Am J Physiol Endocrinol Metab 303: E1061E1068, 2012.

First published August 28, 2012; doi:10.1152/ajpendo.00213.2012.

Perinatal exercise improves glucose homeostasis in adult offspring

Lindsay G. Carter,1 Kaitlyn N. Lewis,2,3 Donald C. Wilkerson,1,4 Christine M. Tobia,1
Sara Y. Ngo Tenlep,1 Preetha Shridas,5 Mary L. Garcia-Cazarin,6 Gretchen Wolff,6
Francisco H. Andrade,6 Richard J. Charnigo,7 Karyn A. Esser,6 Josephine M. Egan,8 Rafael de Cabo,2
and Kevin J. Pearson1
1

Graduate Center for Nutritional Sciences, College of Medicine, University of Kentucky, Lexington, Kentucky; 2Laboratory of
Experimental Gerontology, National Institute on Aging, National Institutes of Health, Baltimore, Maryland; 3Barshop Institute
for Longevity and Aging Studies, Departments of Cellular and Structural Biology and Physiology, University of Texas Health
Science Center at San Antonio, San Antonio, Texas; 4Owensboro Cancer Research Program, Owensboro, Kentucky;
5
Department of Internal Medicine, College of Medicine, University of Kentucky, Lexington, Kentucky; 6Department of
Physiology, College of Medicine, University of Kentucky, Lexington, Kentucky; 7Department of Biostatistics, College of
Public Health, University of Kentucky, Lexington, Kentucky; and 8Laboratory of Clinical Investigation, National Institute on
Aging, National Institutes of Health, Baltimore, Maryland
Submitted 26 April 2012; accepted in final form 26 August 2012

Carter LG, Lewis KN, Wilkerson DC, Tobia CM, Tenlep SY,
Shridas P, Garcia-Cazarin ML, Wolff G, Andrade FH, Charnigo
RJ, Esser KA, Egan JM, de Cabo R, Pearson KJ. Perinatal exercise
improves glucose homeostasis in adult offspring. Am J Physiol Endocrinol Metab 303: E1061E1068, 2012. First published August 28,
2012; doi:10.1152/ajpendo.00213.2012.Emerging research has shown
that subtle factors during pregnancy and gestation can influence
long-term health in offspring. In an attempt to be proactive, we set out
to explore whether a nonpharmacological intervention, perinatal exercise, might improve offspring health. Female mice were separated
into sedentary or exercise cohorts, with the exercise cohort having
voluntary access to a running wheel prior to mating and during
pregnancy and nursing. Offspring were weaned, and analyses were
performed on the mature offspring that did not have access to running
wheels during any portion of their lives. Perinatal exercise caused
improved glucose disposal following an oral glucose challenge in both
female and male adult offspring (P 0.05 for both). Blood glucose
concentrations were reduced to lower values in response to an intraperitoneal insulin tolerance test for both female and male adult
offspring of parents with access to running wheels (P 0.05 and P
0.01, respectively). Male offspring from exercised dams showed
increased percent lean mass and decreased fat mass percent compared
with male offspring from sedentary dams (P 0.01 for both), but
these parameters were unchanged in female offspring. These data
suggest that short-term maternal voluntary exercise prior to and
during healthy pregnancy and nursing can enhance long-term glucose
homeostasis in offspring.
running; pregnancy; programming; mice

in the US were estimated to have

diabetes, and this number is increasing (4). Interestingly, and
what is often underappreciated, is that the metabolic status of
an individual is decided not only by their inherited genes,
nutritional intake, and physical exercise but also by maternal
nutrition and obesity during pregnancy. In 1992, Hales and
Barker (18) put forth the thrifty phenotype hypothesis that
suggested that malnourished pregnant mothers produce smaller
offspring that have a higher incidence of obesity, diabetes, and
heart disease in adulthood. This hypothesis has since been

