Journal of Atherosclerosis and Thrombosis  Vol.14, No.

5 245

Original Article

Perinatal and Postnatal Exposure to Bisphenol A Increases Adipose

Tissue Mass and Serum Cholesterol Level in Mice

Joji Miyawaki 1 , 3, Kenshi Sakayama 1, Hideo Kato 2, Haruyasu Yamamoto 1, and Hiroshi Masuno 3

1
Department of Bone and Joint Surgery, Ehime University Graduate School of Medicine, Ehime, Japan.
2
Department of Health Sciences, Faculty of Human Culture and Science, Prefectural University of Hiroshima, Hiroshima, Japan.
3
Department of Medical Technology, Faculty of Health Sciences, Ehime Prefectural University of Health Sciences, Ehime, Japan.

Aim: To investigate whether the perinatal and postnatal exposure of mice to bisphenol A (BPA)
caused the development of obesity and/or hyperlipidemia.
Methods: Pregnant mice were exposed to BPA in drinking water at concentrations of either 1 μg/mL
(LD group) or 10 μg/mL (HD group) from gestation day 10 and throughout the lactating period.
After weaning, the pups were allowed free access to drinking water containing the appropriate con-
centrations of BPA. The body weight, adipose tissue weight, and serum lipid levels were measured in
the offspring at postnatal day 31.
Results: In females, the mean body weight increased by 13% in the LD group (p ＜ 0.05) and 11% in
the HD group (p ＜ 0.05) compared with the control group. The mean adipose tissue weight in-
creased by 132% in the LD group (p ＜ 0.01). The mean total cholesterol level increased by 33% in
the LD group (p ＜ 0.01) and 17% in the HD group (p ＜ 0.05). In males, the mean body weight and
mean adipose tissue weight increased by 22% (p ＜ 0.01) and 59% (p ＜ 0.01), respectively, in the HD
group compared with the control group. The mean triacylglycerol level increased by 34% in the LD
group (p ＜ 0.05).
Conclusions: The continuous exposure of mice to BPA during the perinatal and postnatal periods
caused the development of obesity and hyperlipidemia.

J Atheroscler Thromb, 2007; 14:245-252.

Key words; Bisphenol A, Adipose tissue mass, Serum cholesterol, Perinatal exposure

