At the Cutting Edge
Neuroendocrinology 2018;107:400–416
DOI: 10.1159/000494558
Sexual Behavior: From Hormonal
Regulation to Endocrine Disruption
Sakina Mhaouty-Kodja Lydie Naulé Daphné Capela
CNRS, INSERM, Neuroscience Paris Seine – Institut de Biologie Paris Seine, Sorbonne Université, Paris, France
Keywords
Sexual behavior · Endocrine disruptor · Sex steroids ·
Reproduction · Nervous system
Abstract
Sexual behavior constitutes a chain of behavioral responses
beginning with courtship and leading to copulation. These
responses, which are exhibited in a sexually dimorphic man-
ner by the two partners, are tightly regulated by sex steroid
hormones as early as the perinatal period. Hormonal chang-
es or exposure to exogenous factors exhibiting hormone-
mimetic activities, such as endocrine disrupting compounds
(EDC), can therefore interfere with their expression. Here we
review the experimental studies in rodents performed to ad-
dress the potential effects of exposure to EDC on sexual be-
havior and underlying mechanisms, with particular atten-
tion to molecules with estrogenic and/or anti-androgenic
activities. © 2018 S. Karger AG, Basel
Introduction
Sexual reproduction in vertebrates requires sexually
differentiated behaviors to ensure attraction between
partners and mating. These behaviors are a key factor in
© 2018 S. Karger AG, Basel
E-Mail karger@karger.com
www.karger.com/nen
Received: July 24, 2018
Accepted after revision: October 16, 2018
Published online: October 16, 2018
the success of sexual reproduction, on which species sur-
vival depends. Males and females adopt different behav-
iors and postures during attraction and mating. Male ro-
dents readily express sexual behavior whenever a recep-
tive female is present. During the precopulatory or
appetitive phase, males engage in chemoinvestigation,
displaying an olfactory preference for receptive females.
Males generate ultrasonic vocalizations in response to fe-
males or their odors [1–3]. These vocalizations have char-
acteristics in common with the songs of songbirds [4].
The emission of courtship vocalizations conveys infor-
mation about the male’s motivational state and helps to
attract the female partner. During the copulatory or con-
summatory phase, males mount, thrust, intromit. In male
mice, mating ends with ejaculation, whereas male rats
reach satiety after several ejaculations and do not then
copulate again for 1–3 days [5].
In female rodents, receptivity is constrained by
threshold levels of estradiol (E2) following progesterone
secretion, and is, therefore, limited to the estrous phase
of the cycle corresponding to the ovulation period. The
female also participates actively in mating, through
three phases of sexual behavior: attractivity, proceptiv-
ity, and receptivity [6]. During attractivity, the female
stimulates male behavior by emitting pheromones. The
female then adopts several proceptive behaviors (hops,
darts, solicitations) in response to stimuli from the male
Sakina Mhaouty-Kodja
CNRS UMR 8246, INSERM U1130, Sorbonne Université
7, quai St Bernard, Bât A 3ème étage
FR–75005 Paris (France)
E-Mail sakina.mhaouty-kodja @ sorbonne-universite.fr
[7]. Finally, during the copulatory phase, the female
adopts a receptive posture called lordosis while ap-
proached from behind for insemination by the courting
male [8].
Sexual behavior is tightly controlled by finely tuned
neural processes, which begin during development and
are tightly regulated by gonadal hormones. These pro-
cesses may, therefore, be highly sensitive to exposure to
endocrine disrupting compounds (EDC), defined by the
WHO and the Endocrine Society as exogenous chemical
substances or mixture of substances that alter the func-
tions of the endocrine system. Exposure to EDC has been
reported to alter several reproductive functions, includ-
ing organ development, germ cell production, pubertal
timing and other physiological processes required for
fertility, as documented by both experimental and epide-
miological studies [9]. Several studies have addressed the
effects of exposure to EDC on mating behavior. Here, we
review the experimental studies in rodents performed to
address the potential effects of exposure to EDC on sex-
ual behavior. We focus on the underlying molecular and
neural processes affected by such exposure, paying par-
ticular interest to molecules with estrogenic and/or anti-
androgenic activities. A recent review by Gore et al. [10]
provided the general ethological background to sexual
selection and reproductive competence in animal species
and their sensitivity to EDC.
Hormonal Regulation of Sexual Behavior and
Underlying Sexual Dimorphisms
Critical Periods of Hormonal Regulation
Perinatal Period
Since the pioneering work of Phoenix et al. [11], it has
become clear that gonadal hormones play a key role in the
sexual differentiation of mating behavior. This process
begins early, in the perinatal (late gestational and early
neonatal) period. In males, testosterone released from the
fetal and neonatal testes permanently potentiates male
(masculinization) behavioral and anatomic characteris-
tics whilst inhibiting female (defeminization) character-
istics in the neural circuitry underlying sexual behavior.
This circuitry is stimulated by pheromonal cues emitted
by receptive females and transmitted from the main ol-
factory epithelium and vomeronasal organ to the main
and accessory olfactory bulbs, respectively, and then to
the chemosensory responsive nuclei in the medial amyg-
dala, the bed nucleus of the stria terminalis, and the me-
dial preoptic area, where they are processed in behavioral
Endocrine Disruption of Sexual Behavior
responses. Projections are sent from the hypothalamic
paraventricular nucleus to the spinal centers that pro-
mote penile erection and ejaculation, including the spinal
nucleus of the bulbocavernosus, and the gastrin-releasing
peptide system.
Perinatal testosterone secretion has organizational ef-
fects, resulting in structural, neurochemical and molecu-
lar differences in circuitry between the sexes. Differences
in cell number and morphology or fiber density between
the sexes have frequently been described for the medial
amygdala, the bed nucleus of the stria terminalis, and the
medial preoptic area [12]. For instance, the rat sexually
dimorphic nucleus (SDN) and corresponding cluster of
calbindin-immunoreactive neurons, both located in the
medial preoptic area, contain more cells in males than in
females [13, 14]. Conversely, neurons expressing kiss-
peptin and tyrosine hydroxylase in the anteroventral
periventricular (AVPV) nucleus, a subdivision of the
preoptic area involved in the ovulatory surge of LH, are
more numerous in females than in males [15–17]. The
regulation of these neuronal populations, or of other sex-
ually dimorphic features, by perinatal testosterone can be
mimicked by E2, because gonadal testosterone is aroma-
tized into neural E2 by the aromatase cytochrome P450.
At the molecular level, testosterone and its neural me-
tabolite E2 also trigger differences in gene expression be-
tween the sexes, through long-lasting changes including
epigenetic modifications [18].
The neural circuitry underlying behavior in females
also includes the olfactory bulb, which transmits signals
to the medial amygdala and then to the bed nucleus of
the stria terminalis, the preoptic area, and the ventrome-
dial hypothalamus, the principal facilitatory system for
lordosis behavior. Systems inhibiting lordosis and in-
volving the lateral septum, the preoptic area, and the ar-
cuate nucleus have also been reported. These nuclei also
receive projections from the medial amygdala and the
bed nucleus of the stria terminalis. These inhibitory and
facilitatory systems send projections to the periaque-
ductal gray regions through the ventromedial hypothal-
amus and lateral septum, respectively. These projections
relay the information to spinal motoneurons innervat-
ing the axial muscles involved in the lordosis posture
[19]. During the prenatal and early postnatal periods,
these neural structures are protected from the masculin-
izing effects of sex steroids by the presence of
α-fetoprotein, which selectively sequesters E2 derived
from the mother and male siblings since the fetal ovaries
are inactive [20].
Neuroendocrinology 2018;107:400–416 401
DOI: 10.1159/000494558
 Prepubertal/Pubertal Period
The pubertal period is characterized by the central ac-
tivation of pulsatile GnRH secretion, which stimulates
the pituitary gonadotropin secretion required for sexual
maturation and fertility. During this period, testosterone
also exerts long-term effects on behavior to ensure the
maturation of processes initiated during the perinatal pe-
riod. Indeed, the prepubertal castration of male hamsters
has been shown to decrease sexual behavior (fewer
mountings and intromissions and longer times to ejacu-
lation) and to shorten the time to lordosis relative to that
for males castrated after puberty [21]. Neural processes,
such as cell proliferation, in sexually dimorphic regions
are influenced by gonadal hormones during the peripu-
bertal period [22, 23].
In females, α-fetoprotein levels decrease after birth
and the ovaries begin releasing E2 on postnatal day (PND)
7. Postnatal/prepubertal E2 secretion plays an active role
in the feminization of sexual behavior. Indeed, E2 admin-
istration between PND15 and PND25 in aromatase
knockout mice, which normally have highly impaired
lordosis behavior, partially restores this behavior [24].
During this postnatal period, ovarian E2 is also essential
for the establishment of the LH surge, which is synchro-
nized with the receptivity period. In particular, prepuber-
tal E2 activates an increase in kisspeptin expression in the
rostral periventricular area of the third ventricle [25–27].
Adulthood
In adult males, gonadal testosterone acts on the male
neural circuitry to stimulate sexual behavior. This activa-
tional effect of testosterone is transient by comparison to
the permanent organizational changes induced during
the developmental and pubertal periods. Male sexual
stimulation is reduced or inhibited by castration but can
be restored by hormonal supplementation. Testosterone
and its neural metabolite E2 regulate the signaling path-
ways of neurotransmitters and neuropeptides playing an
important role in displays of sexual behavior, such as oxy-
tocin, dopamine, and glutamate.
Cyclic females mate only during the estrous phase and
are sexually inactive during the rest of the cycle [28]. The
preovulatory surge of E2, which occurs during the proes-
trus phase, triggers not only an ovulatory surge of LH, but
also the expression of progesterone receptors (PR) in the
ventromedial hypothalamus [29, 30]. Progesterone re-
lease under the control of LH then induces female recep-
tivity, which is perfectly synchronized with ovulation in
such species [28]. The increase in E2 levels also relieves
constraints exerted by the inhibitory system through sup-
402 Neuroendocrinology 2018;107:400–416
DOI: 10.1159/000494558
pression of the inhibition exerted by the lateral septum
and inactivation of the β-endorphin system in the preop-
tic area. Several neuropeptides and neurotransmitters
present in the ventromedial hypothalamus display differ-
ences between the sexes. For example, estrogen receptors
(ER), PR GABA, and enkephalin are all more abundant
in females than in males and are known to promote fe-
male sexual behavior [31].
Mechanisms Underlying the Hormonal Regulation of
Sexual Behavior
Sex steroids regulate sexual behavior principally
through nuclear superfamily receptors. As the detailed
mechanisms underlying the expression of sexual behav-
ior have been largely reviewed, this paragraph summa-
rizes very briefly the genetic studies investigating the rel-
ative contribution of androgen (AR) and ER. Indeed, the
involvement of each of these receptors in this regulation
has been studied by ubiquitous gene invalidations. In
males, data from the testicular feminization mutation and
global AR knockout models, both of which result in a
feminine phenotype, have suggested that this receptor
plays some kind of role in the expression of sexual behav-
ior in rats and mice [32]. Global ERα knockout mice are
infertile and have impaired sexual behavior [33]. Global
ERβ knockout males have normal sexual behavior [34],
but mutant males derived from an ERβ knockout mouse
line completely devoid of ERβ transcripts [35] are infer-
tile and display a mild impairment of sexual behavior
[36]. In the global ERα and ERβ knockout models, fe-
males are infertile, with much lower frequencies of lor­
dosis and proceptive behaviors than wild-type females
[36, 37].
The neural role of these receptors was dissected more
precisely by conditional gene invalidation, which has the
advantage of preserving peripheral receptor expression
and not interfering with gonadal and genital tract effects.
In males, the estrogenic and androgenic pathways seem
to play complementary roles in the organization and ac-
tivation of the neural circuitry underlying sexual behav-
ior. The perinatal masculinization and defeminization of
brain structures involve the estrogenic pathway, whereas
the neural AR is involved principally in the postnatal or-
ganization of spinal nuclei; however, both pathways seem
to be involved in the adult activation of sexual behavior
[32].
In females, neural ERβ invalidation delays the onset of
puberty but has no effect on sexual behavior and fertility
in either naïve or sexually experienced mutants [38]. This
suggests that the ovary may be the primary site of ERβ ac-
Mhaouty-Kodja/Naulé/Capela
Table 1. The analyzed EDC presented by hormonal activity, uses and reference doses established by agencies when available

