commentary is available here: 10.1111/ejn.15139

Funding information
National Institute of General Medical Sciences, Grant/Award Number: P20GM103641

Keywords: animal model, behavior, brain, depression, forced swim test, FST, stress

CO M M EN TA RY
Depression is a prevalent, debilitating mental illness which
affects over 300 million people worldwide and is often co-
morbid with chronic illnesses or anxiety disorders (Ménard
et  al.,  2016; World Health Organization (WHO), 2017).
Characterized by a wide range of symptoms involving dis-
ruption of mood and cognition, depression is a leading cause
of disability and contributor to economic burden (Ménard
et  al.,  2016; World Health Organization (WHO), 2017).
Even as incidence of depression steadily escalates (Akil
et  al.,  2018), research efforts to understand its increasingly
complex pathology and produce more effective treatments
are stymied. The symptoms of depression are heterogeneous
and do not point to a single etiological origin (Nestler &
Hyman, 2010). While animal models of depression and their
accompanying behavioral validations are fundamental to de-
pression research, they can also be a source of frustration. The
Forced Swim Test (FST, also known as the Porsolt swim test)
in particular has proven controversial. Developed by Roger
Porsolt in the 1970s as a rapid behavioral screen for antide-
pressant compounds (Porsolt et al., 1977, 1978), the FST is
now considered by some as “the gold standard animal test for
depression” (Unal & Canbeyli, 2019). The classic FST is a 2-­
day test where a rodent is placed in an inescapable container
Abbreviations: FST, forced swim test.
of water and must swim for 15  min on the first day and 5
or 6  min on the second “test” day twenty-­four hours later
(Porsolt et al., 1977, 1978; Yankelevitch-­Yahav et al., 2015;
Figure 1). During the test session, animals quickly begin to
float, exhibiting immobility behavior that has been inter-
preted as behavioral despair, learned helplessness, passive
coping, psychomotor retardation, anxiety, and even autism.
But what is the FST really measuring? Does immobility in
this test really correspond to behavioral despair, serving as a
reliable measure of depressive-­like behavior?
These are the questions that Molendijk and de Kloet thor-
oughly explore in their latest review in the Special Issue of
European Journal of Neuroscience. Having followed the
FST usage and interpretations for at least the past 6  years,
Molendijk and de Kloet are key contributors in promoting
discussions surrounding the interpretation of the FST. The
current review emphasizes their previous work and continues
to track FST interpretations in the field. Additionally, they
further expand on usage of the FST and potential mecha-
nisms behind its observed behaviors. Overall, the article pro-
vides unique and detailed perspectives using meta-­analyses
of recent FST literature.
The authors begin by providing an update on the trends in
the usage and interpretation of the FST since their previous
assessments in 2015 (Molendijk & de Kloet, 2015) and 2019
(Molendijk & de Kloet, 2019). Molendijk and de Kloet com-
piled and analyzed datasets of recent publications between

© 2021 Federation of European Neuroscience Societies and John Wiley & Sons Ltd

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COMMENTARY 2833

F I G U R E 1   Methodology for classic forced swim test in rodents. (a) Typically, rats will first undergo a 15-­min swim exposure (water
temperature ≈24 ± 2°C) which is termed the “pre-­test.” After 24 h, the 5-­min forced swim “test” is conducted. Alternatively, rats may undergo a
single 15-­min swim exposure depending on the goals of the experiment. For example, if the goal is to measure retention of acquired immobility,
the classic 2-­day test should be used. (b) Mice usually undergo a single 6-­min swim exposure with the first 2-­min serving as the “pre-­test” and
the last 4 functioning as the “test.” (c) Behavioral scoring may include percent time spent immobile/total time in test, time spent swimming (with
swimming often including climbing and diving actions), and/or latency (seconds) to become immobile. Immobility is also defined as “floating” or
using minimal actions to keep the head above water

