Hormones and Behavior 94 (2017) 33–39

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Hormones and Behavior

journal homepage: www.elsevier.com/locate/yhbeh

Administration of an oxytocin receptor antagonist attenuates sexual

motivation in male rats
D.S. Blitzer a, T.E. Wells a, W.R. Hawley a,b,⁎
a
Franklin and Marshall College, Department of Psychology, United States
b
Edinboro University of Pennsylvania, Department of Psychology, United States

a r t i c l e i n f o a b s t r a c t

Article history: In male rats, oxytocin impacts both sexual arousal and certain types of consummatory sexual behaviors. However,
Received 12 October 2016 the role of oxytocin in the motivational aspects of sexual behavior has received limited attention. Given the role
Revised 20 March 2017 that oxytocin signaling plays in consummatory sexual behaviors, it was hypothesized that pharmacological atten-
Accepted 3 June 2017
uation of oxytocin signaling would reduce sexual motivation in male rats. Sexually experienced Long-Evans male
Available online xxxx
rats were administered either an oxytocin receptor antagonist (L368,899 hydrochloride; 1 mg/kg) or vehicle con-
Keywords:
trol into the intraperitoneal cavity 40 min prior to placement into the center chamber of a three-chambered arena
Sexual behavior designed to assess sexual motivation. During the 20-minute test, a sexually experienced stimulus male rat and a
Sexual motivation sexually receptive stimulus female rat were separately confined to smaller chambers that were attached to the
Oxytocin larger end chambers of the arena. However, physical contact between test and stimulus rats was prevented by
Oxytocin receptors perforated dividers. Immediately following the sexual motivation test, test male rats were placed with a sexually
receptive female to examine consummatory sexual behaviors. Although both drug and vehicle treated rats exhib-
ited a preference for the female, treatment with an oxytocin receptor antagonist decreased the amount of time
spent with the female. There were no differences between drug and vehicle treated rats in either general activity,
exploratory behaviors, the amount of time spent near the stimulus male rat, or consummatory sexual behaviors.
Extending previous findings, these results indicate that oxytocin receptors are involved in sexual motivation in
male rats.
© 2016 Elsevier Inc. All rights reserved.

1. Introduction of an OT receptor antagonist (Argiolas et al., 1987; Melis et al., 1999,

Oxytocin (OT) is well known for regulating a variety of social behav-
iors. For example, in male rats, oxytonergic signaling has been shown to
impact affiliative behaviors and aggression (Calcagnoli et al., 2015a,
2015b), social memory (Lukas et al., 2013), the rewarding aspects of so-
cial interactions (Ramos et al., 2015), and social defeat-induced avoid-
ance of a conspecific (Lukas et al., 2011). Interestingly, OT also
influences different types of sexual behavior in male rats (Argiolas,
1999). Male rats exhibited higher levels of sexual arousal, as indicated
by the occurrence of penile erections, following direct administration
of OT into either the ventral tegmental area (Succu et al., 2008), the
paraventricular nucleus (PVN) of the hypothalamus (Melis et al.,
1986), the posteromedial cortical nucleus of the amygdala (Melis et
al., 2009), area CA1 of the hippocampus (Melis et al., 1986), or the
subiculum (Succu et al., 2011). In accordance with these findings, penile
erections were reduced following either intraventricular administration
⁎ Corresponding author at: Edinboro University of Pennsylvania, Department of
Psychology, Compton Hall, Room 106, 210 East Normal St., Edinboro, PA 16444, United
States.
E-mail address: whawley@tulane.edu (W.R. Hawley).
1992) or lesions of the PVN (Liu et al., 1997), which is partially com-
prised of oxytonergic neurons (George et al., 1976).
In addition to its role in sexual arousal, OT is involved in the consum-
matory aspects of male rat sexual behavior. Levels of OT in blood plasma
increased following sexual experience in both sexually naive male rats
and male rats with previous sexual experience (Hillegaart et al.,
1998). Furthermore, levels of OT in the cerebrospinal fluid have been
shown to increase two to three-fold after ejaculation (Hughes et al.,
1987; Waldherr and Neumann, 2007), an effect abrogated by lesions
of the PVN (Hughes et al., 1987). Sexual experience increased OT recep-
tor mRNA and protein expression in the medial preoptic area (MPOA),
the area of the brain that is critical for the execution of consummatory
sexual behaviors (Gil et al., 2013). As might be expected, OT adminis-
tered either centrally or peripherally decreased the number of mounts
(Gil et al., 2011) and intromissions (Stoneham et al., 1985) prior to ejac-
ulation, reduced the latency to first mount and first intromission (Arletti
et al., 1990), as well as the ejaculation latency and the post ejaculatory
interval (Arletti et al., 1992, 1990, 1985; Gil et al., 2013, 2011). Con-
versely, central administration of an OT receptor antagonist, either
into the medial preoptic area or ventricles, resulted in a longer latency
to first intromission (Gil et al., 2011), a reduction in both mounts and