Source or use Reference doses

Estrogenic compounds
Bisphenol A (BPA) In polycarbonate plastic used in the manufacture of TDI = 0.004 mg/kg/day (EFSA, 2015)
food containers, dental resins, thermal papers TDI = 0.05 mg/kg/day (US EPA, 2012)
Diethylstilbestrol (DES) Used in cases of preterm labor. Banned in 1971 in the
USA and since 1975 in several European countries
Ethinyl estradiol (EE) Oral contraceptive treatment
Methoxychlor (MXC) Organochlorine insecticide banned since 2002 in EU ADI: 0.1 mg/kg/day (EU Pesticides
and 2003 in the USA. Still present in the environment Database)
and used for applications such as mosquito and malaria
control in developing countries
Nonylphenol (NP) Nonionic surfactant used in industrial, agricultural and TDI: 0.005 mg/kg/day (Danish EPA,
domestic applications such as soap, cosmetics, paints, 2000)
herbicides and pesticides, or plastic fabrication
Phytoestrogens
Coumestrol Present in plants (Alfalfa, soybeans…)
Ferutinin Present in roots of Ferula plants
Isoflavones Present in plants (lupin, soybeans, fava beans…) LOAEL of 35 mg/kg/day for isoflavones
(daidzein, genistein) with safety factor of 300 (Anses 2015)
Resveratrol (RVT) Present in grapes, blueberries, raspberries…
Anti-androgenic compounds
Phthalates
Dibutyl phthalate (DBP) Used in the manufacture of PVC plastics, adhesives, TDI: 0.01 mg/kg/day (EFSA, 2005)
inks DNEL: 0.0067 mg/kg/day (ECHA 2017)
Di-(2-ethylhexyl) adipate Used as a substitute for DEHP (marginal use): PVC,
(DEHA) food plastic, cosmetics
Di-(2-ethylexyl)phthalate Used in PVC, food containers, medical products, TDI: 0.050 mg/kg/day (EFSA, 2005)
(DEHP) automobile… NOAEL: 4.8 mg/kg/day, DNEL: 0.034 mg/
kg/day (ECHA 2017)
Diisononyl phthalate Used as a replacement for DEHP: PVC, paint, inks, Sum DIDP/DINP: 0.15 mg/kg/day
(DINP) toys, plastic, soles, automobile (EFSA, 2005)
Vinclozolin Fungicide widely used on fruits and vegetables to avoid ADI = 0.01 mg/kg/day (WHO, 1998)
rot and mildew NOAEL = 1.2 mg/kg/day (US EPA, 2003)
ADI = 0.005 mg/kg/day (EU Pesticides
Database)

ADI, acceptable daily intake dose; DNEL, derived no-effect level; LOAEL, lowest observed adverse effect level; NOAEL, no-observed
effect level; TDI, tolerable daily intake dose.

tion in these processes in adults. Further studies are re-
quired to address the behavioral phenotype of neural ERα
knockout mice, but this receptor seems to play a major
role in the E2-dependent regulation of reproductive pro-
cesses, as demonstrated by the infertile phenotype in-
duced by neural mutations [27, 39, 40].
Effects of EDC on Sexual Behavior
We here describe the data collected for rodents up to
December 2017, with a special focus on EDC exhibiting
estrogenic/anti-estrogenic or anti-androgenic activities
given the nature of signaling pathways involved in the

