Behavioral Neuroscience

© 2018 American Psychological Association 2018, Vol. 132, No. 6, 580 –586
0735-7044/18/$12.00 http://dx.doi.org/10.1037/bne0000262

Acetaminophen Disrupts Memory in Object Recognition and Increases

Extracellular Signal-Regulated Kinase Phosphorylation in Male Mice
Tyler M. Milewski and Patrick T. Orr
University of Scranton

Acetaminophen (also known as paracetamol) is a commonly used over-the-counter pain medication, but
recent evidence suggests that a single exposure or prenatal exposure may have significant behavioral
effects. This investigation aimed to determine whether acetaminophen could disrupt memory formation
This article is intended solely for the personal use of the individual user and is not to be disseminated broadly.

in an object-recognition task and to quantify potential changes in memory-related signaling cascades in

This document is copyrighted by the American Psychological Association or one of its allied publishers.

the hippocampus of mice after acetaminophen administration. Using male mice, we examined the effect
of a single subcutaneous injection of acetaminophen on the object-recognition task, a single-trial,
hippocampus-dependent memory task. We also investigated potential changes in the activation of
extracellular signal-regulated kinase (ERK) in the dorsal mouse hippocampus 1 hr after a subcutaneous
injection of acetaminophen. We found that 50 mg/kg and 100 mg/kg interfered with performance in the
object-recognition memory task, whereas 10 mg/kg did not. We also found that a single 50 mg/kg
injection of acetaminophen significantly increased p42 ERK phosphorylation in the dorsal mouse
hippocampus. Overall, these results suggest that a single dose of acetaminophen can have significant
effects on memory and alters signaling kinases critical for memory consolidation. Further work is needed
to determine the involved mechanisms.

Keywords: acetaminophen, object recognition, ERK, hippocampus, paracetamol

Acetaminophen, also known as paracetamol, is one of the most
commonly used over-the-counter pain medications. Recom-
mended as a safe, mild analgesic (Källén & Reis, 2016; Servey &
Chang, 2014), acetaminophen is frequently used for postopera-
tional pain in combination with a nonsteroidal anti-inflammatory
drug (Koh, Nguyen, & Jahr, 2015; McNicol, Ferguson, Haroutou-
nian, Carr, & Schumann, 2016; Ong, Seymour, Lirk, & Merry,
2010) for dealing with the pain of fractures and broken bones
(Bergenstock, Min, Simon, Sabatino, & O’Connor, 2005) and
migraines (Lipton, Baggish, Stewart, Codispoti, & Fu, 2000). In
addition, acetaminophen is the analgesic of choice during preg-
nancy because it does not confer risk for frank congenital abnor-
malities (Källén & Reis, 2016; Scialli, Ang, Breitmeyer, & Royal,
2010). However, recent evidence suggests that acetaminophen
may have an impact beyond its analgesic and antipyretic effects.
Recent prospective studies link prenatal acetaminophen expo-
sure to symptoms of hyperactivity or impulsivity (Avella-Garcia et
al., 2016), poorer gross motor development (Brandlistuen, Ystrom,
Nulman, Koren, & Nordeng, 2013), and increased risk of hyper-
This article was published Online First August 30, 2018.
Tyler M. Milewski and Patrick T. Orr, Department of Psychology,
Neuroscience Program, University of Scranton.
We thank J. Timothy Cannon for aid in the conception of the study, and
Shreya Patel, Catherine Stapf, and Sara Wierbowski for assistance in
collecting behavioral data.
Correspondence concerning this article should be addressed to Patrick T.
Orr, Department of Psychology, Neuroscience Program, University of
Scranton, Rear 815 Ridge Row, 200 Alumni Memorial Hall, Scranton, PA
18510. E-mail: patrick.orr@scranton.edu
kinetic disorders (Liew, Ritz, Rebordosa, Lee, & Olsen, 2014) or
autism spectrum disorders with hyperkinetic symptoms (Liew,
Ritz, Virk, & Olsen, 2016). In acute administration studies of
humans, acetaminophen use has also been reported to dull emo-
tional pain (DeWall et al., 2010) and inhibit emotional processing
(Durso, Luttrell, & Way, 2015). Further, a single acute dose of
acetaminophen has been found to reduce distrust in individuals
with features of borderline personality disorder (Roberts, Krajbich,
Cheavens, Campo, & Way, 2018). In light of these findings,
careful investigation is needed to determine if acetaminophen may
have additional underdescribed effects.
Recent rodent work demonstrates that acetaminophen affects
memory. For example, injections of high doses of acetaminophen
have impaired performance of the Morris water maze in mice,
whereas low doses facilitated performance (Ishida et al., 2007).
Acetaminophen has also interfered with long-term potentiation in
the hippocampus (Chen & Bazan, 2003). Although such evidence
suggests that acetaminophen can influence memory formation, the
mechanism for this action remains unclear. It is currently unknown
whether acetaminophen exposure influences object-recognition
memory formation, but, as a single-trial learning task, it is ideal for
testing the effects of acute acetaminophen exposure.
Some investigations suggest that the effects of acetaminophen
are mediated by anti-inflammatory actions, such as inhibition of
cyclooxegenase (COX) enzymes (Ishida et al., 2007), notably
COX-2 and COX-3 (although, for a full discussion of the putative
COX-3, see Kis, Snipes, & Busija, 2005). Inhibition of COX-2 is
known to impair hippocampus-dependent memory consolidation
in rats (Teather, Packard, & Bazan, 2002). This mechanism could
be mediated in some way by serotonin (Chen & Bazan, 2003;
Ishida et al., 2007). Alternatively, the mnemonic effects of acet-