in rodents, there is increasing concern about the nega-

Introduction
Bisphenol A (BPA) is an estrogenic endocrine-
disrupting chemical released by polycarbonate plastics,
such as baby bottles 1), the lacquer coatings of food
cans 2) and dental sealants 3). A small amount of BPA is
found in human serum 4-6). Thus, BPA is present ubiq-
uitously in the environment and ingested routinely by
humans. Since BPA has been associated with reproduc-
tive abnormalities 7), the alteration of patterns of es-
trous cyclicity 8, 9), and the development of embryos 10)
Address for correspondence: Hiroshi Masuno, Department
of Medical Technology, Faculty of Health Sciences, Ehime
Prefectural University of Health Sciences, Takooda, Tobe-cho,
Iyo-gun, Ehime 791-2101, Japan.
E-mail: hmasuno@epu.ac.jp
Received: October 14, 2006
Accepted for publication: April 9, 2007
tive impact of BPA on public health in humans.
Obesity is one of the greatest concerns in public
health, because it is associated with serious morbidi-
ties, including diabetes, hyperlipidemia, hypertension,
cardiovascular disease and osteoarthritis. Obesity is the
result of an increase in body adipose tissue mass pro-
duced by either an enlargement of adipocytes or an in-
creased number of adipocytes or both. We previously
found that BPA not only had the ability to trigger
3T3-L1 fibroblasts to differentiate into adipocytes 11),
but also the ability to accelerate terminal adipocyte
differentiation 12). Sakurai et al. 13) reported that BPA
stimulated glucose transport in 3T3-F442A adipocytes.
These findings suggest that in vivo prolonged expo-
sure to BPA may increase adipose tissue mass and pro-
mote the development of obesity. There are reports
that the perinatal exposure of mice 8) and rats 9) to BPA
246 Miyawaki et al .
caused an increase in body weight. To date, however,
whether this increase in body weight resulted from an
increase in the adipose tissue mass remains unclear.
Seidlová-Wuttke et al.14) reported that the exposure of
ovariectomized rats to BPA did not cause an increase
in paratibial fat storage.
In the present study, we examined whether the
perinatal and postnatal exposure of mice to BPA caused
an increase in body adipose tissue mass, resulting in
the development of obesity. Since obesity is often as-
sociated with hyperlipidemia in humans, the levels of
serum lipids were also measured.
Materials and Methods
Animals and Materials
Pregnant ICR mice were purchased from Clea Ja-
pan Inc. (Tokyo, Japan). A 30% fat diet was purchased
from Oriental Yeast Co., Ltd. (Tokyo, Japan). Triglyc-
eride E-test Wako, cholesterol E-test Wako, NEFA
C-test Wako and glucose CⅡ-test Wako were obtained
from Wako Pure Chemical Industries, Ltd. (Osaka, Ja-
pan). An ELISA kit for mouse/rat leptin was purchased
from B-Bridge International, Inc. (CA, USA).
Experimental Design
Mice were housed in a 12h light- dark cycle (lights
on at 7 am) and allowed free access to food and water.
Since it has been reported that obesity was induced in
mice by exposure to a high-fat diet 15, 16), we used a 30%
fat diet. The diet contained 30% kcal as fat, 55% kcal
as carbohydrate, and 15% kcal as protein with total en-
ergy of 4,353 kcal/kg. The composition of the diet (in
g/kg) was as follows: lard 72.5, soybean oil 72.5, cho-
lesterol 2, sucrose 109, dextrin 150, corn starch 340,
cellulose 40, milk casein 163, vitamin mixture 10, min-
eral mixture 40 and choline chloride 1. We used glass
water bottles to ensure that related compounds did
not leach from plastic water bottles.
Nine pregnant mice were individually housed in
standard polypropylene mouse cages (225×338×140
mm). They were divided into three groups (3 mice/
group) and exposed to BPA in their drinking water at
concentrations of either 1 μg/mL (LD group) or 10
μg/mL (HD group) beginning on day 10 of pregnan-
cy. BPA was dissolved in absolute ethanol and the fi-
nal concentration of ethanol was 0.2%. We gave the
control group drinking water containing only 0.2%
ethanol.
Water intake on days 13 and 16 of gestation was
determined by measuring the difference in the amount
of water placed in the water bottle each day and the
amount remaining the following day, and the levels of
BPA consumed daily were estimated. Daily water in-
take on gestation day 13 and 16 was 11.9±5.8 and
12.6±1.1 mL, respectively, in the control group, 10.5±
3.9 and 13.5±3.7 mL, respectively, in the LD group
and 10.8±5.6 and 16.3±0.9 mL, respectively, in the
HD group. Body weight on gestation day 13 and 16
was 42.5±2.0 and 53.0±2.3 g, respectively, in the
control group, 41.1±0.2 and 51.5±1.1 g, respective-
ly, in the LD group, and 42.9±3.2 and 53.1±4.4 g,
respectively, in the HD group. Based on these values,
we estimated the levels of BPA consumed daily to be
0.26±0.09 (gestation day 13) and 0.26±0.08 mg/kg
body weight (gestation day 16) in the LD group and
2.42±1.12 (gestation day 13) and 3.01±0.89 mg/kg
body weight (gestation day 16) in the HD group. The
average BPA intake was 0.26 mg/kg body weight/day
in the LD group and 2.72 mg/kg body weight/day in
the HD group. The numbers of newborn mice were
14.0±0.6 pups/litter in the control group, 13.3±0.3
pups/litter in the LD group, and 12.7±1.5 pups/litter
in the HD group. BPA exposure continued through-
out the period of lactation. The sexes of the offspring
were initially recorded on postnatal day 8. The num-
bers of females and males were 19 and 23 in the con-