Endocrine Disruption of Sexual Behavior Neuroendocrinology 2018;107:400–416 403

DOI: 10.1159/000494558
expression of sexual behavior. Table 1 lists the EDC ana-
lyzed, with their estrogenic or anti-androgenic activities,
uses and reference doses established by agencies when
available. This list, which is far from exhaustive, includes
the most widely studied molecules of these categories.
Furthermore, some molecules, such as bisphenol A (BPA)
or methoxychlor, are listed as estrogenic compounds, but
can actually display anti-androgenic activities or target
the thyroid system as for BPA. The keywords used to
identify the studies concerned included the name of the
molecule of interest (e.g., BPA, phytoestrogen, phthal-
ate), and the terms “sexual behavior,” “endocrine disrup-
tor,” “brain” or “nervous system.” Behavioral data are
presented by period of exposure (developmental periods
including prenatal, postnatal and pre/pubertal develop-
ment, and adult exposure), type of activity of the com-
pound (estrogenic or anti-androgenic), animal species
(rat, mouse), and year of publication, in Tables 2 and 3,
for males and females, respectively. When investigated in
the same studies, neuroanatomical analyses of sexually
dimorphic regions, neuropeptides or receptors involved
in sexual behavior and hormonal measurements are in-
cluded in Tables 2 and 3. Such analyses are otherwise pre-
sented separately in online supplementary Tables 1 and 2
(see www.karger.com/doi/10.1159/000494558 for all on-
line suppl. material) for males and females, respectively.
Effects of Developmental versus Adult Exposure to
Estrogenic Compounds
E2, estradiol benzoate or analogs, such as ethinyl estra-
diol (EE) or diethylstilbestrol (DES), have been used as
positive controls in studies evaluating compounds with
potential estrogenic activity. The behavioral modifica-
tions observed differed between studies, ranging from be-
havior inhibition with blocked ejaculation for estradiol
benzoate in males [41] to unaltered behavior for DES
[42], as shown in Table 2. These discrepancies probably
reflect the different doses of E2 and its analogs used. This,
together with the use of only one dose of the positive con-
trol in most studies, makes it difficult to draw any firm
conclusions about the potential estrogenic effects of the
EDC molecules studied. We will therefore discuss the re-
ported effects of exposure to BPA, phytoestrogens, non-
ylphenol or methoxychlor without comparison to posi-
tive controls.
Effects of Exposure in Males
Prenatal/Postnatal Exposure. Twelve studies in rats
and 7 in mice have investigated the effects of early expo-
sure to estrogenic compounds, such as BPA, phytoestro-
404 Neuroendocrinology 2018;107:400–416
DOI: 10.1159/000494558
gens (coumestrol, genistein, resveratrol) nonylphenol
and methoxychlor (Table 2).
In rats, with the exception of high doses of NP, which
did not affect male sexual behavior [43], changes in at
least one of the components of sexual behavior were re-
ported. Increases in the time to intromission and a larger
number of intromissions were reported for prenatal and
postnatal exposure to BPA, respectively [44]. A dose of
100 µg/kg/day of BPA decreased the intromission ratio
without modifying the number of mounts, mating dura-
tion or time to ejaculation in rats [42], and a decrease in
time to intromission was associated with no change in the
number of intromissions or the intromission ratio [45].
One interesting study reported different behavioral re-
sponses as a function of BPA dose and sexual experience
[46]. Males exposed to phytoestrogens were reported to
have a lower frequency of mounting, intromission and
ejaculation in a number of studies [42, 47–50], whereas 2
studies reported increases in these behaviors [41, 51]. In
middle-aged males, chronic exposure to methoxychlor
has also been shown to decrease time to ejaculation [52].
The published neuroanatomical data are presented in on-
line supplementary Table 1: 6 studies described changes
in the SDN or AVPV volume or cell number [47, 53–57],
whereas 3 studies reported no effect [42, 58, 59]. In the
studies reporting effects of exposure on the organization
of sexually dimorphic populations, both similarities and
differences were observed, depending on the type of neu-
ronal cell analyzed. Indeed, increases in the number of
tyrosine hydroxylase cells or AVPV volume have been
described [47, 56, 57], suggesting demasculinization and/
or feminization of this hypothalamic region. Opposite
patterns have been observed, with either a decrease (res-
veratrol [47]; BPA [54]) or an increase in SDN volume or
calbindin cell number (genistein or nonylphenol [55];
BPA or genistein [57]; BPA [53]). Overall, such neuro-
anatomical measures alone are probably not sufficient or
enough sensitive to reflect EDC-induced effects on the
expression of behavior.
In mice, males exposed prenatally to methoxychlor
displayed diminished sexual arousal, spending less time
close to a female partition [60]. Exposure to BPA or phy-
toestrogen did not affect olfactory preference, time to the
initiation of certain behaviors or the frequency of behav-
ioral events [61–64]. Decatanzaro et al. [61] observed
only a fewer number of intromissions in animals exposed
to BPA in a high-phytoestrogen diet. In addition, male
mice exposed to BPA displayed less lordosis behavior
than the vehicle group when gonadectomized and primed
with E2 and progesterone, indicating a greater masculin-
Mhaouty-Kodja/Naulé/Capela
 Table 2. Behavioral studies in males
Ref. Species Exposure period Route Doses Age at analyses Behavioral analyses Neuroanatomical and hormonal analyses
Developmental
Estrogenic compounds
44 Sprague- Prenatal: from Oral BPA: 40 µg/kg From PDN100 Prenatal exposure: normal preference for females Increased latency to genital sniffing
Dawley rat mating to weaning and intromission
Postnatal: pups from Postnatal exposure: increased number of intromissions
the vehicle group
fostered by BPA-
exposed mothers

42 Wistar rat GD0 to PND21 Oral BPA: 0.1 mg/L (30 µg/kg/day), 11–12 weeks No effect on the number of mounts, mating duration or latency to ejaculation Neither treatment affected the SDN-POA volume
1 mg/L (300 µg/kg/day) Diminution of intromission rate (number intromissions/number mounts ×100) at No effects on LH, FSH or testosterone levels
Trans-RVT: 5 mg/L (1,500 µg/kg/ BPA 0.1 and RVT
day)
DES: 50 µg/L (6.5 µg/kg/day)

Endocrine Disruption of Sexual Behavior

45 Sprague- PND23 to PND30 Oral BPA: 40 µg/kg From PND90 BPA exposure: no effect on the number and intromission ratio. Reduced latency to Reduced testosterone levels at PND37 and PND105 for
Dawley rat EE: 0.4 µg/kg intromission BPA, at PND 37 for EE
EE exposure: reduced duration of genital sniffing, increased number of intromission No effects on estradiol levels
and intromission ratio. Reduced latency to mount and intromit and increased
duration of the refractory period (time between ejaculation and subsequent
mounting)

46 Long- GD7 to PND14 Oral BPA: 5, 50, 500 µg/kg/day or 90–120 days Day 1: increased number of intromissions, reduced time between ejaculation and
Evans rat 5 mg/kg/day subsequent mounting, and reduced latency for the 5 mg group
Day 4: reduced number of intromissions in BPA–5 µg and BPA–50 µg. Increased
latency to intromission and ejaculation at BPA-50 µg, 500 µg and 5 mg. Increased
time between ejaculation and subsequent mounting at 50 µg; copulation more
efficient in controls and BPA–5 mg >BPA–50 µg and 5 mg >BPA–500 µg

65 Deer mice 2 weeks before Oral (diet) BPA: 50 mg/kg diet Since PND80 BPA-exposed males less attractive than controls in mate choice test
mating until
weaning

61 CF-1 mice GD10 to PND9 Oral BPA: 17.5, 175 or 1,750 µg/day in PND 90 No effect on latencies or number of mounts No effects on urinary levels of testosterone and estradiol
high phytoestrogen diet Fewer intromissions in BPA–17.5 and BPA–175 groups
Fewer ejaculation in BPA–17.5 group