IN 2007, 23.5 MILLION PEOPLE

Address for reprint requests and other correspondence: K. J. Pearson,

Graduate Center for Nutritional Sciences, Univ. of Kentucky, Wethington
Bldg., Rm. 591, 900 South Limestone St. Lexington, KY 40536-0200 (e-mail:
kevin.pearson@uky.edu).
http://www.ajpendo.org

modified to the developmental origins of health and disease

(DOHaD) (15, 17, 18).
The DOHaD suggests that the maternal environment and
fetal programming lead to a higher incidence of several diseases later in life (14, 16). A growing number of studies have
been designed to provide evidence for the negative impact of
DOHaD, using mice, rats, and sheep as animal models (11, 12,
31, 38). Many of these studies are directed at malnutrition
through protein restriction or physical stressors that produce
similar effects (8, 11, 34, 44, 45), but more recent studies are
elucidating the metabolic effects of high-fat diet consumption
during pregnancy on offspring (22, 38, 44, 48).
It has been known since Hippocrates and Galen that physical
activity is an important component of a healthy lifestyle (33).
However, knowledge about the contributions of maternal exercise during pregnancy and the long-term consequences on
offspring is minimal. In both rats and mice, maternal exercise
during pregnancy can improve brain physiology and cognition
in the offspring (2, 23, 25). In humans, 5-yr-old children born
to mothers who exercised regularly during pregnancy had
improved intelligence scores and reduced body mass (7).
Physical activity is already used as a treatment for gestational
diabetes in humans, but long-term outcomes in offspring have
not been fully investigated (10, 28, 41, 49).
Using a mouse model, we set out to explore maternal
voluntary exercise as an intervention to improve offspring
metabolic health. We hypothesized that voluntary exercise
prior to and during pregnancy and nursing would benefit
offspring metabolic health throughout their adult life. In this
report, we show that maternal voluntary exercise just prior to and
during pregnancy and lactation had a positive impact on glucose
regulation and insulin sensitivity in sedentary adult offspring. This
is exciting because it indicates that a simple, short-term, nonpharmacological intervention can improve long-term glucose homeostasis in the next generation.
RESEARCH DESIGN AND METHODS

Animals and diets. These studies were carried out at the University
of Kentucky according to an approved Institutional Animal Care and
Use Committee protocol. At 2 mo of age, female Institute for Cancer
Research (ICR) mice were bred and produced one litter at Taconic
prior to shipment to the University of Kentucky at 4 mo of age.
Females were housed four mice per cage for a 2-wk acclimation
E1061

E1062

PERINATAL EXERCISE IMPROVES OFFSPRING HEALTH

period and then separated into two groups, sedentary or voluntary

exercise, and were housed singly for the duration of the study. They
were placed in light-controlled boxes (Phenome Technologies, Lincolnshire, IL) in an environmentally controlled vivarium between 68
and 72F, with unlimited access to food and water under a controlled
photoperiod (14:10-h light-dark). The dams (and sires) were fed
Labdiet Formulab Diet No. 5008. Maternal body weight and food
intake were measured once/wk throughout the breeding portion of the
study. The pregnant dams in the exercise cohort had continual access
to running wheels throughout pregnancy until 14 days after giving
birth, at which point wheels were removed to prevent possible harm
to growing pups. Litters were culled to eight or nine pups 48 h after
birth. Pups were cross-fostered from other litters from the same group
if they did not have at least eight pups. Pups were weaned on postnatal
day 21 onto Teklad Global 18% Protein Rodent Diet No. 2018 and
were housed four to five mice per cage. The offspring themselves did
not have access to running wheels during any portion of the study.
Subsequent analyses were performed on the offspring from 20 and 18
different sedentary and exercised nursing dams, respectively.
Exercise. Female ICR mice were housed in cages purchased from
Phenome Technologies (Lincolnshire, IL). The mice had open access
to the running wheels that were mounted within each cage. A
mechanical counter was used to record wheel rotations to a desktop
computer via ClockLab software (Actimetrics, Wilmette, IL). Sedentary female mice were housed in nearly identical cages that did not
contain running wheels. Dams exercised 7 days prior to and during
pregnancy and up through day 14 of lactation. Male mice also had the
ability to exercise for the 10-day period while they were in the female
cages for mating purposes. The exercise was completely voluntary;
mice were not forced in any way onto wheels. Offspring did not
exercise for any portion of the study.
Oral glucose tolerance test. At 3132 (6-h fast), 36 37 (3-h fast),
and 7172 (3-h fast) wk of age, offspring of both sexes were fasted
and then given an oral gavage of D-()-glucose (Sigma-Aldrich, St.
Louis, MO) at 2 g/kg body wt. Blood glucose was measured in tail
vein blood using an Ascensia Breeze 2 meter (Bayer, Mishawaka, IN)
just prior to gavage (time 0) and at 15, 30, 60, and 120 min after
glucose administration.
Intraperitoneal insulin tolerance test. At 3334 (female) and
43 44 (male) wk of age, mice were fasted for 3 h. Female mice were
then given an intraperitoneal injection of porcine insulin (Sigma,
I-5523) at 0.75 IU/kg body wt. Male mice were given an intraperitoneal insulin injection at 1.25 IU/kg body wt. Blood glucose was
measured at 0, 15, 30, 60, 120, and 180 min postinjection from tail
vein prick. Female mice that became unresponsive to touch as a result
of hypoglycemia were given a glucose injection (n 2/20 for
sedentary and n 5/18 for exercise). No further blood glucose
readings were taken from the glucose-injected mice, but preglucose
injection values were included in the analysis. No male mice became
unresponsive during the procedure.
In vitro soleus muscle glucose uptake. Female offspring born to
sedentary and exercise dams were fasted for 3 h at 37 wk of age (n
6/group). Mouse soleus muscles were then quickly excised immediately after euthanasia. The tissue was immersed in Krebs-Ringer
bicarbonate buffer (117 mM NaCl, 4.7 mM KCl, 24.6 mM NaHCO3,
1.2 mM KH2PO4, 1.2 mM CaCl2, and 2.5 mM MgSO4) bubbled with
95% O2 and 5% CO2. Soleus muscles from one leg were used to
measure basal glucose uptake, and the contralateral soleus muscles
were used to measure the effects of insulin. All muscles were first
incubated with Krebs-Ringer bicarbonate buffer with 2 mM pyruvate
for 30 min at 37C. Soleus muscles were then rinsed and incubated
with Krebs-Ringer buffer containing 1 mM 2-deoxy-D-[1,2-3H]glucose ([2D-3H]glucose, 1.5 mCi/ml) and 7 mM D-[14C]mannitol (0.45
mCi/ml) for 10 min. Insulin (100 nM) was added to the buffer of the
insulin group. Finally, soleus muscles were rinsed with plain KrebsRinger buffer. Tendons were removed and soleus muscles blotted dry
with filter paper and digested with 250 l of 1 N NaOH at 80C for