http://dx.doi.org/10.1016/j.yhbeh.2017.06.002
0018-506X/© 2016 Elsevier Inc. All rights reserved.
34 D.S. Blitzer et al. / Hormones and Behavior 94 (2017) 33–39
intromissions, as well as a reduction in the likelihood of ejaculation
(Argiolas et al., 1988). Collectively, these results indicate that OT influ-
ences the consummatory aspects of male rat sexual behavior, and that
alterations in oxytonergic signaling are not merely a byproduct of ejac-
ulation (Murphy et al., 1987).
Although considerable attention has been given to the role of OT in
sexual arousal and consummatory sexual behaviors in males, the role
of OT in sexual motivation has received limited attention (Arletti et al.,
1992, 1990, 1985; Gil et al., 2013). The notion that OT influences sexual
motivation is supported by evidence which indicates that oxytonergic
signaling is modulated by testosterone and estrogen, hormones that
play a significant role in male rat sexual motivation (Attila et al.,
2010). Gonadectomized male rats exhibited reduced OT receptor bind-
ing and reduced expression of OT receptor mRNA in the ventromedial
hypothalamus (VMH) (Bale and Dorsa, 1995; Johnson et al., 1991,
1989; Tribollet et al., 1990). Administration of testosterone (Bale and
Dorsa, 1995; Johnson et al., 1991, 1989; Tribollet et al., 1990), or a com-
bination of estradiol and 5-α dihydrotestosterone (DHT) (Johnson et al.,
1991), reversed the effects of gonadectomy on OT receptor binding and
levels of OT receptor mRNA. Notably, in vitro, estradiol has been shown
to modulate the release of OT from hypothalamic neurons (Wang et al.,
1995). Additionally, the aromatase enzyme (El-Emam Dief et al., 2013),
G-protein coupled receptor 30 (GPR30) (Hazell et al., 2009; Sakamoto et
al., 2007) and estrogen receptor β mRNA (Hrabovszky et al., 1998;
Laflamme et al., 1998; Simonian and Herbison, 1997), colocalize with
OT, which may explain why levels of OT receptor mRNA and OT receptor
binding in male rats are influenced by alterations in estrogen signaling
alone (Bale and Dorsa, 1995; Johnson et al., 1991; Tribollet et al., 1990).
Given the relationship between OT and gonadal hormones, as well as
the role that OT plays in consummatory sexual behaviors in male rats, it
is reasonable to hypothesize that administration of an OT receptor an-
tagonist would reduce sexual motivation, as well as consummatory sex-
ual behaviors. However, characterizing the role of oxytocin in sexual
motivation as either the latencies to achieve first mount or intromission,
or the post-ejaculatory interval, does not fully capture the motivational
aspects of sexual behavior because these outcomes do not occur inde-
pendent of changes in consummatory behaviors (Everitt, 1990). With
this in mind, it is advantageous to examine the influence of OT on the
motivational aspects of male rat sexual behavior independent of con-
summatory behaviors. Therefore, it was predicted that administration
of an oxytocin receptor antagonist would reduce the amount of time
male rats would spend adjacent to a sexually receptive female rat that
could not be physically contacted.
2. Methods
2.1. Animals
All procedures were approved by the Franklin and Marshall College
Institutional Animal Care and Use Committee in accordance with the
Guide for the Care and Use of Laboratory Animals (2011). Thirty behav-
iorally naive test (N = 25) and stimulus (N = 5) male Long-Evans rats
were obtained from a breeding colony in the animal care facility at
Franklin and Marshall College. No more than three test male rats were
used from each litter. Each litter was comprised of both drug and vehicle
treated rats. Male rats were between 121 and 126 days of age on the day
of sexual motivation testing. Ovariectomized stimulus female Long-
Evans rats (N = 24), which were between 118 and 120 days of age on
the day of sexual motivation testing, were purchased from Harlan Lab-
oratories (Indianapolis, IN).
All of the test and stimulus rats were group housed by sex and pro-
vided unrestricted access to food and water in polycarbonate cages that
were lined with wood chip bedding. Rats were maintained in a temper-
ature-controlled environment on a reverse 12:12 light-dark schedule
(lights on at 10:00 pm). All behavioral testing was conducted at least
2 h after the beginning of the dark cycle.
2.2. Drug and hormone treatments
Sexual receptivity in stimulus female rats was induced by intramus-
cular injections of estrodial benzoate (10 μg; Custom Prescriptions of
Lancaster, Lancaster, PA) and progesterone (500 μg; Custom Prescrip-
tions of Lancaster) that were delivered 48 h and 4–6 h, respectively,
prior to either sexual experience or the sexual motivation test. Both hor-
mones were suspended in a 0.1 mL vehicle solution that consisted of
sesame oil and benzyl alcohol (3%; Custom Prescriptions of Lancaster).
A timeline depicting the behavioral and pharmacological procedures
conducted with test male rats is shown in Fig. 1. Test male rats received
a single injection into the intraperitoneal cavity of either an OT receptor
antagonist, L368.899 hydrochloride (1 mg/kg; Santa Cruz Biotechnology,
Dallas, TX), or vehicle (sterile water; 1 mL/kg) 40 min prior to the begin-
ning of sexual motivation testing (Boccia et al., 2007). The non-peptide OT
receptor antagonist L368,899 was found in both the cerebrospinal fluid
and hypothalamus of female rhesus monkeys at this dosage and time
point following peripheral administration, an effect that corresponded
with a reduction in sexual behavior (Boccia et al., 2007). In addition,
administration of L368,899 directly into the amygdala has been shown
to counteract the anti-aggressive effects of OT in male rats (Calcagnoli et
al., 2015b) and intraperitoneal administration of L368.899 attenuates
social reward in wild-type male mice (Wei et al., 2015).
2.3. Sexual behavior prescreen
Test male rats were individually acclimated to temporary home cages
on three separate 10-min sessions in a behavioral testing room that was
illuminated by red light with a standard house fan on in a corner of the
room to minimize background noise (Hawley et al., 2011). Prior to the be-
ginning of sexual motivation testing, all test male rats were given three
separate 45-minute sexual experiences with a novel sexually receptive
stimulus female rat in the temporary home cage. Each experience was
separated by one week. The frequencies of mounts, intromissions, and
ejaculations were scored by a trained observer and recorded by a video
camera. Test male rats that did not exhibit at least one mount on any of
the three prescreen tests were removed from the study (N = 1). In addi-
tion to confirming sexual activity in test male rats, stimulus male rats
were given two sexual experiences in temporary home cages prior to
the beginning of sexual motivation testing to confirm sexual activity. All
five stimulus male rats exhibited at least one ejaculation on the first expe-
rience (40 min) and at least two ejaculations on the final experience (1 h).
2.4. Sexual motivation and consummatory sexual behavior testing
As indicated in Fig. 2, the sexual incentive motivation arena
consisted of a primary arena (100 cm × 50 cm × 40 cm) with outer
walls that were constructed of clear Plexiglas. Two internal walls of
black Plexiglas created three equal sized chambers. Each of the internal
walls had an opening in one of the lower corners (10 cm × 10 cm) that
was placed opposite of the opening on the other internal wall and oppo-
site to the smaller stimulus rat chamber to which it provided access.