580
 ACETAMINOPHEN DISRUPTS OBJECT-RECOGNITION MEMORY
aminophen could be mediated by actions on the cannabinoid
system. N-Arachidonoylphenolamine (AM404), a metabolite of
acetaminophen (Högestätt et al., 2005), acts as a cannabinoid
agonist via inhibition of cannabinoid reuptake (Fegley et al., 2004;
Glaser et al., 2003), and these pathways have been linked with
memory consolidation. Direct injection of AM404 inhibits contex-
tual fear memory, whereas injections of the cannabinoid receptor
antagonist, AM481, promotes contextual fear memory (Lin et al.,
2011). Further, agonists of cannabinoid receptor CB1 interfere
with object-recognition memory consolidation (Clarke et al.,
2008). CB1 receptors are found densely within the hippocampus
(Kawamura et al., 2006), which is critically important for both
contextual fear memory (Ruehle, Rey, Remmers, & Lutz, 2012)
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and object recognition (Baker & Kim, 2002; Clark, Zola, & Squire,
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2000; Cohen et al., 2013; Cohen & Stackman, 2015; Stackman,
Cohen, Lora, & Rios, 2016). In addition, there are significant
connections between COX-2 and endocannabinoid signaling (for
review, see Yang & Chen, 2008), suggesting that these two po-
tential mechanisms downstream of acetaminophen may not be
cleanly separable.
The extracellular signal-regulated kinase/mitogen activated pro-
tein kinase (ERK/MAPK) signaling cascade is a well-established
pathway necessary for multiple hippocampus-dependent tasks, in-
cluding object recognition (Fernandez et al., 2008), the Morris
water maze (Selcher, Atkins, Trzaskos, Paylor, & Sweatt, 1999),
and contextual fear conditioning (Schafe et al., 2000). The ERK/
MAPK pathway, which initiates cyclic adenosine monophosphate-
response element-binding protein (CREB)-related gene transcrip-
tion (Sweatt, 2001), is necessary for both late-phase long-term
potentiation (Ying et al., 2002) and metabotropic glutamate recep-
tor long-term depression (Sanderson, Hogg, Collingridge, & Cor-
rêa, 2016) in the hippocampus. Of interest, although cannabinoid
signaling interferes with hippocampus-dependent memory tasks, it
increases activation of the ERK/MAPK pathway both in vitro
(Derkinderen et al., 2003) and in vivo (Basavarajappa, Nagre, Xie,
& Subbanna, 2014; Rubino, Forlani, Viganò, Zippel, & Parolaro,
2004). Actions of COX-2, on the other hand, tend to be associated
with p38 MAPK, rather than p42/p44 (ERK1 and ERK2; Yang &
Chen, 2008). Examining the effect of acetaminophen administra-
tion on hippocampal ERK activation may be useful in teasing apart
these potential mechanisms of action. In this study, we examined
the effects of multiple doses of acetaminophen injections on mem-
ory in the object-recognition task in mice and subsequently exam-
ined ERK phosphorylation in the dorsal hippocampi of mice
receiving acetaminophen injections. We found that acetaminophen
interferes with memory consolidation and increases ERK phos-
phorylation in the dorsal hippocampus.
Method and Materials
Subjects
Male C57BL/6 mice (N ⫽ 57) were obtained from the Univer-
sity of Scranton Neuroscience Mouse Breeding Colony at 8 to 12
weeks of age. All mice were housed in individual cages with ad
libitum access to food and water on a 12/12 light– dark cycle
(lights on at 7:00 a.m.). All behavioral testing was conducted
during the light phase of the cycle. All procedures were approved
581
by the institutional Animal Care Use Committee of the University
of Scranton.
Object Recognition
Object recognition was performed to assess nonspatial hip-
pocampal memory (Baker & Kim, 2002; Clark et al., 2000; Fer-
nandez et al., 2008). This is a one trial learning task which is,
consequently, ideal for examining the effects of acute drug admin-
istration on hippocampal memory consolidation. This task, which
relies on novelty preferences inherent in mice, consists of three
phases, habituation, training, and testing, all of which take place in
an evenly lit, white wooden box (60 cm wide ⫻ 60 cm long ⫻ 68
cm high). Each phase was separated by 24 hr. During the habitu-
ation phase, each mouse was placed into the empty chamber and
allowed to freely explore for 5 min. This was primarily to allow for
the mouse to become accustomed to the box, but grid crossings
during this phase were scored to ensure there were no group
differences in locomotor behavior prior to the experimental ma-
nipulation. For this analysis, a 4 ⫻ 4 grid was superimposed over
the testing chamber and grid crossings were recorded. In the
second phase, training, each mouse was placed into the chamber
with two identical copies of an object positioned near the corners
of one side. The mouse was started facing the wall opposite the