63 C57BL/6 GD15 to PND 21 Oral BPA: 50 µg/kg/day, 5 mg/kg/day 8 weeks No effect on latency to mount, thrust, intromit, or ejaculate. Unaffected number of Unaffected number of calbindin neurons in the MPOA
mice (gavage) mounts, intromissions and thrusts and kisspeptin neurons in the AVPV
Reduced lordosis quotient at BPA–5 in gonadectomized males primed with estradiol No effect on testosterone levels
and progesterone

48 Rat PND0 to PND10 or Oral (diet) Coumestrol: 100 µg/g Reduced mount and ejaculation frequency; extended latency to mount and ejaculate No effect on testosterone levels
during PND21 of
lactational period

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49 Sprague- PND0 to PND10 Oral (diet) Coumestrol 100 ppm PND 174–239 Reduced number of mounts (–70%) and ejaculations (–86%) No effect on testosterone levels at PND10, or at 7 or 11
Dawley rat months of age

51 Rat Neonatal Genistein: 20 µg Increased sexual activity

Genistein+benzopyrene 20 µg

50 Long- 2 weeks before Oral (diet) Genistein: 5 mg/kg diet (low) or 70 days Reduced percentage of mounts, intromissions and ejaculations at 5 mg and Reduced testosterone levels at 5 and 300 mg/kg
Evans rat mating until 300 mg/kg diet (high) percentage ejaculation at 300 mg

Neuroendocrinology 2018;107:400–416
weaning

47 Sprague- PND1 to PND22 Oral (water) Resveratrol: 5, 50 or 100 µM PND 132–140 Decreased mount frequency at all doses Reduced SDN-POA volume at all doses; increased AVPV
Dawley rat No effects on frequency, duration or latency of other behaviors (autogrooming and volume at 5 and 50 µM
anogenital investigation) Decreased testosterone levels for all resveratrol doses

41 Wistar rat PND1 to PND5 Subcutaneous Genistein:150 µg Sexual behavior: genistein reduced latency to intromission and ejaculation and
Coumestrol: 5 µg increased frequency of ejaculation; coumestrol induced opposite effects; EB: no
EB: 1 µg ejaculation, increased latency to mount and intromit
Partner preference: no preference of males exposed to coumestrol and genistein in
no-contact test; EB: preference for males; In contact test, genistein-exposed group
preferred receptive females by contrast to coumestrol- and EB-exposed groups

64 C57BL/6 2 weeks before Oral (diet) Genistein: 5 or 300 mg/kg of diet 70 days No effect on latency to mount and intromit, number or duration of mounts and No effect on testosterone levels (tendency to decrease at
mice mating until intromissions the higher dose)
weaning

62 CF-1 mice Gestation Oral (diet) Normal or phytoestrogen-free diet 43 days No effect on mounts or latency to plug

405
 Table 2 (continued)

406
Ref. Species Exposure period Route Doses Age at analyses Behavioral analyses Neuroanatomical and hormonal analyses

101 Swiss mice GD10 to PND40 Oral EE: 0.1 µg/kg/day or 1 mg/kg/day 8 weeks Reduced latency to mount and intromit for EE–1 µg (2nd test) and 0.1 and 1 µg (3rd Increased number of GnRH neurons in the medial
(drinking) (F1, F2, F3 test) for F1–F4 generations; increased frequency of intromission for F2 generation septum at 0.1 µg; reduced GnRH area in the median
and F4) (2nd test) for the two doses and F4 at 0.1 µg and 3rd test for F1–F4 eminence at 0.1 and 1 µg. No effect on kisspeptin neurons
in the hypothalamus at F1, F3 and F4. Increased number
of calbindin neurons in the MPOA of males F1 at 1 µg
(F3, F4) at 0.1 and 1 µg
No effect on testosterone levels for all generations

43 Sprague- PND1 to PND5 Subcutaneous NP: 500 mg/kg PND84 Increased number of mounts for estradiol Unchanged levels of testosterone levels
Dawley rat Estradiol: 2 mg/kg No effect on the number of intromissions in NP or E2 groups

52 Long- PND21 to 11 Oral MXC:200–300–400 mg/kg/day 11 months of The latency to ejaculate was reduced at all doses Serum levels of LH, prolactin and testosterone were not
Evans rat months of age (gavage) age The latency to mount and number of intromissions were not affected affected; heterogeneity among group males were,
however, observed with infertile or sub-fertile males
exhibiting low testosterone levels

60 ICR mice GD5 to GD7 Subcutaneous MXC: 1 mg/0.05 ml (33 mg/kg) 4 months of Males of the MXC-treated group spent less time near the partition after a female in Female introduction in the cage increased plasma
age estrus was introduced into the cage by comparison to untreated or oil-treated groups; testosterone levels in controls, but not in MXC-treated
this suggested altered sexual arousal or motivation in the MXC-treated group groups

Anti-androgenic compounds
93 Long- GD14 to PND3 Oral Vinclozolin (VZ): 1,5, 3, 6 or 12 Reduced erection for all groups; no effect on latency to the first penile reflex;

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Evans rat (gavage) mg/kg increased seminal emission for VZ–12 group

97 Sprague- E8 to E14 IP injection of Vinclozolin: 100 mg/kg/day F3 generation: Control females prefer control males in partner preference test
Dawley rat F0 females 3–4 months

96 Sprague- GD3 to PND21 per os DEHP: 0, 375, 750, or 1,500 mg/ PND 105 Many DEHP exposed males were sexually inactive in the presence of receptive control
Dawley rat kg/day females; sexual inactivity did not correlate with abnormal male reproductive organs

Neuroendocrinology 2018;107:400–416
98 Wistar rat GD6 to PND60 Oral DEHP: 0.015, 0.045, 0.135, 0.405, 110 days: No effects on behavior or fertility Increased testosterone levels at 0.045, 0.405 and 405 mg/
(gavage) 1.215 mg/kg/day or 5, 15, 45, 135, fertility kg/day (2-fold increase in the 405 group)
405 mg/kg/day 130 days:
sexual behavior

94 Sprague- GD1 to PND21 Oral DEHP: 20–100–500 mg/kg/day Adulthood 80% controls, 90% DEHP-20 and -100 and 60% DEHP-500 ejaculated; increased No effect on testosterone levels
Dawley rat (gavage) latency to intromission and intromission number

95 Wistar- GD15 to PND21 Oral (diet) DBP: 20, 200, 2,000 and 20–21weeks Reduced number of mounts and intromissions et 40 ppm DINP and 480 ppm DEHA; No effects on testosterone or estradiol, LH and FSH levels
Imamichi 10,000 ppm no effect of DBP. Reduced number of ejaculations at 200, 2,000 ppm DBP, 40 ppm
rat DINP: 40, 400, 4,000, 20,000 ppm DINP, 48 and 12,000 ppm DEHA; increased number of ejaculations at 10,000 DBP.
DEHA: 480, 2,400, 12,000 ppm Reduced interval post-ejaculation at 10,000 DBP

Adult
Estrogenic compounds
66 Rat Adulthood: 14 days Subcutaneous BPA: 1 mg/rat/day No effect on male sexual behavior Reduced testosterone levels and increased LH levels
of treatment

63 C57BL/6J 8–12 weeks of age Oral BPA: 50 µg/kg/day, 5 mg/kg/day 12 weeks Normal olfactory preference No effect on AR- and ERα-expressing neurons in the
mice (gavage) Increased latency to mount, thrust, intromit and ejaculate in naive and sexually MeA and BNST at 50 µg and 5 mg, but increased number
experienced males at BPA–50 in MPOA at 50 µg and 5 mg
Reduced number of intromissions at BPA–50 No effect on testosterone levels