Stimulus rat chambers (25 cm × 15 cm × 40 cm) were comprised of
three sides constructed of clear Plexiglas and a fourth side constructed
of perforated metal (0.1 cm wide). The side of the stimulus rat chambers
Fig. 1. Experimental timeline of all behavioral and pharmacological treatments involving
test male rats.
 D.S. Blitzer et al. / Hormones and Behavior 94 (2017) 33–39
Fig. 2. Sexual motivation testing arena with walls 40 cm in height. Gray lines =
transparent walls; black lines = opaque walls; dotted lines = perforated metal divider
separating smaller stimulus chamber from end chambers, which were accessible to test
male rats. Superimposed upward diagonal lines in end chambers indicate stimulus rat
approach zones characterized by video tracking software.
constructed of perforated metal faced against a larger opening (23 cm
× 25 cm) that was cut into the center of each of the two larger end
chambers of the primary arena. Consequently, although physical con-
tact was not permitted, the perforated partition allowed for transmis-
sion of visual, olfactory, and auditory cues.
Acclimation to the testing arena and sexual motivation testing were
conducted in the same room as the sexual behavior prescreens, under
red lighting and with a standard house fan on in order to minimize
background noise. Behavior in the arena during the tests was recorded
by an overhead camera. During the acclimation test, which occurred
two to three weeks prior to sexual motivation testing, test male rats
were placed in the center chamber of the primary arena and were
allowed to freely explore all three larger chambers of the arena for
20 min without the presence of stimulus rats. The acclimation period
served to both familiarize rats to the arena and to establish an initial
preference for the larger end chambers. The amount of time spent in
each of the larger end chambers indicated each rat's natural preference.
On the day of sexual motivation testing, a novel sexually receptive
stimulus female rat was placed in one stimulus chamber and an unfa-
miliar sexually active stimulus male rat was placed in the other stimulus
chamber, prior to placement of the test male into the center chamber.
Importantly, stimulus males were from different litters than the test
male rats they were paired with. Upon placement into the center cham-
ber, each test male rat was allowed to freely explore all three larger
chambers of the primary arena for 20 min. The placement of stimulus
rats was contingent upon each test rat's initial preference for the end
chambers during acclimation, such that the stimulus female rat was
placed in the smaller chamber adjacent to the larger end chamber of
the primary arena that was not preferred during the acclimation test.
The three chambers of the primary arena and the smaller stimulus rat
chambers were cleaned thoroughly between trials with 10% ethanol.
Recorded video files were played back and scored by an observer
blind to treatment conditions who used Ethovision 3.0 (Noldus Infor-
mation Technology Inc., Leesburg, VA) to record the time spent in the
end chambers and approach zones. Stimulus rat approach zones,
which provided a more refined analysis of sexual motivation (Agmo,
2003a), were indicated as the halves of the larger end chambers that
were closest to the stimulus rat chambers (see Fig. 1). Greater time
spent in the entire end chamber adjacent to the female, as well as
35
more time spent in the female approach zone, were indicative of higher
levels of sexual motivation. A greater frequency of approach zone visits
was indicative of increased activity. Longer latencies to first enter the
larger two end chambers were indicative of reduced exploratory
behavior.
Immediately after sexual motivation testing, all test male rats were
given sexual experience for 40 min with the same sexually receptive
stimulus female that was used to test sexual motivation. Consummatory
sexual behavior testing, which was recorded by a video camera and
scored by a trained observer that was blind to the conditions, was con-
ducted in the same testing room and in the same temporary home cage
in which the sexual prescreens occurred. The frequencies of mounts and
intromissions, as well as the latencies to achieve first mount, intromis-
sion, ejaculation and the post-ejaculatory interval (latency to achieve
first intromission after first ejaculation) were examined for the first cop-
ulatory series, which consisted of all behaviors prior to the first ejacula-
tion. The total number of ejaculations over the course of the entire 40-
min test was also recorded. One vehicle treated rat that did not exhibit
any mounting behavior during the consummatory sexual behavior test,
but exhibited mounting behavior and intromissions on all three sexual
prescreens, as well as at least one ejaculation on the each of the final
two prescreens, was removed from the study.
2.5. Statistical analyses
One-sample t-tests were conducted to examine preference for the
male and female associated end chambers relative to chance (400 s).
The total time spent in the entire end chamber adjacent to the female
and the male, the time spent in the male and female approach zones,
the latencies to first enter the male and female end chambers, and the
frequency of approach zone entries, were examined by independent
samples t-tests. Dependent variables for consummatory sexual behav-
iors, which were analyzed by independent samples t-tests, were the fre-
quency of ejaculations over the course of the entire 40-minute test, as
well as the latency to first mount, intromission, and ejaculation, the
total number of mounts and intromissions, and the PEI during the FCS.
All statistical analyses were conducted using the Statistical Package for
the Social Sciences (SPSS). Statistical significant was indicated by p b
0.05. Cohen's d effect sizes of 0.2 were considered small, whereas medi-
um and larger effects sizes were indicated by d = 0.5 and d = 0.8,
respectively.
3. Results
As indicated in Fig. 3A, both vehicle treated rats [t(10) = 7.45,
p b 0.001, d = 2.25] and rats treated with an OT receptor antagonist
[t(11) = 5.20, p b 0.001, d = 1.50] spent a greater amount of time in
the end chamber adjacent to the sexually receptive female relative to
chance (400 s). However, when compared to vehicle treatment, adminis-
tration of an OT receptor antagonist attenuated the amount of time spent
in the end chamber adjacent to the sexually receptive female rat [t(21) =
2.16, p = 0.041, d = 0.90]. Vehicle treated rats [t(10) = −3.48, p = 0.006,
d = 1.05], but not rats treated with an OT receptor antagonist [t(11) =
−1.84, p = 0.093, d = 0.53], spent significantly less time in the male as-
sociated end chamber relative to chance, as illustrated in Fig. 3B. However,
there were no differences between vehicle and drug treated rats in the
amount of time spent in the end chamber adjacent to the sexually expe-
rienced stimulus male rat [t(21) = −1.23, p = 0.233, d = 0.51].
A refined analysis of sexual motivation revealed that rats treated
with an OT receptor antagonist spent less time than vehicle control
rats in the female approach zone, which was the half of the female asso-
ciated end chamber that was closest to the female [t(21) = 2.41, p =
0.025, d = 1.01; Fig. 4A]. However, there were no differences between
vehicle and drug treated rats in the amount of time spent in the ap-
proach zone of the stimulus male rat [t(21) = − 0.54, p = 0.598, d =
0.23; Fig. 4B].
36 D.S. Blitzer et al. / Hormones and Behavior 94 (2017) 33–39