trol group, 16 and 25 in the LD group, and 19 and 19
in the HD group, respectively. The numbers of both
sexes per litter were as follows: 6.3±0.9 females and
7.7±0.9 males in the control group; 5.3±1.5 females
and 8.0±1.5 males in the LD group; 6.3±1.3 females
and 6.3±0.9 males in the HD group. All pups were
transferred with their dam to larger polypropylene
mouse cages (282×451×157 mm) on postnatal day
12. After weaning, the pups were separated from their
dam and allowed free access to a 30% fat diet and wa-
ter containing the appropriate concentrations of BPA.
Thirty days after birth, the food was removed from
the cage at 5 pm, and all mice were killed between 9
and 11:30 am on the following day under ether anes-
thesia. The parametrial adipose tissues of female mice
were removed and weighed. The epididymal adipose
tissues of male mice were removed and weighed.
Animals were treated humanely and with regard
for alleviation of suffering, and the experimental pro-
tocols were reviewed and approved by the local animal
ethics committee at Ehime Prefectural University of
Health Sciences (approval no. 37).
Measurements of Serum Lipids, Glucose and Leptin
The levels of serum total cholesterol (TC), triac-
ylglycerol (TG), non-esterified fatty acid (NEFA) and
glucose were measured using kits for TC, TG, NEFA
and glucose, respectively. Serum leptin was measured
using a leptin ELISA kit.
 Bisphenol A and Adipose Tissue Mass
（A） （B）
25 250
Adipose tissue weight（mg）
20 200
＊
Body weight（g）
＊
15 150
10 100
5 50
0 0
C LD HD
Fig. 1. Effect of BPA on body weight and adipose tissue weight in female offspring.
(A and B) Thirty-one days after birth, overnight fasted female offspring were killed under ether anesthesia,
and their body weight (A) and adipose tissue weight (B) were measured. The values given are the mean ±S.E.
＊
p ＜ 0.05, ＊＊p ＜ 0.01 (compared with the value of the control group). C, control group (n = 19); LD, LD
group (n = 16); HD, HD group (n = 19). (C) Correlation between body weight and adipose tissue weight in
all females (n = 54). (○) control group; (●) LD group; (□) HD group.
Statistical Analyses
Significant differences among three independent
groups were evaluated statistically using one-way anal-
ysis of variance, and subsequent comparisons were
performed using the Tukey-Kramer test, which allows
for unequal sample sizes. Significant differences be-
tween two independent groups were analyzed by Stu-
dent’s t -test. Pearson r was used to calculate correla-
tions between adipose tissue weight and body weight,
and the levels of serum lipids and leptin. All statistical
procedures were carried out using SPSS software (ver-
sion 13.0). A p value of ＜ 0.05 was considered to in-
dicate significant difference. All values were expressed
as the mean±SE.
Results
Effect of BPA on Body Weight and Adipose Tissue
Mass
Pregnant mice were exposed to BPA from gesta-
tion day 10 and throughout the lactating period. After
weaning, the pups were separated from their dams and
exposed to BPA. Thirty-one days after birth, their body
weights and adipose tissue weights were measured.
In females, the mean body weight increased by 13%
in the LD group and 11% in the HD group compar-
ed with the control group [F(2,51) = 5.306, p ＜ 0.01]
(Fig. 1A). The mean adipose tissue weight of the LD
group increased by 132% compared with the control
group, while the weight of the HD group did not dif-
fer significantly from the control group [F(2,51) =
9.883, p ＜ 0.01] (Fig. 1B). The percentage of adipose
tissue weight to body weight was significantly higher
in the LD group, but not the HD group, than in the
247
（C）
300
Adipose tissue weight（mg）
250
＊＊ 200
150
100
50 r ＝0.810
p＜0.0001
0
C LD HD 10 12 14 16 18 20 22 24
Body weight（g）
control group [0.36±0.03% in the control group;
0.71±0.09% in the LD group (p ＜ 0.01, vs the con-
trol group); 0.50±0.04% in the HD group; F(2,51) =
10.811, p ＜ 0.001]. When the correlation between ad-
ipose tissue weight and body weight was examined in
all females, adipose tissue weight showed a strong pos-
itive correlation with body weight (Fig. 1C). This re-
sult indicates that increased body weight resulted from
an increase in adipose tissue mass.
In males, the mean body weight and mean adi-
pose tissue weight of the HD group increased by 22%
and 59%, respectively, compared with the control
group, while these two weights in the LD group did
not differ significantly from the control group [body
weight, F(2,64) = 12.069, p ＜ 0.001; adipose tissue
weight, F(2,64) = 10.904, p ＜0.001] (Figs. 2A and 2B).
The percentage of adipose tissue weight to body weight
was significantly higher in the HD group, but not
the LD group, than in the control group [0.61±0.03%
in the control group; 0.66±0.03% in the LD group;
0.79±0.05% in the HD group (p ＜ 0.01, vs the con-
trol group); F(2,64) = 6.505, p ＜ 0.01]. As in females,
the adipose tissue weight of males also correlated posi-
tively with body weight (Fig. 2C).
Effect of BPA on Serum Leptin Levels
Since leptin is a product of the ob gene and a hor-
mone secreted by adipocytes with an important func-
tion in the regulation of the amount of body fat 17), the
level of serum leptin was measured. In females, the
mean leptin level of the LD group increased by 123%
compared with the control group, while the level of
the HD group did not differ significantly from the
control group [F(2,51) = 3.449, p ＜0.05] (Fig. 3A). The
248 Miyawaki et al .