objects and was allowed to freely explore the environment and
objects until it accrued a total of 30 s of object-exploration time
(summed across both copies of the object). Holding constant the
total object-exploration time rather than the total time spent in the
box during this phase ensures that the mice have similar memory-
trace strengths. Immediately after phase two, mice received a
subcutaneous injection of the vehicle or one of three doses of
acetaminophen, as described below. During the third phase, test-
ing, the mouse was placed into the box along with a novel object
and one copy of the object used during training. The mouse was
allowed to freely explore the box and the objects for 10 min, and
time spent with each object was recorded. Mice have a natural
affinity for novelty. If a mouse spent more time with the novel
object, it implies that the mouse had a memory of the familiar
object.
During this task the two objects used were a yellow plastic box
and a faucet, each approximately 6 –7 cm wide and high. The
objects and box were cleaned with 70% ethanol between all mice
and between all phases. The novel object (block or faucet) and side
that the novel object was presented on were counterbalanced
throughout the experiment. All mice were returned to their home
cages between phases. Visual records of all phases were made
using a ceiling-mounted digital camera, which were stored on a
computer for later analysis. Object exploration was defined as
object-directed behavior involving contact with the snout or fore-
paws. Any mouse that took longer than 25 min to accrue 30 s of
object exploration during training was eliminated from the study.
This task was scored using a custom written stopwatch program
that halted as soon as criterion was met. Judgments of exploration
were made by a human observer trained in the behavior.
Drug Administration
Following the training phase of object recognition, each mouse
was randomly assigned to receive a subcutaneous injection of the
 582 MILEWSKI AND ORR
vehicle or one of three doses of acetaminophen (MilliporeSigma
#103–90-2, Burlington MA). The vehicle (n ⫽ 14) consisted of
0.1% 2-hydroxy-␤-cyclodextrin (HBC, MilliporeSigma #128446 –
35-5) in saline. Acetaminophen doses were 10 mg/kg (n ⫽ 14), 50
mg/kg (n ⫽ 15), or 100 mg/kg (n ⫽ 14) acetaminophen in 0.1%
HBC-saline. These doses are based on doses originally used in rats
(Blecharz-Klin et al., 2013) to affect cognition and were not
intended for therapeutic analgesic effect. Previous work has indi-
cated that hepatotoxicity due to acetaminophen begins at 150
mg/kg (Liu et al., 1999), so these doses should not have been
hepatotoxic for these mice.
Western Blotting
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After behavioral testing, a separate group of 21 mice was
injected with the vehicle (n ⫽ 11) or the lowest behaviorally
effective dose of acetaminophen (50 mg/kg; n ⫽ 10) and sacrificed
one hour after injection. Each dorsal hippocampus was immedi-
ately dissected out of the brain and stored at ⫺80 °C until homog-
enization. Briefly, each dorsal hippocampal sample was suspended
in 1:50 weight/volume dilution of a lysis buffer and homogenized
with a probe sonicator (Misonix, Newtown, CT), then suspended
in the sample buffer and boiled for 5 min. The homogenate was
aliquoted to a concentration of 2 ␮g/␮l. Aliquots and homogenates
were stored at ⫺80 °C until Western blotting. Aliquots were then
electrophoresed on 10% tris-hydrochloride gels and transferred to
an Immobilon polyvinylidene difluoride (PVDF) membrane.
Western blots were blocked in 10% milk and incubated overnight
with antiphospho-p44/p42 ERK primary antibodies (1:2000; Cell
Signaling, Danvers, MA). Blots were then incubated with anti-
rabbit-horseradish peroxidase (HRP) (1:20,000; Cell Signaling)
and developed using West Dura chemiluminescent (CL) substrate
(SuperSignal West Dura, ThermoFisher, Rockford, IL). Blots were
imaged using CL-Xposure film (ThermoFisher #34090) and den-
sitometry was conducted using ImageJ (https://imagej.nih.gov/).
Following imaging, blots were stripped using 0.2M sodium hy-
droxide and reprobed with antitotal-p44/p42 ERK (1:100; Cell
Signaling). Phosphorylated ERK (pERK) levels were normalized
to total ERK (tERK) levels, respectively, and expressed as percent
immunoreactivity relative to the vehicle controls.
Data Analysis
Data analyses were conducted using IBM SPSS 23 (IBM Cor-
poration, Armonk, NY). In object recognition, an exploration ratio
was calculated for the training and testing phases. For the testing
phase, this ratio was the time spent exploring the novel object
divided by the total object-exploration time (time spent exploring
the novel object plus time spent exploring the familiar object). In
the training phase, this ratio was the time spent exploring the