900

Total Time in Female

Approach Zone (sec)
750 *
600

450

300

150

0
A Vehicle L-368,899
900

Approach Zone (sec)

Total Time in Male
750

600

450

300

150

0
B Vehicle L-368,899

Fig. 4. Sexual motivation in male rats as a function of drug treatment. Mean (A) total time
spent in female approach zone and (B) mean total time spent in male approach zone.
Sec = seconds; *p b 0.05 vs. vehicle; Error bars represent ± SEM.

Fig. 3. Sexual motivation in male rats as a function of drug treatment. Mean (A) total time
spent in female associated end chamber and (B) mean total time spent in male associated
end chamber. Sec = seconds; *p b 0.05 vs. vehicle; #p b 0.01 vs. chance (dotted line). Error
bars represent ±1 SEM.

45
As expected, administration of an OT receptor antagonist did not af-
Latency to 1st Enter the
Female Associated End

40
fect the latency to first visit the female [t(21) = −0.26, p = 0.796 d =
0.26; see Fig. 5A] or the male [t(21) = −0.93, p = 0.364, d = 0.39; see 35
Chamber (sec)

Fig. 5B] associated end chambers. In addition, there were no differences 30

between rats treated with an OT receptor antagonist and vehicle treated 25
rats in the total number of approach zone entries [t(21) = 0.86, p = 20
0.396, d = 0.36; data not shown]. 15
Unexpectedly, there were no differences between rats treated with 10
an OT receptor antagonist and vehicle treated rats in the latency to
5
achieve first mount [t(21) = 0.07, p = 0.945, d = 0.03], intromission
[t(21) = 0.34, p = 0.741, d = 0.14], and ejaculation [t(11.22) = 1.22, 0
p = 0.247, d = 0.51]. In addition, there was no effect of the OT receptor A Vehicle L-368,899
antagonist on the PEI [t(19) = − 1.51, p = 0.147, d = 0.63], the fre-
quency of mounts [t(16.92) = 0.40, p = 0.699, d = 0.17] and intromis- 45
Latency to 1st Enter the

sions [t(21) = 0.04, p = 971, d = 0.02] during the first copulatory 40

Male Associated End

series, and the frequency of ejaculation across the entire 40 min of the 35
Chamber (sec)