（A） （B） （C）

25 250 300

Adipose tissue weight（mg）

Adipose tissue weight（mg）

20 ＊＊ 200 250
Body weight（g）

＊＊
200
15 150
150
10 100
100
5 50 50 r ＝0.842
p＜0.0001
0 0 0
C LD HD C LD HD 10 12 14 16 18 20 22 24
Body weight（g）

Fig. 2. Effect of BPA on body weight and adipose tissue weight in male offspring.
(A and B) Thirty-one days after birth, overnight fasted male offspring were killed under ether anesthesia,
and their body weight (A) and adipose tissue weight (B) were measured. The values given are the mean ±S.E.
＊＊
p ＜ 0.01 (compared with the value of the control group). C, control group (n = 23); LD, LD group
(n = 25); HD, HD group (n = 19). (C) Correlation between body weight and adipose tissue weight in all
males (n = 67). (○) control group; (●) LD group; (□) HD group.

leptin levels of females showed a positive correlation （A）Female （B）Female

with adipose tissue weight (Fig. 3B). 5 10
In males, no differences in mean leptin levels were ＊
Serum leptin（ng/mL）

Serum leptin（ng/mL）
observed among the three groups (Fig. 3C), and leptin 4 8

levels did not show any significant correlation with ad- 3 6

ipose tissue weight (Fig. 3D).
2 4

Effect of BPA on Serum Lipid and Glucose Levels 1 2

r ＝0.603
Table 1 shows the levels of serum TC, TG, NEFA p＜0.0001
0 0
and glucose. In females, the mean TC level increased C LD HD 0 50 100 150 200 250 300
by 33% in the LD group and 17% in the HD group Adipose tissue weight（mg）
compared with the control group [F(2,51) = 8.645, p ＜
0.001]. There were no differences in the mean TG,
NEFA and glucose levels among the three groups. （C）Male （D）Male
In males, the mean TC level in LD and HD 5 10
groups tended to be higher, but not significantly, com-
Serum leptin（ng/mL）

Serum leptin（ng/mL）

pared with the control group. The mean TG and
NEFA levels of the LD group increased by 34 and
29%, respectively, compared with the control group,
while the levels of the HD group did not differ signif-
icantly from the control group [TG, F(2,64) = 9.674,
p ＜ 0.001; NEFA, F(2,64) = 18.171, p ＜ 0.001]. The
mean glucose level of the LD group, but not the HD
group, decreased by 41% compared with the control
group [F(2,64) = 4.667, p ＜ 0.05].
Correlation between Adipose Tissue Weight and the
Levels of Serum Lipids and Glucose
Table 2 shows the correlation coefficients be-
tween adipose tissue weight and the levels of serum
lipids and glucose. In females, the levels of serum TC,
TG and glucose correlated positively with adipose tis-
sue weight, while the level of serum NEFA correlated
negatively. As in females, the levels of serum TC and
4 8 r ＝0.180
N.S.
3 6
2 4
1 2
0 0
C LD HD 0 50 100 150 200 250 300
Adipose tissue weight（mg）
Fig. 3. Effect of BPA on the level of serum leptin.
(A and C) Levels of serum leptin in offspring were measured using
an ELISA kit for leptin. The values given are the mean±S.E. Panel
(A) shows the serum leptin level of female offspring. ＊p ＜ 0.05
(compared with the value of the control group). C, control group
(n = 19); LD, LD group (n = 16); HD, HD group (n = 19). Panel (C)
shows the serum leptin level of male offspring. C, control group
(n = 23); LD, LD group (n = 25); HD, HD group (n = 19). (B and
D) Correlations between the leptin level and adipose tissue weight
in female offspring (B) and male offspring (D) are shown. (○) con-
trol group; (●) LD group; (□) HD group. N.S., not significant.
 Bisphenol A and Adipose Tissue Mass 249