object presented on the left side of the chamber divided by the total
object-exploration time (time spent exploring both copies of the
object). If object exploration is due to chance, then these ratios
would be expected to be approximately .5. Thus, for each group,
one-sample t tests were performed to determine if the exploration
ratio was significantly different from chance (.5). One-way anal-
ysis of variance (ANOVA) was also used to check for between-
group differences in exploration ratios. To check for preexisting
group differences, grid crossings during habituation and explora-
tion ratio during training were compared across groups using
one-way ANOVAs. For Western blotting data, an independent-
samples t test was used to check for differences in the normalized
pERK/tERK values for the control and 50 mg/kg group.
Results
Object Recognition
Mice did not show a significant side preference during training
(Figure 1A). The training exploration ratio did not differ signifi-
cantly from .5 for the control, t(13) ⫽ ⫺.152, p ⫽ .881, 10 mg/kg,
t(13) ⫽ 1.831, p ⫽ .09, 50 mg/kg, t(14) ⫽ .124, p ⫽ .903, or 100
mg/kg, t(13) ⫽ ⫺0.347, p ⫽ .734. In addition, the training:
exploration ratio did not differ significantly across groups, F(3,
53) ⫽ 0.89, p ⬎ .05. There were not significant differences
between groups for locomotor activity during habituation (see
Table 1). Groups did not significantly differ on total grid crossings,
F(3, 53) ⫽ 0.499, p ⬎ .05 or on crossings in the inner, F(3, 53) ⫽
1.664, p ⬎ .05 or outer, F(3, 53) ⫽ 0.466, p ⬎ .05 portions of the
chamber. Overall, these results suggest that there were no differ-
ences in the mice prior to injection, and that the differences
observed during testing were due to the effects of acetaminophen.
Acetaminophen affected object-recognition memory consolida-
tion. Mice receiving subcutaneous injection of the vehicle and 10
mg/kg of acetaminophen showed a significant preference for the
familiar object, whereas the mice receiving 50 mg/kg and 100
mg/kg injections did not (Figure 1B). The novel exploration ratio
was significantly higher than .5 for the vehicle, t(13) ⫽ 2.419, p ⫽
.031 and 10-mg/kg, t(13) ⫽ 2.98, p ⫽ .011 groups. There was no
significant difference from .5 for the 50-mg/kg, t(14) ⫽ ⫺1.455,
p ⫽ .168 or 100-mg/kg group, t(13) ⫽ ⫺.102, p ⬎ .05. Further, a
one-way ANOVA comparing the exploration ratios of these
groups was significant, F(3, 53) ⫽ 3.155, p ⫽ .032. Tukey’s
honestly significant difference post hoc testing revealed that the
50-mg/kg group was significantly different from the control group
(p ⫽ .05). This pattern of data suggests that a single posttraining
subcutaneous injection of acetaminophen at 50 and 100 mg/kg, but
not 10 mg/kg, is sufficient to disrupt object-recognition memory.
Western Blotting
Subcutaneous administration of acetaminophen affected p42
ERK phosphorylation (see Figure 2). One hr after subcutaneous
injection, dorsal hippocampal levels of phosphorylated p42 ERK
were significantly higher in the group receiving 50 mg/kg acet-
aminophen than in the vehicle-treated group, t(19) ⫽ 2.219, p ⫽
.039. There was not a significant difference between groups in
phosphorylated p44 ERK levels, t(19) ⫽ 0.866, p ⬎ .05.
Discussion
In this investigation, a single subcutaneous posttraining injec-
tion of 50 mg/kg or 100 mg/kg acetaminophen interfered with
memory consolidation in the object-recognition task and a subcu-
taneous injection of 50 mg/kg led to a significant increase in dorsal
hippocampal p42 ERK phosphorylation. These data are the first to
demonstrate an impairment of hippocampal-dependent memory
consolidation by acetaminophen. Although acetaminophen has
 ACETAMINOPHEN DISRUPTS OBJECT-RECOGNITION MEMORY 583
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Figure 1. Exploration preferences during training and testing, by drug condition. For all data shown, each bar
represents the mean ⫾ SEM, and the dashed line represents .5, which is chance performance. (A) For training,
the exploration ratio represents time spent exploring the object on the left, divided by the total time spent
exploring objects during training. During training, no groups showed a significant side preference. (B) For
testing, the exploration ratio represents the time spent exploring the novel object, divided by the total amount of
time spent exploring objects during testing. Asterisks indicate that a group is significantly different from chance
performance (p ⱕ .05) and dagger indicates a significant difference between indicated groups (p ⱕ .05). Groups
receiving the vehicle or 10 mg/kg acetaminophen spent significantly more time with the novel object during
testing, whereas the groups receiving 50 mg/kg or 100 mg/kg did not, demonstrating an impairment in memory.