2018 and 2020 which use the FST, describing updated statis-
tical assessments of trends in usage and interpretations over
time, across countries, in different journals, and between dif-
ferent behavioral outcomes. Here, they found that the use of
the FST in publications is no longer increasing, indicating
a shift in trend for the first time since the FST's inception.
Pooling from a random selection of articles, the authors re-
port a decline in interpreting FST immobility as depressive-­
like, particularly from European and North American
regions. Scoring immobility as coping or “other” exhibited a
slight increase, with a similar relationship to geographical re-
gion (Molendijk & de Kloet, 2019). Molendijk and de Kloet's
analysis takes the pulse of the global scientific community's
usage of the FST, allowing us to make predictions about the
test's future in depression research.
Molendijk and de Kloet then assembled a second data-
set of FST literature from five behaviorally focused journals
with large amounts of relevant publications to determine
whether studies measuring different behaviors observe over-
lap in their outcomes. Frequently, individual studies will
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2834
use a series of different behavioral tests in combination as
to provide a broader picture of an animal's or group's be-
havioral phenotype (Molendijk & Kloet,  2021; Nestler &
Hyman, 2010). Thus, this review analyzed overlap between
use of the Tail Suspension Test, the Sucrose Preference Test,
and anxiety-­like measures (Elevated Plus Maze, Light-­Dark
Box, Open Field Test), in conjunction with the FST. These
data provide fascinating insight into individual variability in
behavior within groups. That is, an anhedonic phenotype in
the Sucrose Preference Test will not always positively cor-
relate with passive coping in the FST or anxiety-­like behav-
iors in the Elevated Plus Maze, and vice versa. While it is
convenient to assume that, for example, a chronically stressed
animal will present with depressive-­and anxiety-­like be-
haviors across each test, this is certainly not the outcome in
many cases. Importantly, this analysis can also inform our
understanding of different interpretations of immobility. For
instance, the lack of correlation between anxiety-­like behav-
iors in several different tests and FST immobility suggests
that the FST is not measuring anxious modalities.
Another original aspect of the current review lies in the
section where the authors gathered opinions from the editors
of previously mentioned journals which frequently publish
FST-­related articles. Here, they asked for a point of view
from each editor on submissions which describe FST immo-
bility as depressive-­like behavior. While one journal provided
no opinion, most editors tend to give their trusted reviewers
the larger say in whether submissions are reaching appropri-
ate conclusions. However, most of the editors also provided
opinions reflecting the idea that the FST and other behavioral
outcomes should be interpreted with caution and not be relied
on too greatly. Editors-­in-­Chief Dr. Heuser and Dr. Dantzer
of Psychoneuroendocrinology specifically emphasize that
authors should examine behavioral guidelines put forth by the
National Institute of Mental Health (NIMH). Further on in
the review, Molendijk and de Kloet address these guidelines
and discuss the NIMH Research Domain Criteria (RDoC).
These criteria call researchers to consider multiple levels of
disease complexity (such as genetics, behavior, or circuitry)
when studying aspects of psychiatric disorders, with an over-
arching goal of discovering mechanisms which may translate
to human conditions. Importantly, we should recognize that
we are individually studying aspects of a more dimensional
problem, rather than “models” which tend to reduce multi-
faceted psychiatric disorders to a single behavioral or molec-
ular difference. As higher governing institutions and many
well-­respected journals are moving towards these ideas, re-
viewers and authors might also become more deliberate in
their experimental analyses and interpretations. With these
considerations in mind, we can continue our research with
the knowledge that we are contributing to a greater mission
of integrating relevant information in the psychiatric field.
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COMMENTARY
The authors finally review the literature, providing a thor-
ough background of interpretations of the FST over time,
including its limited use as a measure for validation of antide-
pressant compounds. They discuss and contrast the role of the
HPA-­axis, including both glucocorticoid and mineralocorti-
coid receptors (GRs and MRs), in stress coping behaviors.
Finally, they elegantly highlight and synthesize recent liter-
ature which suggests circuitry involved in coping strategy,
insinuating that interactions between these “bottom-­up en-
docrine” and “top-­down circuitry” processes are involved in
immobility behavioral output (i.e., coping strategy), linking
these interactions to possible genetic phenotypic differences
in different strains. Overall, these findings provide strong ev-
idence for alternative explanations underlying animal perfor-
mance in the FST.
While controversies involving the FST are prevalent
both within science and in the public, contemplating both
value and ethicality, it is important to consider the follow-
ing questions: Are current behavioral paradigms providing
us with the validity we actually need? Is the FST being
used in the correct manner to provide the most valuable
information? Furthermore, are we considering these same
issues for other behavioral paradigms? Are over-­reliant
interpretations of these behaviors restricting progress in
the field of mental health? Indications in this review and
elsewhere suggest that the FST is no longer considered the
miracle test to predictively validate antidepressant effects,
and rightly so. Perhaps, it is worthwhile to consider other
approaches to evaluate animal phenotypes and validate
antidepressant treatments in ways that more accurately
represent the heterogeneity of clinical depression. More
importantly, it is crucial for researchers to provide argu-
ments of validity in publications, describing both strengths
and weaknesses of their models so that they can be prop-
erly evaluated. In a larger sense, this review reminds us of
the imperative to continue such evaluation—­that we must
critically assess how we are interpreting complex animal
behaviors to ensure we are not overly interpreting behav-
iors or potentially missing out on alternative and fruitful
directions.
ACKNOWLEDGEMENTS
This work was supported by an NIH grant P20GM103641
as well as intramural funding from the University of South
Carolina.
CONFLICTS OF INTEREST
The authors have no conflicts to declare.
PEER REVIEW
The peer review history for this article is available at https://
publo​ns.com/publo​n/10.1111/ejn.15270.

COMMENTARY
ORCID
Erin Gorman-­Sandler  https://orcid.org/0000-0002-1374-9067
Fiona Hollis  https://orcid.org/0000-0001-6559-5736
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