68 Sprague- Adult: 7-10 days Ground corn- Adult Reduced number of mounts, intromissions, ejaculation, ejaculation latency and
Dawley rat cob bedding grooming frequency

67 Sprague- 7–12 weeks Oral THF-diol isomers from corncob Adulthood Increased latency to mount, intromit and ejaculate nonsignificant
Dawley rat (drinking) bedding: 2 µg/mL Reduced frequency of mount, intromission and grooming

69 Wistar rat 12 weeks of age for Subcutaneous Wet mesquite pod: 3.5 g/kg/day Tests at 10th, Increased latency to mount for Leucaena-, DAI- and E2-exposed groups Reduced testosterone levels at all treatments
50 days Wet plant (Leucaena leaves): 3.5 20th, 30th, Increased latency to intromit except for Leucaena-exposed group
g/kg/day 40th and 50th Increased latency to ejaculate for all groups with strong effect at 40th and 50th days of
Daidzein (DAI): 5 mg/kg/day days of test; increased number of mounts from 20th to 50th day
Genistein: 5 mg/kg/day treatment No effect on number of intromissions; diminished number of ejaculations per test
E2:2 30 µg/kg/day from 20th to 50th day

70 C57BL/6J 8 weeks Oral (diet) NP: 0.5, 5, 50 μg/kg/day 4 weeks later, Normal olfactory preference; increased emission of ultrasonic vocalizations at NP-5; Changes in the number of AR and ERα-immunoreactive
mice exposure increased number of mounts, intromissions and thrusts at NP-5; delayed ejaculation cells in the neural circuitry underlying sexual behavior
maintained No changes in testosterone levels, kisspeptin-
during analyses immunoreactivity in the POA and arcuate nucleus

Mhaouty-Kodja/Naulé/Capela
Anti-androgenic compounds
99 C57BL/6J 8 weeks Oral (diet) DEHP: 0.5, 5, 50 μg/kg/day 4 weeks later, Normal olfactory preference Unaffected colabelling of kisspeptin/AR or ERα in the
mice exposure Reduced emission of ultrasonic vocalizations and male attractiveness; delayed AVPV; reduced number of AR-immunoreactive
maintained initiation of mount and intromission behaviors 09:58 neurones in the POA, BNST and MeA; down-regulation
during Unaffected mating duration or number of mounts and thrusts of AR mRNAs levels in the POA; no effects on ERα
analyses Normal testosterone levels
AVPV, anteroventral periventricular nucleus; BNST, bed nucleus of stria terminalis; EB, estradiol benzoate; GD, gestational day; MeA, medial amygdala; MPOA, medial preoptic area; PND, postnatal day; POA, preoptic area; SDN, sexually dimorphic nucleus.
 Table 3. Behavioral studies in females

Ref. Species Period of exposure Route Doses Age at analyses Behavioral analyses Neuroanatomical and hormonal analyses

Developmental
Estrogenic compounds
73 Sprague- GD11–PND20 Oral (gavage) BPA: 3.2–32–320 mg/kg/day PND10 for the neuroanatomical study No modification of lordosis behavior of OVX+E2/P adult BPA did not modify the volume of the SDN-POA at PND10
Dawley rat DES: 15 μg/kg/day 6 months of age for the behavioral analysis females DES increased SDN volume
(OVX+E2/P).

44 Sprague- Prenatal: Oral (pipette) BPA: 40 μg/kg Adult (PND100) No modification of agonistic behaviors. In proestrus
Dawley rat GD0–PND1 Intact with following of the cycle females, lordosis quotient and proceptive behaviors were
Postnatal: not modified
PND1–PND21 Increased lordosis frequency in BPA-treated proestrus
females

42 Wistar rat GD0–PND21 Oral BPA: 0.1 mg/L Adult (11-12 weeks, OVX+E2/P) BPA: No modification of lordosis quotient or rejection Neither treatment affected the SDN-POA volume
(30 µg/kg/day) – 1 mg/L score in BPA-treated females No effect on LH, FSH or prolactin levels
(300 µg/kg/day) DES and resveratrol: decreased lordosis quotient and Decreased estradiol levels by DES, but not BPA or RVT
DES: 50 µg/L (6.5 µg/kg/day) increased rejection score in DES-treated females
Trans-RVT: 5 mg/L
(1,500 µg/kg/day)

Endocrine Disruption of Sexual Behavior

71 Long- PND0–PND3 Subcutaneous BPA: 50 μg/kg–50 mg/kg Adult (OVX+E2/P) Lordosis quotient abolished in EB-treated females, No effect on hypothalamic GnRH activation
Evans rat PPT: 1 mg/kg unmodified in the other groups
EB: 25 μg

75 Wistar rat PND1–PND7 Subcutaneous BPA: 0.05–20 mg/kg Adult (PND100, No modification of lordosis behavior Reduced ERα expression in the medial preoptic and
OVX+E2/P) Reduced proceptive behaviors in BPA-treated females ventromedial nuclei
No modification of PR expression in these regions

74 Long- GD7–PND18 Oral (gavage) BPA: 2–20–200 μg/kg Adult (OVX+E2/P) Lordosis behavior unchanged following exposure to BPA
Evans rat EE: 15–50 μg/kg/day Abolished behavior by both doses of EE

46 Long- GD7–PND14 Oral (pipette) BPA: 5–50–500 μg/kg–5 mg/kg Adult (intact, behavior during behavioral No modification of sexual receptivity or proceptive
Evans rat estrous) behaviors

72 Long- PND0–PND3 Subcutaneous BPA: 50 μg/kg–50 mg/kg Adult (OVX+E2/P) Proceptive behavior abolished by EB-treated females, Increased number of oxytocin neurons within the
Evans rat PPT: 1 mg/kg unaffected in the other treatment groups paraventricular nucleus in all treatment groups
EB: 25 μg The number of ERα neurons reduced in the medial preoptic
area by EB and PPT, but not by BPA

76 C57BL6/J GD15–PND21 Oral (gavage) BPA: 0.05–5 mg/kg Adult (OVX+E2/P) Increased lordosis quotient in naive females of the BPA– No effect on the number of calbindin-positive cells
mice 0.05 group
No behavioral effects of BPA–5 Increased kisspeptin cell number in the preoptic periventricular
Ovariectomized females supplemented with testosterone nucleus of the rostral periventricular area of the third ventricle
showed comparable number of mounts and thrusts No modification of the kisspeptin fiber density in the arcuate
between vehicle and BPA-groups nucleus. No modification of the number of GnRH neurons in
the medial preoptic area
No modifications in ERα-immunoreactivity in the MeA, BNST
or POA
Increased estradiol levels during diestrus at BPA–0.05 No

DOI: 10.1159/000494558
significant effect on LH levels

51 Rat Neonatal Genistein: 20 mg Genistein significantly increased the lordosis quotient of

females

78 Wistar rats PND1–PND5 Subcutaneous Genistein: 1 mg Adult (OVX+E2) Reduced lordosis quotient in genistein-treated females
Daidzein: 1 mg compared to controls, but higher than that obtained for
17β-E2: 100 μg E2-treated group

Neuroendocrinology 2018;107:400–416
No modification of lordosis quotient in daidzein-treated
females

79 Wistar rat PND5 Subcutaneous Coumestrol: 1–3 mg Adult (OVX+E2) Decreased lordosis quotient of females treated with 3 mg
Genistein: 1 mg coumestrol or E2
E2: 1 mg No modification of lordosis quotient in females treated
with genistein and 1 mg coumestrol

47 Sprague- PND1–PND22 Daily drinking RVT: 5–50–100 μM Adult (OVX+E2/P) No modification of sexual receptivity or proceptive No modification of SDN-POA or AVPV volume
Dawley rat solution behaviors

80 C57BL6/J GD0– Diet Low and high phytoestrogen diet Adult (OVX+E2/P) Decreased lordosis behavior of the aromatase knockout
mice, adulthood females raised on a diet high in phytoestrogens compared
aromatase to both control and aromatase knockout females treated
knockout with a low phytoestrogens diet
mice