test [t(21) = 0.31, p = 0.763, d = 0.13]. 30

25
4. Discussion 20
15
When allowed to freely explore an arena that did not permit physi-
cal contact between a stimulus male and a stimulus female rat, both ve-
10
hicle and OT receptor antagonist treated rats spent more time with the 5
female than would be expected relative to chance. Importantly, the 0
amount of time spent with the female was reduced in test male rats B Vehicle L-368,899
treated with an OT receptor antagonist relative to vehicle treated rats,
which indicated decreased sexual motivation. However, administration Fig. 5. Initial exploration of sexual motivation testing arena in male rats as a function of
of an OT receptor antagonist did not reduce the amount of time spent in drug treatment. Mean (A) latency to first enter the female and (B) the male associated
the vicinity of the stimulus male rat, which suggests that social end chambers. Sec = seconds. Error bars represent ±1 SEM.
 D.S. Blitzer et al. / Hormones and Behavior 94 (2017) 33–39
exploration was not affected by attenuating oxytonergic signaling. An-
tagonism of OT receptors did not impact initial exploratory behaviors
or overall activity as indicated by a lack of difference between condi-
tions in the latencies to first enter the male and female associated end
chambers and by a similar frequency of approach zone entries, respec-
tively. Unexpectedly, there were no differences between drug and vehi-
cle treated rats in consummatory sexual behaviors, which were
examined immediately after the sexual motivation test. Results of the
study indicate that sexually experienced male rats exhibit a preference
for sexually receptive female rats and that OT signaling plays an impor-
tant role in sexual motivation in male rats.
When given the choice between a sexually receptive stimulus fe-
male or a stimulus male, or when given the choice between a sexually
receptive stimulus female or a non-receptive stimulus female, sexually
naïve test male rats prefer the sexually receptive female, as indicated
by an increased time spent adjacent to the sexually receptive female
(Agmo, 2003a, 2003b; Merkx, 1983). However, other studies have
shown that, unlike sexually experienced male rats, sexually naïve
male rats show no preference when given the choice between a sexually
receptive female rat and a sexually active male rat (Vega Matuszczyk et
al., 1994). In the present study, test male rats were given sexual experi-
ence on three separate occasions prior to the sexual motivation test. Re-
gardless of drug treatment, both groups of rats spent more time with the
sexually receptive female rat relative to chance, which supports past re-
search showing that sexually experienced male rats exhibit a preference
for a sexually receptive female (Vega Matuszczyk et al., 1994).
Results from the current study indicated that administration of an
OT receptor antagonist reduced sexual motivation, when operational-
ized as a decrease in the time spent in the proximity of a sexually recep-
tive female. Consistent with the results from the current study,
alterations in oxytonergic signaling have been shown to impact indica-
tors of sexual motivation in paradigms that permit physical contact be-
tween the test male and stimulus female rat. Specifically, either central
or intraperitoneal administration of OT reduced the post-ejaculatory in-
terval (Arletti et al., 1992, 1990, 1985; Gil et al., 2011) and intra-MPOA
injections of an OT receptor antagonist decreased anogenital investiga-
tion (Gil et al., 2011). Collectively, these results suggest that OT plays a
role in sexual motivation whether or not physical contact is permitted.
However, it is worth noting that intraperitoneal administration of OT
to male rats was unable to restore fluoxetine induced reductions in sex-
ual motivation, which was operationalized by the frequency of level
changes in a bi-level testing arena prior to the introduction of a sexually
receptive female (Cantor et al., 1999). Although the low dose of OT used
was effective at enhancing aspects of consummatory sexual behavior in
other studies (Arletti et al., 1985), it may be the case that somewhat
higher doses, which do not impact locomotion or cause sedation
(Uvnäs-Moberg et al., 1994), are required to restore reductions in sexual
motivation in male rats treated with drugs that enhance serotonergic
activity.
Contrary to the results indicated above that OT in the MPOA is in-
volved in sexual motivation (Gil et al., 2011), past research indicated
that lesions of the MPOA do not affect male rat sexual motivation, as
measured by lever pressing behavior to a conditioned stimulus in
order to gain access to a sexually receptive female (Everitt and Stacey,
1987). However, once exposed to a receptive female, male rats with
MPOA lesions were less likely to mount, intromit, and ejaculate, which
indicates that the MPOA is important for consummatory sexual behav-
iors. Although these findings indicate that the MPOA does not play a sig-
nificant role in male rat sexual motivation, lever pressing to gain access
to a stimulus female rat is significantly reduced by bilateral excitotoxic
lesions of the basolateral amygdala (Everitt et al., 1989). Conversely, le-
sions of the basolateral amygdala do not affect consummatory sexual
behavior, such as the latency to ejaculation. At face value, these results
indicate a dissociation between neural systems in the control of male
rat sexual behavior. However, more recent evidence indicates that the
MPOA plays a role in the sexually motivated behaviors of male rats
37
when characterized as the amount of time spent adjacent to a sexually
receptive stimulus female rat that cannot not be physically contacted
(Hurtazo et al., 2008), which was the case for the current study. Specif-
ically, temporary neural inactivation of the MPOA with lidocaine re-
duced the amount of time male rats spent in the vicinity of a sexually
receptive female rat (Hurtazo et al., 2008). With these results in mind,
it is interesting to consider the possibility that OT receptors in the
basolateral amygdala (Elands et al., 1988) and MPOA (Gil et al., 2013,
2011) may modulate different types of sexually motivated behaviors.
Previous evidence indicates that administration of OT peripherally
or directly into the substantia nigra decreased activity of rats in an
open field (Angioni et al., 2016). Alternatively, locomotor activity in-
creased following direct administration into the substantia nigra of an
OT receptor antagonist. Despite these findings, rats in the current
study treated with an OT receptor antagonist were no different than ve-