10 min. After neutralizing with 250 l of 1 N HCl, 350 l of sample

was added to scintillation liquid for dual label radioactivity counting.
Glucose uptake (per gram tissue weight) was then determined after the
intracellular and extracellular space was calculated, as described
previously by Chambers et al. (5). Tissues from male offspring were
not tested due to time constraints.
In vitro adipose glucose uptake. A separate cohort of female
offspring born to sedentary and exercise dams was fasted for 3 h at 36
wk of age (n 4/group). Glucose uptake was performed as described
previously with some modifications (37). Briefly, parametrial fat was
removed immediately after euthanasia and cut into 5- to 10-mg sized
explants. Explants were washed three times in 0.5 ml of Krebs-Ringer
buffer (pH 7.4) supplemented with 1% BSA. Explants were incubated
in Krebs-Ringer buffer with 1% BSA at 37C for 30 min to establish
basal conditions. Explants were then transferred to a 24-well plate
containing 450 l of Krebs-Ringer buffer with either 0 or 100 nM
insulin and incubated for 15 min at 37C under 5% CO2. The assay
was initiated by the addition of 50 l of 4.5 mM 2-deoxyglucose
containing 0.5 Ci of 2-[14C]deoxyglucose (57.7 mCi/mmol,
NEC495A00; Perkin-Elmer). After 15 min, the assay was terminated
by transferring tissue explants to ice-cold Krebs-Ringer buffer supplemented with 1% BSA, washed three times with the same buffer,
blotted, weighed, and incubated in 1 N NaOH for 1 h at 65C.
Radioactivity of the NaOH extract was determined by scintillation
counting. Glucose uptake is expressed per gram tissue weight. Tissues
from male offspring were not tested due to time constraints.
Body composition. At 39 and 68 wk of age, total fat tissue, lean
tissue, and water were measured in live, conscious male and female
offspring by nuclear magnetic resonance (EchoMRI; EchoMedical
Systems, Houston, TX). The EchoMRI measures adipose tissue, lean
mass, and free and total water. Although many tissues contribute to
the lean mass output, there are undetectable components such as bone
mineral content, hair, and claws.
Statistics. Repeated-measures data were analyzed using repeatedmeasures analyses of variance (ANOVA), followed by Students
t-test, unless otherwise indicated. Repeated-measures ANOVA were
performed using IBM SPSS statistics 20 software. For Figs. 2, CF,
and 3, A and B, repeated-measurements data were analyzed using
mixed models, a generalization of repeated-measures ANOVA that
uses as many observations as are available for each specimen (rather
than requiring complete data from each specimen to be included in the
analysis). Mixed models expressed mean scores as functions of both
time and group membership, and mean scores at any particular time
point (in Figs. 2, C and D, and 3, A and B) were compared using
approximate t-tests corresponding to linear contrasts, as implemented
in the ESTIMATE statement of PROC MIXED in SAS. Areas under
the curve (as calculated by the trapezoidal rule) and mean scores at
any particular time point were compared using linear contrasts embedded in the mixed models. Figure 3, C and D, was also analyzed by
mixed models with treatment replacing time as the within-subjects
factor. Version 9.2 of SAS software was used for mixed-model
analyses. Nonrepeated measurement data were analyzed by Students
t-test (Fig. 4D) or Mann-Whitney rank sum test when the data failed
the Shapiro-Wilk normality test (Fig. 4, AC). These analyses were
completed using SigmaPlot 11.0 software.
RESULTS