77 CF1 mice GD0– Diet Regular laboratory chow or PND43 Strong decrease of lordosis behavior in females fed with a

407
adulthood phytoestrogen-free diet diet deficient in phytoestrogens compared to animals fed
with a regular diet (containing phytoestrogens)
 Table 3 (continued)

408
Ref. Species Period of exposure Route Doses Age at analyses Behavioral analyses Neuroanatomical and hormonal analyses

81 Sprague- PND1, PND3, Subcutaneous MXC: 1 or 2 mg/rat Adult PND75–90 E2 lowered the LQ; both E2 and the higher dose of MXC
Dawley rat PND5 E2: 10 μg/rat (OVX+E2/P) altered the average lordosis amplitude and increased
rejection behavior

Anti-androgenic compounds
95 Wistar- GD15–PND21 Diet DBP: 20–200–2,000–10,000 ppm Adult (day of proestrus) Decreased lordosis behavior in females perinatally No effect on testosterone, estradiol, LH or FSH levels
Imamichi DINP: 40–400–4,000–20,000 ppm exposed to DBP, DINP, DEHA at all the doses
rat DEHA: 480–2,400–12,000 ppm

Adult
Estrogenic compounds
82 Wistar rat Adult OVX Subcutaneous BPA: 10 mg (~40 mg/kg/day) Adult (OVX±P) Ovariectomized (OVX) rats that were primed with 10 mg Like estradiol, injection of BPA at 100 μg, 1,000 μg and 10 mg
(acute) E2: 10 μg (40 μg/kg/day) of BPA, followed by 1 mg of progesterone, displayed doses increased the number of progesterone immunoreactive
rejection behavior, but not lordosis unlike OVX rats cells in the preoptic area and the ventromedial hypothalamic
primed with E2 and progesterone nucleus

89 Long- Adult (5 days Diet Genistein: 13 ppm Adult (OVX+P)±E2 Decreased lordosis quotient in estrogen- and Isoflavone diet increased ERβ mRNA expression in the
Evans rat before testing) Daidzein: 33 ppm progesterone-primed females paraventricular nucleus, by contrast to estradiol which
decreased it

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91 Long- Adult OVX Diet Genistein: 100–500 ppm Adult (OVX+P)±E2 No modification of lordosis quotient by genistein No effect of genistein treatment on oxytocin receptor density in
Evans rat treatment in females primed or not by E2 the ventromedial hypothalamic nucleus
Genistein treatment increased ERβ mRNA expression in the
paraventricular nucleus of the hypothalamus contrary to
17β-estradiol treatment which decreased it

68 Sprague- Adult OVX Ground Adult (OVX+E2/P) Strong decrease of lordosis behavior in females housed
Dawley rat corncob on a corncob bedding

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bedding

86 Sprague- Adult OVX Subcutaneous RVT: 10–100–1,000 μg Adult (OVX+P) RVT treatment did not induce lordosis behavior in Increased E2 levels at resveratrol 1,000 µg and 10 µg EB
Dawley rat (2 days) females

Adult intact: Daily drinking RVT: 100 mM Adult (OVX+E2/P) RVT delayed rejection behavior and lowered the
7 days water frequency of anogenital investigation of stimulus male
preexposure
before OVX

87 Long- Adult OVX Oral Diet: soy isoflavones including Adult (OVX+P 4–5 h before testing) When administered with P alone, diet or tamoxifen had
Evans rat Diet: 5 days (diet±soy) genistein and daidzein no effect
EB: 48 h before Sc (EB, EB: 10 μg/0.1 mL In females primed with both E2 and P, diet or tamoxifen
test tamoxifen) Tamoxifen: 5 mg capsule implant reduced LQ, and hopping and darting rate
Tamoxifen: Soy diet also reduced proceptive behavior
14 days before test

84 Sprague- Adult OVX Oral (gavage) Ferutinin: 0.5 mg/kg Adult (OVX+P)±E2 Ferutinin given alone induced a lordotic response in Ferutinin treatment increased hypothalamic ERα expression
Dawley rat (2, 3, and 4 weeks) ovariectomized rats but failed to affect proceptivity when administered alone, as estradiol did, but decreased the
In combination with estradiol, ferutinin reduced response to estradiol when administered in combination
estrogen-induced receptivity and proceptivity

85 Sprague- Adult OVX Oral (gavage) Ferutinin: 0.2–0.5 mg/kg Adult (OVX+P) Ferutinin given alone at both doses induced preference
Dawley rat (4 weeks) for males, increased lordosis quotient and proceptive
behavior in ovariectomized females like EB

90 Sprague- Adult OVX Oral (gavage) Humulus lupulus extract: Adult (OVX+E2/P) Humulus lupulus extract treatment induced preference
Dawley rat (acute) 5–10–25 mg/kg for males, increased proceptive behaviors at the highest
dose, without affecting lordosis quotient

83 Wistar rat Intact and Subcutaneous Mesquite pod extract (MPE): Adult (intact or OVX) Phytoestrogen and MPE treatment of intact females
OVX adult 4 g/kg decreased the percentage of females exhibiting lordosis
(30 days) Genistein: 1.6 mg/kg by comparison to control and E2 groups
Daidzein: 1.6 mg/kg Genistein and daidzein treatment of OVX females
E2: 40 μg/kg induced lordosis behavior, but lesser than in E2-treated
animals

88 Long- Adult OVX Intramuscular DES: 30–90 μg Adult (OVX+P) Treatment with 30 or 90 μg of DES activated lordosis

Mhaouty-Kodja/Naulé/Capela
Evans rat (27 days) injection (approximately 106–317 μg/kg behavior
respectively)

92 Long- Adult OVX Oral MXC: 200 mg/kg/day Adult (OVX±P) All P-injected rats displayed proceptive (darting) and MXC (400 mg/kg/day) did not affect serum levels of FSH and
Evans rat (24 days) lordosis behaviors by contrast to oil-injected animals LH in intact or OVX females