Table 1. Serum TC, TG, NEFA and glucose levels in female and male mice
Female mice Male mice
Control group LD group HD group Control group LD group HD group
(n = 19) a (n = 16) (n = 19) (n = 23) (n = 25) (n = 19)
Serum TC (mg/dL) 165±8 b 219±11＊＊ 193±8＊ 163±5 201±10 193±12
Serum TG (mg/dL) 87±5 98±12 79±4 79±6 106±8＊ 66±4
Serum NEFA (mEq/L) 2.23±0.05 2.36±0.18 2.12±0.08 2.02±0.07 2.61±0.11＊＊ 1.91±0.07
Serum glucose (mg/dL) 86±12 80±12 66±6 84±8 50±10＊ 83±9
TC, total cholesterol; TG, triacylglycerol; NEFA, non-esterified fatty acid
a
Numbers of parentheses show the number of mice.
b mean±SE
＊
p ＜ 0.05, ＊＊p ＜ 0.01 (compared with the value of the control group)

Table 2. Correlation coefficients between adipose tissue weight
and the levels of serum lipids and glucose
Adipose tissue weight (mg)
female p-value male p-value
Serum TC (mg/dL) 0.570 ＜0.0001 0.283 0.0212
Serum TG (mg/dL) 0.301 0.0270 −0.377 0.0018
Serum NEFA (mEq/L) −0.312 0.0231 −0.353 0.0039
Serum glucose (mg/dL) 0.311 0.0219 0.399 0.0009
TC, total cholesterol; TG, triacylglycerol; NEFA, non-esterified fatty
acid
glucose of males showed a positive correlation with
adipose tissue and the level of serum NEFA showed a
negative correlation. Surprisingly, the level of serum
TG in males showed a negative correlation with adi-
pose tissue weight.
Discussion
The aim of this study was to determine whether
BPA is involved in the development of obesity and/or
hyperlipidemia. In this study, we gave pregnant mice
drinking water containing either 1 μg/mL or 10 μg/
mL of BPA from gestation day 10 and throughout the
lactating period, and then gave the pups drinking wa-
ter containing appropriate concentrations of BPA after
weaning. The measurement of body weight at postna-
tal day 31 showed that continuous exposure to BPA
caused an increase in body weight in both female and
male offspring. This agrees with the finding of Rubin
et al.9) that both female and male offspring born to
rats exposed to BPA exhibited an increase in body
weight at postnatal day 28. They gave pregnant rats
drinking water containing the same concentrations of
BPA as those used in this study from gestation day 6
to postnatal day 21, and then gave the pups unadul-
terated drinking water after weaning.
Since obesity is characterized by the accumulation
of excessive adipose tissue mass in the body, we mea-
sured the parametrial/epididymal adipose tissue weight.
Our data on adipose tissue weight demonstrated that
BPA caused a nonmonotonic and inverted-U-shaped
dose-response increase in the mean adipose tissue mass
in female offspring, while this chemical caused a mono-
tonic dose-response increase in the mass in male off-
spring. The ratios of adipose tissue mass to body weight
were also increased in similar dose-response manners.
Moreover, simple regression analysis showed that, in
both female and male offspring, body weight increased
with the increase in adipose tissue mass. These results
suggest the involvement of BPA in the development of
obesity in both female and male offspring. This inter-
pretation raises the question of whether BPA affects
other parameters of obesity.
Most cases of obesity in rodents 16, 18, 19) and hu-
mans 20-22) are associated with high circulating leptin
levels. For example, the serum leptin levels of obese
children correlated highly with arm fat and body mass
index 21); therefore, to explore the above-mentioned
question, we measured the serum leptin levels. In fe-
males, serum leptin levels were significantly higher in
pups born to mice exposed to BPA than in those born
to control mice. As for adipose tissue mass, mean
leptin levels increased in a nonmonotonic and invert-
ed-U-shaped dose-response manner, resulting in a pos-
itive correlation between serum leptin levels and adi-
pose tissue mass. Based on these findings, we conclud-
ed that the continuous exposure of mice to BPA dur-
ing the perinatal and postnatal periods caused the de-