been reported to interfere with Morris water-maze performance
(Ishida et al., 2007), the multiple learning and testing trials, along
with the multiple injections of acetaminophen in that study, make
the effect of acetaminophen unclear. That is, because acetamino-
phen was present throughout the entire task, the effect observed
could be an effect on acquisition or some other nonmnemonic
aspect of task performance. Thus, it is not clear that this previously
reported effect was an effect on memory consolidation. However,
in the current investigation, acetaminophen exposure was limited
to a single injection immediately following training, suggesting
that the effects of acetaminophen were likely limited to consoli-
dation. The half-life of acetaminophen in humans is on the order of
2–2.5 hr (Forrest, Clements, & Prescott, 1982), which is consistent
with interpreting this as a limited, consolidation-only exposure.
These data are also the first evidence that acetaminophen can
drive p42 ERK phosphorylation. This connection between acet-
aminophen and a signaling molecule critical for learning and
memory (Sweatt, 2001) suggests that acetaminophen could have a
significant impact on multiple types of learning tasks, a possibility
that needs further investigation. The increase in ERK phosphory-
lation downstream of acetaminophen is also interesting because it
is concordant with the effects on ERK of cannabinoids (Basavara-
jappa et al., 2014; Derkinderen et al., 2003; Rubino et al., 2004),
a potential downstream effector of acetaminophen.
This correspondence is similar to the pattern of results observed
in other behavioral paradigms. AM404 injections reduce the dis-
play of anxiety in rats in a manner dependent on CB1 receptors
(Bitencourt, Pamplona, & Takahashi, 2008; Bortolato et al., 2006).
Acute intraperitoneal injections of acetaminophen and AM404
also reduce the display of anxiety in mice in a CB1-dependent
manner (Umathe, Manna, Utturwar, & Jain, 2009). Further, the
anxiolytic actions of acetaminophen injection in mice are depen-
dent on the activity of the fatty acid, amide hydrolase, which is
responsible for the synthesis of AM404 (Zaitone, El-Wakeil, &
Abou-El-Ela, 2012). It is possible that the effects of acetamino-
phen on hippocampal memory consolidation depends on the same