AVPV, anteroventral periventricular nucleus; EB, estradiol benzoate; MPOA, medial preoptic area; OVX, ovariectomized; P, progesterone; PPT, ERα-selective agonist 4,40,400-(4-propyl-[1H]pyrazole-1,3,5-triyl)trisphenol; SDN, sexually dimorphic nucleus.
ization of behavior [63]. When given a choice between
control and BPA-exposed partners, female mice pre-
ferred controls over BPA-exposed males [65]. BPA had
no neuroanatomical effects on the number of calbindin-
immunoreactive neurons in the preoptic area or kiss-
peptin cells in the AVPV [63].
Thus, exposure to BPA or phytoestrogens induces
changes in the neural circuitry underlying sexual behav-
ior in rats. Little if any change was observed in mice, sug-
gesting species differences, although more studies are re-
quired to confirm these observations. One key observa-
tion is that most of these studies were performed at
relatively low doses, equivalent to or lower than the refer-
ence doses established for these molecules.
Adult Exposure. Exposure to BPA for 2 weeks has no
effect on male rats [66]. By contrast, longer periods of ex-
posure of male mice to BPA increased the times to mount-
ing, intromission and ejaculation, and decreased the
numbers of intromissions and thrusts in both naïve and
sexually experienced male mice [63]. Similar differences
between short- and long-term exposures were observed
for corncob bedding and derived tetrahydrofuran-diols
[67, 68]. Changes in behavior were also reported for rats
subjected to long-term phytoestrogen exposure [69] and
NP [70]. A non-monotonic dose response was observed
for NP since increased emission of ultrasonic vocaliza-
tions, reduced number of mounts, intromissions and
thrust as well as delayed ejaculation were observed only
at the dose of 5 µg/kg/day but not at lower and higher
doses. Despite the small number of studies performed,
the results obtained all suggest that long-term exposure
in adults affects male sexual behavior.
Effects of Exposure in Females
Prenatal/Postnatal Exposure. Female sexual behavior
has mostly been investigated in rats (13 studies vs. only 3
in mice). BPA exposure had no significant effect on lor-
dosis quotient or proceptive behavior in a number of
studies [42, 44, 46, 71–74]. However, pre- or postnatal
exposure to BPA decreased proceptive behavior in one
study [75] and increased the frequency of lordosis in an-
other [44]. In one study on mice, exposure to BPA at a
dose of 50 µg/kg/day resulted in a significant increase in
lordosis quotient in naïve but not sexually experienced
females, whereas a dose 100 times higher had no effect
[76]. Olfactory preferences were unchanged, as BPA-ex-
posed females displayed normal levels of interest in che-
moinvestigating males relative to females [76]. Following
ovariectomy and testosterone treatment, females dis-
played no masculinization of their behavior, with no dif-
Endocrine Disruption of Sexual Behavior
ference in mounting or thrusting behaviors relative to the
vehicle-treated group. Most neuroanatomical studies
have reported no effects on the SDN, corresponding cal-
bindin cells [42, 53, 54, 59, 73, 76], and TH neurons [56].
By contrast, an increase in the number of kisspeptin neu-
rons in the AVPV was observed in the study reporting an
increase in lordosis behavior following BPA exposure in
mice [76]. Changes in the number of tyrosine hydroxylase
neurons have also been reported in rats [54, 56]. These
data strongly suggest that BPA does not trigger masculin-
ization in the female neural circuitry during the highly
sensitive perinatal period, despite its lack of binding to
α-fetoprotein. Instead, it seems to potentiate estrogen-in-
duced increases in sexual behavior and kisspeptin expres-
sion, both of which occur later during the postnatal/pre-
pubertal period in females.
By contrast to BPA, prenatal or postnatal exposure to
phytoestrogens mostly affected adult female behavior, in
both rats and mice, as shown in Table 3 [47, 51, 62, 77–
79]. Similar results have been reported for methoxychlor
[80]. More behavioral and neuroanatomical analyses in
the same context would be required to determine wheth-
er the observed alterations are linked to changes in the
organization of sexually dimorphic populations. Indeed,
no effects of resveratrol on SDN-preoptic area or AVPV
volume were found in a study on female rats in which no
change in behavior was detected [47]. Two other studies
reporting an absence of effect on these two hypothalamic
regions did not assess female behaviors [56, 64].
Adult Exposure. The acute exposure of ovariectomized
rats to BPA did not trigger lordosis behavior when fe-
males were primed with progesterone only, or with pro-
gesterone plus E2, despite an increase in the number of
progesterone-immunoreactive cells in the preoptic area
and ventromedial hypothalamus [81]. An analysis of 10
sets of findings for adult exposure to phytoestrogens sug-
gested that behavioral responses differ between initial
hormonal environments. Hence, genistein and daidzein,
and ferutinin administered to ovariectomized females
primed only with progesterone either induced [82–84] or
had no effect on female behaviors [85, 86]. In similar ex-
perimental conditions, DES treatment induced lordosis
behavior [87], whereas the anti-estrogen tamoxifen had
no effect [86]. By contrast, as for tamoxifen [86], exposure
to genistein and daidzein, corncob bedding, ferutinin,
resveratrol or Humulus lupulus reduced lordosis behav-
ior in 5 studies in which females were either intact or
ovariectomized and on E2 supplementation [68, 82, 83,
86, 88]. In 3 other studies, however, genistein, resveratrol
and H. lupulus had no effect on lordosis behavior but al-
Neuroendocrinology 2018;107:400–416 409
DOI: 10.1159/000494558
Fig. 1. Exposure to endocrine disrupting compounds (EDC) can
alter sexual behavior through indirect (A) and/or direct pathways
(B). A Kisspeptin neurons (green cells) of the preoptic area (POA)
and arcuate nucleus (ARC) activate synthesis and liberation of
GnRH, which stimulates gonadotropin hormone (LH, FSH) lib-
eration, and consequently gonadal synthesis and liberation of sex
steroid hormones. Estradiol and testosterone exert in turn a feed-
back on this gonadotropic axis. Disruption of this axis by EDC may
tered other components of behavior, such as the frequen-
cy of anogenital investigation or proceptive behavior [85,
89, 90]. In the absence of E2, exposure to methoxychlor
also induced proceptive and lordosis behavior in females
after progesterone injection [91]. These data suggest that
phytoestrogens and methoxychlor have estrogenic activ-
ity in the absence of endogenous estrogens, but anti-es-
trogenic effects in the presence of endogenous or supple-
mented estrogens, particularly for phytoestrogens.
Effects of Developmental or Adult Exposure to Anti-
Androgenic Compounds
Five of the 6 studies addressing the developmental ef-
fects of vinclozolin and phthalates at relatively high doses
on components of sexual behavior in males reported be-
havioral changes [92–96]. Diminished erections and in-
creased seminal emission were observed, together with
reduced attractiveness, in rats exposed to vinclozolin [92,
410 Neuroendocrinology 2018;107:400–416
DOI: 10.1159/000494558
interfere with the hormonal dependent regulation of the neural
structures underlying sexual behavior in males and females. B Sim-
plified scheme of the male neural circuitry including the olfactory
bulb (OB), medial amygdala (MeA), bed nucleus of stria termina-
lis (BNST), and preoptic area (POA). EDC can also directly target
these neural structures, to disrupt hormonal signaling pathways
necessary to elicit male sexual behavior.
96]. Exposure to phthalates led to sexual inactivity [95],
increases in the time to sexual behaviors and a decrease
in the frequency of these behaviors in rats [93, 94]. Only
the study by Andrade et al. [97] reported an absence of
effects for low or high doses of DEHP. One neuroana-
tomical study reported no effect on SDN-preoptic area
volume of exposure to high doses of DINP (diisononyl
phthalate) [58]. One recent study addressed the effects of
adult exposure to low doses of DEHP on male sexual be-
havior. Such exposure decreased ultrasonic vocalizations,
reduced female attraction and delayed the initiation of
mating, without affecting olfactory preference for recep-
tive females [98].
Only one study has measured the effects of develop-
mental exposure to phthalates on female sexual behavior.
It reported a decrease in the lordosis behavior of adult
females in proestrus following exposure to DBP (dibutyl
phthalate), DINP, and DEHA (di-[2-ethylhexyl] adipate)
Mhaouty-Kodja/Naulé/Capela
[94]. The effects of adult exposure to anti-androgenic
compounds have not been studied.
More studies of developmental and adult exposure to
anti-androgenic molecules are therefore required, in both
males and females, particularly for doses close to the es-
timated environmental exposure.
Mode of Action
Endocrine disruption of sexual behavior may occur
through indirect or direct pathways (Fig. 1). The indirect
pathway involves changes in the levels of gonadal hor-
mones, which then affect the organization or activation
of neural structures involved in the expression of behav-
ior. The circulating levels of gonadal hormones may be
modified by dysregulation of the hypothalamic GnRH
system and upstream regulators including kisspeptin
neurons and/or the disruption of pituitary function or
gonadal steroidogenesis. Exposure to EDC can also di-
rectly affect the neural structures underlying sexual be-