hicle treated rats in either overall levels of activity, as indicated by the
total number of approach zone entries, or general exploratory behavior,
as indicated by the latency to first visit the male and female associated
end chambers. In addition, there were no differences between drug
and vehicle treated rats in the amount of time spent in the male associ-
ated end chamber, which indicated that the drug's effect on sexual mo-
tivation was not due to a decrease in social motivation, as posited
elsewhere (Hurtazo et al., 2008). In fact, only vehicle treated rats, and
not rats treated with the OT receptor antagonist, spent significantly
less time in the male associated chamber relative to chance. Collectively,
these results indicate that the reduction in sexual motivation exhibited
by rats treated with an OT receptor antagonist was not affected by alter-
ations in activity, exploratory behavior, or sociality.
There were no significant differences in consummatory sexual be-
haviors between drug and vehicle treated rats in the current study, an
effect consistent with a previous study which found that genetic dele-
tion of the OT gene during development does not impact the consum-
matory sexual behaviors of male mice (Lazzari et al., 2013). However,
in adult male rats, central administration of OT receptor antagonists
have been shown to attenuate consummatory sexual behaviors, such
as the frequency of mounts and intromissions, as well as the likelihood
of ejaculation and the latency to ejaculation (Argiolas et al., 1988; Arletti
et al., 1992; Gil et al., 2011). Although previous evidence indicates the
dose of the drug used in the current study readily crosses the blood
brain barrier in non-human primates (Boccia et al., 2007), we cannot
rule out the possibility that either central administration of the OT an-
tagonist or a higher dose of the drug is necessary to affect the consum-
matory sexual behaviors of male rats.
However, it is also important to note that test male rats in the cur-
rent study were given a 20-minute sexual motivation test immediately
prior to the consummatory sexual behavior test. Previous research indi-
cates that a brief 10–20 minute exposure to a sexually receptive female
placed behind a barrier that prevents physical contact raises levels of
testosterone in both male mice and rats (Amstislavskaya and Popova,
2004; Bonilla-Jaime et al., 2006) and facilitates copulatory behavior in
male rats (de Jonge et al., 1992). In addition to the reflexive release of
testosterone that occurs in males following exposure to sexually recep-
tive females, endogenous levels of OT tend to rise in the presence of a
sexually receptive female placed behind a perforated wall (Waldherr
and Neumann, 2007). Conceivably, enhanced neuroendocrine respon-
siveness as a result of exposure to sexual stimuli may have overcome
the effects of the OT receptor antagonist on consummatory sexual
behavior.
The results of the current study have important implications for cli-
nicians working with those suffering from premature ejaculation, which
is one of the most common sexual dysfunctions in men (Laumann et al.,
1999). Popular pharmacological interventions for treating premature
ejaculation include the use of short-term selective serotonin reuptake
inhibitors (SSRIs) (Giuliano and Clèment, 2012). However, patients
who use SSRIs often report lower sexual desire (Rosen et al., 1999). In
light of promising preclinical findings in rats, which indicated that OT
38 D.S. Blitzer et al. / Hormones and Behavior 94 (2017) 33–39
receptor antagonists lengthen the latency to ejaculate (Argiolas et al.,
1989; Clément et al., 2013), reductions in OT signaling may be a viable
option for the treatment of premature ejaculation in men. Treatment
with an OT receptor antagonist resulted in slightly longer ejaculation la-
tencies relative to placebo, although the effect was not significant
(Shinghal et al., 2013). As suggested elsewhere, the potential of OT an-
tagonists to treat premature ejaculation may be contingent upon how
well they cross the blood brain barrier (McMahon, 2016). Importantly,
the results of the current study indicate that treatment with OT receptor
antagonists that appear to cross the blood barrier (Boccia et al., 2007)
could have negative side effects on the motivational aspects of sexual
behavior in men, an effect similar to that which occurs following treat-
ment with an SSRI.
Acknowledgments
This research was supported by a Leser Grant (DB) and Faculty Re-
search/Professional Development Funds from the Office of the Provost
at Franklin & Marshall College (WH). The authors gratefully acknowl-
edge Francis “Frank” Koczur and Steven Spadafore for their technical
support, as well as Lillian Basom and Richelle Wagner for their expertise
in animal care.
References
Agmo, A., 2003a. Unconditioned sexual incentive motivation in the male Norway rat
(Rattus norvegicus). J. Comp. Psychol. 117, 3–14.
Agmo, A., 2003b. Lack of opioid or dopaminergic effects on unconditioned sexual incen-
tive motivation in male rats. Behav. Neurosci. 117, 55–68.
Amstislavskaya, T., Popova, N., 2004. Female-induced sexual arousal in male mice and
rats: behavioral and testosterone response. Horm. Behav. 46, 544–550.
Angioni, L., Cocco, C., Ferri, G., Argiolas, A., Melis, M., Sanna, F., 2016. Involvement of nigral
oxytocin in locomotor activity: a behavioral, immunohistochemical and lesion study
in male rats. Horm. Behav. 83, 23–38.
Argiolas, A., 1999. Neuropeptides and sexual behaviour. Neurosci. Biobehav. Rev. 23,
1127–1142 (doi:S0149763499000688 [pii]).
Argiolas, A., Melis, M.R., Vargiu, L., Gessa, G.L., 1987. d(CH2)5Tyr(Me)-[Orn8]vasotocin, a
potent oxytocin antagonist, antagonizes penile erection and yawning induced by
oxytocin and apomorphine, but not by ACTH-(1-24). Eur. J. Pharmacol. 134,
221–224 (doi:0014-2999(87)90168-3 [pii]).
Argiolas, A., Collu, M., Gessa, G.L., Melis, M.R., Serra, G., 1988. The oxytocin antagonist
d(CH2)5Tyr(Me)-Orn8-vasotocin inhibits male copulatory behaviour in rats. Eur.
J. Pharmacol. 149, 389–392.
Argiolas, A., Collu, M., D'Aquila, P., Gessa, G.L., Melis, M.R., Serra, G., 1989. Apomorphine
stimulation of male copulatory behavior is prevented by the oxytocin antagonist
d(CH2)5 Tyr(Me)-Orn8-vasotocin in rats. Pharmacol. Biochem. Behav. 33, 81–83.
Arletti, R., Bazzani, C., Castelli, M., Bertolini, A., 1985. Oxytocin improves male copulatory
performance in rats. Horm. Behav. 19, 14–20 (doi:0018-506X(85)90002-9 [pii]).
Arletti, R., Benelli, A., Bertolini, A., 1990. Sexual behavior of aging male rats is stimulated