velopment of obesity in female offspring. To our knowl-
edge, this is the first report demonstrating the increas-
ing effect of BPA on adipose tissue mass and serum
leptin level. Takeuchi et al.23) reported that serum BPA
concentrations were higher in young obese women
250 Miyawaki et al .
than in young non-obese women and correlated posi-
tively with body mass index. This finding suggests that
BPA might be associated with the development of
obesity in young women.
In males, BPA failed to increase serum leptin lev-
els, resulting in no correlation between the serum
leptin level and adipose tissue mass. Two possibilities
for this failure were considered. One possibility is the
lower increasing effect of BPA on adipose tissue mass
in males than in females. For example, BPA caused a
132% increase in mean adipose tissue weight in the
LD group of females, while it caused only a 59% in-
crease in weight in the HD group of males. Another
possibility is the gender differences in leptin produc-
tion in adipose tissue. We observed in the control
group that adipose tissue was heavier in males than in
females [males (n = 23), 95.2±5.6 mg; females (n = 19),
56.1±4.5 mg; p ＜ 0.01], whereas there were no differ-
ences in serum leptin levels between genders [males
(n = 23), 1.36±0.21 ng/mL; females (n = 19), 1.51±
0.26 ng/mL]. Frederich et al.24) also reported that when
fed the high-fat diet, equivalent levels of plasma leptin
were observed in female and male mice despite female
mice having less total body lipid. They described that
if female mice produced more leptin at any fat cell
size, and the hypothalamus responded to leptin in an
identical manner in both sexes, reduced body fat con-
tent in the presence of equivalent leptin levels would
result. Ahrén 25) reported that the plasma leptin level
displayed a nocturnal increase in mice and that this
increase was 2.6-times greater in females than in males.
Thus, the ability to produce leptin in adipose tissue
was lower in males than in females.
In humans, obesity often develops secondarily to
hyperlipidemia and cardiovascular disease. Cardiovas-
cular disease is the major cause of morbidity and mor-
tality, and the risk of coronary heart disease is associat-
ed with high circulating TC levels 26-28). To our knowl-
edge, there are only two reports on the effect of BPA
on serum lipid levels. Dodge et al. 29) reported that the
4-day oral administration of BPA (0.1, 1.0 or 10 mg/
kg body weight/day) to ovariectomized rats (6 months
old) did not change the serum TC level, but BPA at
the highest dose (30 mg/kg body weight/day) decreased
it. Seidlová-Wuttke et al.14) reported that the 3-month
administration of pellet food containing BPA (0.033
or 0.333 mg/kg body weight/day) to ovariectomized
rats (3 months old) did not change serum TC and TG
levels. We recently observed that the 30-day adminis-
tration of a 30% fat diet containing BPA (approxi-
mately 0.19 or 18.9 mg/kg body weight/day) to male
ICR mice (4 weeks old) did not cause any changes in
serum TC and TG levels (H Masuno - unpublished
data). Thus, the administration of BPA at doses of
＜ 20 mg/kg body weight/day during adulthood did
not affect serum lipid levels. In the present study, we
administered BPA (0.26 or 2.72 mg/kg body weight/
day) to mice during the perinatal and postnatal peri-
ods and measured serum lipid levels on postnatal day
31. In females, BPA increased the serum TC level in a
nonmonotonic and inverted-U-shaped dose-response
manner, but did not cause a significant increase in the
serum TG level. The serum TC level showed a positive
correlation with adipose tissue weight. This is compat-
ible with the observation that obesity is often accom-
panied with hyperlipidemia. In males, BPA increased
the serum TG level in a nonmonotonic and inverted-
U-shaped dose-response manner, but did not cause a
significant increase in the serum TC level. Surprising-
ly, the serum TG level correlated negatively with the