havior, by interfering with the neural synthesis of hor-
mones, such as the aromatization of testosterone into E2,
binding to and activation of sex steroid receptors, or the
expression of these receptors. Modes of action involving
agonism or antagonism are more difficult to demonstrate
in vivo. They can be suggested or supported by parallel
comparisons with the effects of positive controls, such as
estrogens or their analogs, or anti-androgens, such as flu-
tamide, although such comparisons may be limited for
the reasons described in the section “Effects of Develop-
mental versus Adult Exposure to Estrogenic Com-
pounds”. Analyses of the modes of action of various com-
pounds are reported in Tables 2 and 3 when performed
in parallel to behavioral studies; otherwise, they are
presented separately in online supplementary Tables 1
and 2.
Developmental Exposure
Three of 12 studies on male rats reporting develop-
mental effects of EDC on behaviors described a decrease
in adult hormone levels following exposure to BPA [45],
genistein [50], and resveratrol [47]. The other 9 studies
found no effects on adult levels of testosterone, E2, and
gonadotropins [42, 43, 48, 49, 52, 93, 94]. In mice, no
change in hormone levels was observed following expo-
sure to BPA or EE [61, 63, 99]. Exposure to methoxychlor
did not change basal hormone levels but prevented the
increase in testosterone levels following exposure to fe-
males [60]. The observed impairment of male sexual be-
Endocrine Disruption of Sexual Behavior
havior does not, therefore, seem to be systematically
linked to changes in hormone levels. However, these find-
ings do not rule out the possibility of transient changes in
hormone levels during the exposure period, particularly
if exposure occurs during development, thereby inducing
long-term changes to the neural processes or hormonal
signaling pathways underlying male behavior. Interest-
ingly, long-term changes in the levels of the sex steroid
receptors, ER and PR, have been reported to be induced
by prenatal and postnatal exposure to BPA, DES or me-
thoxychlor in the whole hypothalamus or more specifi-
cally hypothalamic areas (preoptic area, ventromedial hy-
pothalamus), bed nucleus of stria terminalis or medial
amygdala of rats [100–102], and mice [103, 104]. How-
ever, no effects of such exposure were reported in 2 other
mouse studies in the same brain areas [105, 106]. Chang-
es in total hypothalamic gene expression were observed
for BPA and EE, or for neuropeptides involved in social
behavior, such as AVP in the medial amygdala and lat-
eral septum and oxytocin in the hypothalamus [105, 107].
In females, sexual behavior is generally analyzed in
ovariectomized animals supplemented with E2 and pro-
gesterone. The behavioral changes observed under these
experimental conditions cannot be due to modifications
of the hypothalamic-pituitary-gonadal axis. However, it
is important to analyze hormone levels in intact females
in parallel, because they can provide information about
the integrity of the hypothalamic-pituitary-gonadal axis
and its effects on the activation of behavior under physi-
ological conditions. It is not possible to draw any firm
conclusions from the small number of studies assessing
hormone levels in intact females. An absence of effect [42,
105, 108] or increases in hormone levels have been re-
ported following exposure to BPA [76]. Lower levels of
estradiol and LH were observed in rats exposed to me-
thoxychlor [109] or BPA [110], whereas phthalates had
no effect [94]. By contrast, changes in sex steroid receptor
levels have been reported in a larger number of studies.
Most of the studies on BPA presented in online supple-
mentary Table 2 dealt with the effects of exposure on ERα
and/or ERβ levels in neural structures underlying neuro-
endocrine and behavioral reproductive functions such as
the medial amygdala, the bed nucleus of stria terminalis,
the preoptic area and its subdivisions, the ventromedial
hypothalamus or the arcuate nucleus in rats and mice
[100, 101, 103, 105–109, 111–113]. This was probably
motivated by the facts that these receptors are the major
mediators of estrogen-induced genomic regulation, and
effects of EDC exposure on their neural levels can inter-
fere with of the expression of reproduction behaviors.
Neuroendocrinology 2018;107:400–416 411
DOI: 10.1159/000494558
The differences observed in the effects induced may re-
flect differences in the doses of BPA used, the period of
exposure, and the area of the brain analyzed. The few
studies to have assessed ER expression in parallel to be-
haviors reported lower levels of ERα expression in the
preoptic and ventromedial nuclei of females with low lev-
els of proceptive behavior [75], or no change in the num-
ber of ERα-immunoreactive neurons in the medial amyg-
dala, bed nucleus of stria terminalis and preoptic area of
animals with normal or higher levels of proceptive behav-
ior [72, 76]. Exposure to methoxychlor has also been
shown to trigger changes in ER and PR levels in the pre-
optic area [102, 110]. Some studies pointed out the in-
volvement of long-lasting epigenetic modifications in
such changes. Epigenetic gene regulation includes DNA
methylation at CpG sites, post-translational histone
modifications such as methylation or acetylation, and
microRNAs. A previous study showed that changes in-
duced by BPA exposure in the expression levels of ERα in
the hypothalamus are associated with modifications in
DNA methylation in the promoter region of this gene
[103]. Similar observations were made in the preoptic
area of females exposed to methoxychlor [110]. These
findings are of particular interest given the role of ER in
the expression of female sexual behavior. Only a few stud-
ies have monitored the effects of EDC exposure on other
targets, such as neuropeptides essential for the expression
of reproductive behaviors [107, 114].
Adult Exposure
In male rats, changes in male sexual behavior follow-
ing exposure to phytoestrogens were associated with low-
er levels of testosterone [69]. In mice, the impairment of
sexual behavior following chronic adult exposure to BPA,
NP or DEHP was associated with unchanged hormonal
levels and integrity of the hypothalamic-pituitary-gonad-
al axis, together with changes in the level of sex steroid
receptor expression [63, 70, 98]. In particular, DEHP
down-regulated AR protein and mRNA levels in the neu-
ral circuitry involved in sexual behavior [98], while both
the numbers of AR and ERα-immunoreactive cells were
affected by NP exposure [70].
The exposure of adult females to resveratrol or BPA
increased E2 levels in both rats and mice [85, 115]. Inter-
estingly, regardless of its behavioral effects, exposure to
phytoestrogens or BPA increased the levels of ERα, ERβ
or PR expression in the preoptic area, the ventromedial
hypothalamus or the paraventricular nucleus [81, 83, 85,
88, 90, 116].
412 Neuroendocrinology 2018;107:400–416
DOI: 10.1159/000494558
Summary and Conclusions
Several observations can be made based on the data
concerning the impact of EDC on sexual behavior re-
viewed here. In general, sexual behavior appears to be
highly sensitive to EDC, and can be added to the end-
points generally used in assessments of the risks of ex-
posure to EDC and their potential impact on reproduc-
tion. Like other reproductive endpoints, the behavioral
effects induced by exposure to EDC depend on the pe-
riod of exposure. The developmental and pubertal stag-
es are particularly vulnerable due to the organizational
effects of hormones during these periods, but exposure
in adults may also have effects, the doses used in several
studies having elicited effects at doses below the refer-
ence dose. Effects may also depend on the sex of the an-
imals.
The amounts of data published differ considerably be-
tween EDC compounds. The most frequently investigat-
ed compounds are estrogen-like molecules, such as BPA
and phytoestrogens. However, published studies have not
addressed all possible periods of exposure in both sexes.
Developmental exposure to BPA or phytoestrogens has
been extensively studied in both males and females, but
adult exposure to phytoestrogens has been studied only
in females. Much more studies are required for other es-
trogenic or anti-androgenic compounds, for which fewer
data are available, regardless of the exposure period con-
sidered.
Another interesting observation is that changes in the
level of expression of sex steroid receptors (ER, AR, PR)
have been widely documented in both males and females.
This finding strongly suggests that EDC may act, at least
partly, through changes in the neural signaling pathways
underlying sexual behavior, either directly through epi-
genetic modifications to the ERα promoter, as reported
for developmental exposure to BPA [103], or through as
yet unidentified pathways, as for the down-regulation of
AR induced by the exposure of adults to DEHP [98].
Finally, although caution is required when extrapolat-
ing findings from rodents to other species, these altered
processes may be of considerable relevance in both hu-
mans and wildlife. The regulation of libido and erectile
function or reproductive behaviors by sex steroid hor-
mones at the neural level is highly conserved across spe-
cies. In this context, human studies have reported an as-
sociation between a decrease in sexual activity and expo-
sure to environmental doses of DEHP and BPA [117,
118], but the mechanisms underlying these effects remain
unclear.
Mhaouty-Kodja/Naulé/Capela
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