Table 1
Additional Behavioral Data Collected During Object Recognition

Group Total grid crossings Inner grid crossings Outer grid crossings

Vehicle 158.79 ⫾ 11.433 25.93 ⫾ 3.146 132.86 ⫾ 12.258

10 mg/kg 168.86 ⫾ 8.915 21.21 ⫾ 2.136 147.64 ⫾ 7.778
50 mg/kg 167.60 ⫾ 9.041 23.87 ⫾ 1.732 143.73 ⫾ 8.958
100 mg/kg 176.57 ⫾ 11.352 30.50 ⫾ 4.463 146.07 ⫾ 9.316
Note. Values represent the mean ⫾ SEM.
 584 MILEWSKI AND ORR
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Figure 2. Phosphorylated/total ERK in the dorsal hippocampus of mice receiving the control vehicle or 50
mg/kg acetaminophen. All values were normalized to the mean for the vehicle-treated group. Each bar represents
the group mean ⫾ SEM. Asterisks indicate significant group differences (p ⱕ .05). Mice receiving 50 mg/kg
acetaminophen had significantly higher levels of phosphorylated p42 ERK relative to total p42 ERK. There were
no significant differences in p44 ERK. Representative phosphorylated ERK bands are inset. The inset image was
scaled up for this figure for clarity with the open-source GIMP software (GNU Project, Boston, MA) using cubic
interpolation, but analyses were conducted on the original, unaltered images.

mechanisms, although the current data do not offer any direct
evidence. Further investigation is needed to determine whether the
effects if acetaminophen on memory depend on CB1 signaling.
In this study, the observed changes in ERK phosphorylation
occurred in the dorsal hippocampus, which is not surprising, given
that this form of the object-recognition task depends on ERK
signaling in the dorsal hippocampus (Fernandez et al., 2008).
Although the changes in ERK phosphorylation are compelling, the
current data did not demonstrate that this ERK phosphorylation is
necessary for the acetaminophen-induced impairment in this mem-
ory task. Future experiments are required to determine if this
change in ERK phosphorylation in the dorsal hippocampus is a
downstream mechanism critical to the effect of acetaminophen on
hippocampal memory consolidation or if it is an unrelated phe-
nomenon.
In the current investigation, acetaminophen increased p42 ERK
phosphorylation but not p44 ERK phosophorylation. Previous
work suggests that p42 and p44 ERK may play different signaling
roles (Lloyd, 2006). These isoforms regulate distinct sets of genes
in zebrafish (Krens, Corredor-Adamez, He, Snaar-Jagalska, &
Spaink, 2008). In rodents, modulation of p42, but not p44, ERK is
associated with enhancement of memory by steroid hormones
(Fernandez et al., 2008; Orr, Rubin, Fan, Kent, & Frick, 2012).
Further, Selcher, Nekrasova, Paylor, Landreth, and Sweatt (2001)
observed no differences in mice lacking p44 ERK. Thus, the
current findings further reinforce that p42 ERK appears to be the
ERK isoform most relevant to memory formation.
Notably, previous work indicates that hepatoxicity due to acet-
aminophen begins at 150 mg/kg (Liu et al., 1999), so the doses of
acetaminophen that inhibited memory formation in the current
study are not hepatotoxic. The effective doses are somewhat
smaller than the dose (302.3 mg/kg) that has been shown to inhibit
water-maze performance (Ishida et al., 2007), and are comparable
to the maximum daily dose for an adult human (50 – 60 mg/kg),
suggesting that these findings are ethologically valid.
Overall, the results indicate that a single dose of acetaminophen
can impair memory consolidation and drive ERK phosphorylation
in the dorsal hippocampus. Further work is needed to determine
the downstream mechanisms of this effect and the potential effects
on humans.
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Received February 25, 2018
Revision received May 25, 2018
Accepted May 31, 2018 䡲