adipose tissue weight. This is inconsistent with the
above description. Since the serum TG level correlated
positively with the serum NEFA (r = 0.913, p ＜ 0.0001)
and negatively with the serum glucose level (r = −0.541,
p ＜ 0.0001), this suggests that the high level of serum
TG might be due to the overproduction of TG from
NEFA, but not glucose. Based on these findings, we
concluded that the continuous exposure of mice to
BPA during the perinatal and postnatal periods caused
the development of hypercholesterolemia in female off-
spring and of hypertriglyceridemia in male offspring.
Although the mechanism by which perinatal and
postnatal exposure to BPA elicits increasing effects on
the adipose tissue mass and serum lipids differently in
female and male offspring is unclear, there have been
some reports on the differences in responses to BPA
between female and male offspring born to exposed
dams. Rubin et al.9) reported that the increase in body
weight of rats born to dams exposed to BPA persisted
longer in female offspring than in male offspring after
BPA administration ceased. Farabollini et al.30) de-
scribed that perinatal exposure of rats to BPA in the
period of sexual differentiation of the brain influenced
adult behavior differently in females and males.
Our present data showed that low-dose BPA was
more effective than high-dose BPA in altering some
parameters of obesity and hyperlipidemia. This is con-
sistent with the finding of Rubin et al.9) that, in female
rats exposed perinatally to BPA, the lower of the two
BPA doses (0.1 and 1.2 mg/kg body weight/day) pro-
duced a larger effect on body weight relative to the
higher dose. Similar nonmonotonic and inverted-U-
shaped dose-response curves of BPA were observed in
measurements of preputial gland weight 31) and testis
weight 32) in rodents. vom Saal et al.31) described that
responses to weak environmental estrogens, including
 Bisphenol A and Adipose Tissue Mass
BPA, cannot be assumed to be monotonic across a
wide dose range. Moreover, they found that estradiol
and diethylstilbestrol also increased prostate weight in
an inverted-U-shaped dose-response manner in mice 33).
BPA is used in the food-packaging industry and
in dentistry, and leaches from the lining of tin cans
into foods and from dental sealants into saliva. For
example, microgram amounts of BPA (4-23 μg/can)
were found in both extracted foods and water from
autoclaved cans 2). Olea et al.3) detected 90-931 μg of
BPA in the saliva during the first 1-h after the applica-
tion of dental sealant (50 mg). The maximum amount
(931 μg) was reported to represent 0.0133 mg/kg body
weight for a person weighing 70 kg and 0.0372 mg/kg
body weight for a child weighing 25 kg 29). The low-
dose BPA used in the present experiment was close to
that used by Rubin et al.9), but was about 10-times
higher than a dose equivalent to that found in saliva.
Howdeshell et al.8) reported that the perinatal expo-
sure of mice to very low-dose BPA (0.0024 mg/kg
body weight/day) increased body weight on the wean-
ing day in female offspring. This finding suggests that
BPA at lower doses than those used in the present
study might increase body adipose tissue mass.
In conclusion, the perinatal and postnatal expo-
sure of mice to BPA increased body adipose tissue mass
and serum TC and TG levels. The increasing effect of
BPA on adipose tissue mass was more pronounced in
females than in males.
Acknowledgements
This work was supported in part by a Grant-in-
Aid for Scientific Research (C) from the Japan Society
for the Promotion of Science (H.M.) and a grant
(Kyouiku Kenkyu Joseihi) from Ehime Prefectural
University of Health Sciences.
We are grateful to Dr. Hiroyuki Yoshimura (Be-
havioral Pharmacology Laboratory, Faculty of Health
Sciences, Ehime University School of Medicine, Ehime)
for advice on statistical analyses.
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