by oxytocin. Eur. J. Pharmacol. 179, 377–381.
Arletti, R., Benelli, A., Bertolini, A., 1992. Oxytocin involvement in male and female sexual
behavior. Ann. N. Y. Acad. Sci. 652, 180–193.
Attila, M., Oksala, R., Agmo, A., 2010. Sexual incentive motivation in male rats requires
both androgens and estrogens. Horm. Behav. 58:341–351 (doi:S0018-
506X(09)00197-4 [pii]). http://dx.doi.org/10.1016/j.yhbeh.2009.08.011.
Bale, T.L., Dorsa, D.M., 1995. Regulation of oxytocin receptor messenger ribonucleic acid in
the ventromedial hypothalamus by testosterone and its metabolites. Endocrinology
136:5135–5138. http://dx.doi.org/10.1210/endo.136.11.7588251.
Boccia, M.L., Goursaud, A.P., Bachevalier, J., Anderson, K.D., Pedersen, C.A., 2007. Peripher-
ally administered non-peptide oxytocin antagonist, L368,899, accumulates in limbic
brain areas: a new pharmacological tool for the study of social motivation in non-
human primates. Horm. Behav. 52:344–351 (doi:S0018-506X(07)00130-4 [pii]).
http://dx.doi.org/10.1016/j.yhbeh.2007.05.009.
Bonilla-Jaime, H., Vazquez-Palacios, G., Arteaga-Silva, M., Retana-Marquez, S., 2006. Hor-
monal responses to different sexually related conditions in male rats. Horm. Behav.
49:376–382 (doi:S0018-506X(05)00208-4 [pii]). http://dx.doi.org/10.1016/j.yhbeh.
2005.08.005.
Calcagnoli, F., Kreutzmann, J.C., de Boer, S.F., Althaus, M., Koolhaas, J.M., 2015a. Acute and
repeated intranasal oxytocin administration exerts anti-aggressive and pro-affiliative
effects in male rats. Psychoneuroendocrinology 51:112–121 (doi:S0306-
4530(14)00359-X [pii]). http://dx.doi.org/10.1016/j.syneuen.2014.09.019.
Calcagnoli, F., Stubbendorff, C., Meyer, N., de Boer, S.F., Althaus, M., Koolhaas, J.M., 2015b.
Oxytocin microinjected into the central amygdaloid nuclei exerts anti-aggressive ef-
fects in male rats. Neuropharmacology 90:74–81 (doi:S0028-3908(14)00428-6 [pii]).
http://dx.doi.org/10.1016/j.neuropharm.2014.11.012.
Cantor, J.M., Binik, Y.M., Pfaus, J.G., 1999. Chronic fluoxetine inhibits sexual behavior in
the male rat: reversal with oxytocin. Psychopharmacology 144, 355–362.
Clément, P., Bernabé, J., Compagnie, S., Alexandre, L., McCallum, S., Giuliano, F., 2013. In-
hibition of ejaculation by the non-peptide oxytocin receptor antagonist
GSK557296: a multi-level site of action. Br. J. Pharmacol. 169:1477–1485. http://dx.
doi.org/10.1111/bph.12198.
Elands, J., Beetsma, A., Barberis, C., de Kloet, E.R., 1988. Topography of the oxytocin recep-
tor system in rat brain: an autoradiographical study with a selective radioiodinated
oxytocin antagonist. J. Chem. Neuroanat. 1, 293–302.
El-Emam Dief, A., Caldwell, J.D., Jirikowski, G.F., 2013. Colocalization of p450 aromatase
and oxytocin immunostaining in the rat hypothalamus. Horm. Metab. Res. 45:
273–276. http://dx.doi.org/10.1055/s-0032-1327680.
Everitt, B., 1990. Sexual motivation: a neural and behavioural analysis of the mechanisms
underlying appetitive and copulatory responses of male rats. Neurosci. Biobehav. Rev.
14, 217–232.
Everitt, B.J., Stacey, P., 1987. Studies of instrumental behavior with sexual reinforcement
in male rats (Rattus norvegicus): II. Effects of preoptic area lesions, castration, and tes-
tosterone. J. Comp. Psychol. 101, 407–419.
Everitt, B.J., Cador, M., Robbins, T.W., 1989. Interactions between the amygdala and ven-
tral striatum in stimulus-reward associations: studies using a second-order schedule
of sexual reinforcement. Neuroscience 30, 63–75.
George, J.M., Staples, S., Marks, B.M., 1976. Oxytocin content of microdissected areas of rat
hypothalamus. Endocrinology 98:1430–1433. http://dx.doi.org/10.1210/endo-98-6-
1430.
Gil, M., Bhatt, R., Picotte, K.B., Hull, E.M., 2011. Oxytocin in the medial preoptic area facil-
itates male sexual behavior in the rat. Horm. Behav. 59:435–443 (doi:S0018-
506X(10)00318-1 [pii]). http://dx.doi.org/10.1016/j.yhbeh.2010.12.012.
Gil, M., Bhatt, R., Picotte, K.B., Hull, E.M., 2013. Sexual experience increases oxytocin re-
ceptor gene expression and protein in the medial preoptic area of the male rat.
Psychoneuroendocrinology 38:1688–1697 (doi:S0306-4530(13)00040-1 [pii]).
http://dx.doi.org/10.1016/j.psyneuen.2013.02.002.
Giuliano, F., Clèment, P., 2012. Pharmacology for the treatment of premature ejaculation.
Pharmacol. Rev. 64:621–644. http://dx.doi.org/10.1124/pr.111.004952.
Hawley, W., Grissom, E., Keskitalo, L., Hastings, T., Dohanich, G., 2011. Sexual motivation
and anxiety-like behaviors of male rats after exposure to a trauma followed by situ-
ational reminders. Physiol. Behav. 102:181–187 (doi:S0031-9384(10)00384-7 [pii]).
http://dx.doi.org/10.1016/j.physbeh.2010.10.021.
Hazell, G.G., Yao, S.T., Roper, J.A., Prossnitz, E.R., O'Carroll, A.M., Lolait, S.J., 2009.
Localisation of GPR30, a novel G protein-coupled oestrogen receptor, suggests multi-
ple functions in rodent brain and peripheral tissues. J. Endocrinol. 202:223–236.
http://dx.doi.org/10.1677/JOE-09-0066.
Hillegaart, V., Alster, P., Uvnäs-Moberg, K., Ahlenius, S., 1998. Sexual motivation promotes
oxytocin secretion in male rats. Peptides 19:39–45. http://dx.doi.org/10.1016/S0196-
9781(97)00250-7.
Hrabovszky, E., Kalló, I., Hajszán, T., Shughrue, P.J., Merchenthaler, I., Liposits, Z., 1998. Ex-
pression of estrogen receptor-beta messenger ribonucleic acid in oxytocin and vaso-
pressin neurons of the rat supraoptic and paraventricular nuclei. Endocrinology 139:
2600–2604. http://dx.doi.org/10.1210/endo.139.5.6024.
Hughes, A.M., Everitt, B.J., Lightman, S.L., Todd, K., 1987. Oxytocin in the central nervous
system and sexual behaviour in male rats. Brain Res. 414, 133–137 (doi:0006-
8993(87)91333-3 [pii]).
Hurtazo, H.A., Paredes, R.G., Ågmo, A., 2008. Inactivation of the medial preoptic area/ante-
rior hypothalamus by lidocaine reduces male sexual behavior and sexual incentive
motivation in male rats. Neuroscience 152:331–337. http://dx.doi.org/10.1016/j.
neuroscience.2007.10.063.
Johnson, A.E., Coirini, H., McEwen, B.S., Insel, T.R., 1989. Testosterone modulates oxytocin
binding in the hypothalamus of castrated male rats. Neuroendocrinology 50, 199–203.
Johnson, A.E., Coirini, H., Insel, T.R., McEwen, B.S., 1991. The regulation of oxytocin recep-
tor binding in the ventromedial hypothalamic nucleus by testosterone and its metab-
olites. Endocrinology 128:891–896. http://dx.doi.org/10.1210/endo-128-2-891.
de Jonge, F., Oldenburger, W., Louwerse, A., Van de Poll, N., 1992. Changes in male copu-
latory behavior after sexual exciting stimuli: effects of medial amygdala lesions. Phys-
iol. Behav. 52, 327–332.
Laflamme, N., Nappi, R.E., Drolet, G., Labrie, C., Rivest, S., 1998. Expression and
neuropeptidergic characterization of estrogen receptors (ERalpha and ERbeta)
throughout the rat brain: anatomical evidence of distinct roles of each subtype.
J. Neurobiol. 36:357–378. http://dx.doi.org/10.1002/(SICI)1097-4695(19980905)36:
3b357::AID-NEU5N3.0.CO;2-V (pii).
Laumann, E.O., Paik, A., Rosen, R.C., 1999. Sexual dysfunction in the United States: preva-
lence and predictors. JAMA 281, 537–544.
Lazzari, V.M., Becker, R.O., de Azevedo, M.S., Morris, M., Rigatto, K., Almeida, S., Lucion,
A.B., Giovenardi, M., 2013. Oxytocin modulates social interaction but is not essential
for sexual behavior in male mice. Behav. Brain Res. 244:130–136. http://dx.doi.org/
10.1016/j.bbr.2013.01.025.
Liu, Y.C., Salamone, J.D., Sachs, B.D., 1997. Impaired sexual response after lesions of the
paraventricular nucleus of the hypothalamus in male rats. Behav. Neurosci. 111,
1361–1367.
Lukas, M., Toth, I., Reber, S.O., Slattery, D.A., Veenema, A.H., Neumann, I.D., 2011. The neu-
ropeptide oxytocin facilitates pro-social behavior and prevents social avoidance in
rats and mice. Neuropsychopharmacology 36:2159–2168 (doi:npp201195 [pii]).
http://dx.doi.org/10.1038/npp.2011.95.
Lukas, M., Toth, I., Veenema, A.H., Neumann, I.D., 2013. Oxytocin mediates rodent social
memory within the lateral septum and the medial amygdala depending on the rele-
vance of the social stimulus: male juvenile versus female adult conspecifics.
Psychoneuroendocrinology 38:916–926 (doi:S0306-4530(12)00334-4 [pii]). http://
dx.doi.org/10.1016/j.psyneuen.2012.09.018.
McMahon, C.G., 2016. Emerging and investigational drugs for premature ejaculation.
Transl. Androl. Urol. 5:487–501. http://dx.doi.org/10.21037/tau.2016.04.02.
 D.S. Blitzer et al. / Hormones and Behavior 94 (2017) 33–39
Melis, M.R., Argiolas, A., Gessa, G.L., 1986. Oxytocin-induced penile erection and yawning:
site of action in the brain. Brain Res. 398, 259–265 (doi:0006-8993(86)91485-X
[pii]).
Melis, M.R., Stancampiano, R., Argiolas, A., 1992. Hippocampal oxytocin mediates apo-
morphine-induced penile erection and yawning. Pharmacol. Biochem. Behav. 42,
61–66.
Melis, M.R., Spano, M.S., Succu, S., Argiolas, A., 1999. The oxytocin antagonist
d(CH2)5Tyr(Me)2-Orn8-vasotocin reduces non-contact penile erections in male
rats. Neurosci. Lett. 265, 171–174 (doi:S0304394099002360 [pii]).
Melis, M.R., Succu, S., Sanna, F., Boi, A., Argiolas, A., 2009. Oxytocin injected into the ven-
tral subiculum or the posteromedial cortical nucleus of the amygdala induces penile
erection and increases extracellular dopamine levels in the nucleus accumbens of
male rats. Eur. J. Neurosci. 30:1349–1357 (doi:EJN6912 [pii]). http://dx.doi.org/10.
1111/j.1460-9568.2009.06912.x.
Merkx, J., 1983. Sexual motivation of the male rat during the oestrous cycle of the female
rat. Behav. Brain Res. 7, 229–237.
Murphy, M.R., Seckl, J.R., Burton, S., Checkley, S.A., Lightman, S.L., 1987. Changes in oxyto-
cin and vasopressin secretion during sexual activity in men. J. Clin. Endocrinol. Metab.
65:738–741. http://dx.doi.org/10.1210/jcem-65-4-738.
National Research Council, 2011. Guide for the Care and Use of Laboratory Animals. 8th ed.
National Academies Press, Washington, DC, USA.
Ramos, L., Hicks, C., Caminer, A., Goodwin, J., McGregor, I.S., 2015. Oxytocin and MDMA
(‘ecstasy’) enhance social reward in rats. Psychopharmacology 232:2631–2641.
http://dx.doi.org/10.1007/s00213-015-3899-9.
Rosen, R.C., Lane, R.M., Menza, M., 1999. Effects of SSRIs on sexual function: a critical re-
view. J. Clin. Psychopharmacol. 19, 67–85.
Sakamoto, H., Matsuda, K., Hosokawa, K., Nishi, M., Morris, J.F., Prossnitz, E.R., Kawata, M.,
2007. Expression of G protein-coupled receptor-30, a G protein-coupled membrane
estrogen receptor, in oxytocin neurons of the rat paraventricular and supraoptic nu-
clei. Endocrinology 148:5842–5850. http://dx.doi.org/10.1210/en.2007-0436.
Shinghal, R., Barnes, A., Mahar, K., Stier, B., Giancaterino, L., Condreay, L., Black, L.,
McCallum, S., 2013. Safety and efficacy of epelsiban in the treatment of men with pre-
mature ejaculation: a randomized, double-blind, placebo-controlled, fixed-dose
study. J. Sex. Med. 10, 2506–2517.
39
Simonian, S.X., Herbison, A.E., 1997. Differential expression of estrogen receptor alpha
and beta immunoreactivity by oxytocin neurons of rat paraventricular nucleus.
J. Neuroendocrinol. 9, 803–806.
Stoneham, M.D., Everitt, B.J., Hansen, S., Lightman, S.L., Todd, K., 1985. Oxytocin and sex-
ual behaviour in the male rat and rabbit. J. Endocrinol. 107, 97–106.
Succu, S., Sanna, F., Cocco, C., Melis, T., Boi, A., Ferri, G.L., Argiolas, A., Melis, M.R., 2008.
Oxytocin induces penile erection when injected into the ventral tegmental area of
male rats: role of nitric oxide and cyclic GMP. Eur. J. Neurosci. 28:813–821 (doi:
EJN6385 [pii]). http://dx.doi.org/10.1111/j.1460-9568.2008.06385.x.
Succu, S., Sanna, F., Argiolas, A., Melis, M.R., 2011. Oxytocin injected into the hippocampal
ventral subiculum induces penile erection in male rats by increasing glutamatergic
neurotransmission in the ventral tegmental area. Neuropharmacology 61:181–188
(doi:S0028-3908(11)00143-2 [pii]). http://dx.doi.org/10.1016/j.neuropharm.2011.
03.026.
Tribollet, E., Audigier, S., Dubois-Dauphin, M., Dreifuss, J.J., 1990. Gonadal steroids regulate
oxytocin receptors but not vasopressin receptors in the brain of male and female rats.
An autoradiographical study. Brain Res. 511, 129–140 (doi:0006-8993(90)90232-Z
[pii]).
Uvnäs-Moberg, K., Ahlenius, S., Hillegaart, V., Alster, P., 1994. High doses of oxytocin cause
sedation and low doses cause an anxiolytic-like effect in male rats. Pharmacol.
Biochem. Behav. 49, 101–106.
Vega Matuszczyk, J., Shree Appa, R., Larsson, K., 1994. Age-dependent variations in the
sexual preference of male rats. Physiol. Behav. 55, 827–830 (doi:0031-
9384(94)90067-1 [pii]).
Waldherr, M., Neumann, I.D., 2007. Centrally released oxytocin mediates mating-induced
anxiolysis in male rats. Proc. Natl. Acad. Sci. U. S. A. 104:16681–16684 (doi:
0705860104 [pii]). http://dx.doi.org/10.1073/pnas.0705860104.
Wang, I., Ward, A., Morris, J., 1995. Oestradiol acutely stimulates exocytosis of oxytocin
and vasopressin from dendrites and somata of hypothalamic magnocellular neurons.
Neuroscience 68, 1179–1188.
Wei, D., Lee, D., Cox, C.D., Karsten, C.A., Peñagarikano, O., Geschwind, D.H., Gall, C.M.,
Piomelli, D., 2015. Endocannabinoid signaling mediates oxytocin-driven social re-
ward. Proc. Natl. Acad. Sci. U. S. A. 112:14084–14089. http://dx.doi.org/10.1073/
pnas.1509795112.