Maternal body weight and food intake. After 7 days in

sedentary or exercise groups, female ICR mice were bred to
male mice for 10 days to ensure a maximum number of
pregnancies. During breeding, males also had access to the
running wheel. Depending on day of conception, exercise dams
had running wheel access for a minimum of 7 and maximum of
17 days prior to conception. Eleven of 18 exercise females
conceived on the first night of mating, whereas 14 of 18

AJP-Endocrinol Metab doi:10.1152/ajpendo.00213.2012 www.ajpendo.org

E1063

Running distance (km/day)

PERINATAL EXERCISE IMPROVES OFFSPRING HEALTH

14
12
10
8
6

Pups Born

2
0
0

10

20

30

40

Time (days)

Pups Born

55

Body weight (g)

50
45
40
35

30

Sedentary
Exercise

25
0
0

Time (weeks)

Food intake (g/day)

35
30
Sedentary
Exercise

25

Pups Born

20
15

10
5

** **

0
0

Time (weeks)
Fig. 1. Maternal exercise affected food intake but not body weight in female mice
that were set up in a breeding scheme. A: maternal mean run distance per day.
Arrows depict when males were present in the cages for breeding and when pups
were born. Exercise decreased as dams approached delivery, which is designated
as day 29. Data in B and C were not matched for day of delivery. Arrows indicate
when males were present for breeding and the range of days over which pups were
born. There were no differences in body weight between sedentary and exercised
dams (B), but there was a significant increase in food intake in pregnant exercise
dams compared with sedentary dams (C); n 18 for A, n 20 for sedentary, and
n 18 for exercise in B and C. Error bars indicate SE.

females conceived within the first 2 days of mating. Only four

exercise females included in the study conceived after the first
2 days of mating, meaning that the majority of female mice ran
only 7 8 days prior to conception. Offspring were weaned at
21 days of age and were placed into cages that did not contain
running wheels. Further details are provided in RESEARCH DESIGN AND METHODS.

Running data (Fig. 1A) were matched so that day 29 correlates to delivery day regardless of day of conception in relation
to being placed on the running wheels. Mean running distance
per day increased over the first 7 days as the female mice grew
accustomed to the running wheels (Fig. 1A). From days 8 17,
a male mouse was also present in the cage for breeding, and
running distance increased most likely due to male running.
Mean running distance decreased as the female mice approached delivery on day 29 and was maintained at lower
levels during lactation. Average running distance during nursing was significantly lower than average running distance prior
to pregnancy (P 0.001).
Figure 1, B and C, shows maternal body weight and food
intake. These data were not matched for day of delivery
because the values were only measured once/week. There were
no significant differences in body weight due to maternal
exercise prior to or during pregnancy and lactation (Fig. 1B).
Figure 1C shows the weekly food intake values divided by 7 as
a measurement of daily food intake. To determine food intake
while males were in the cage, food intake was divided by 2 to
account for two mice in the cage. There was a significant
increase in food intake prior to and during mating and pregnancy (P 0.001; repeated-measures ANOVA) in the running
dams at weeks 2, 3, and 4 (P 0.015, P 0.001, and P
0.001, respectively) (Fig. 1C). There were no differences in
food intake during nursing (weeks 57). Maternal running
during pregnancy did not significantly affect pregnancy rate or
litter size (Table 1). There were also no significant differences
in mean pup body weight per litter on postnatal days 7, 14, or
21 (Table 1).
Glucose and insulin tolerance in offspring. The offspring
born to sedentary and exercised dams (shown in Fig. 1, AC)
were then used for further analyses. There were no significant
differences in body weight in female or male offspring born to
sedentary or exercised dams from 3 to 76 wk of age (Fig. 2, A
and B). At 3132 wk of age, both female and male offspring
born to sedentary and exercised dams were fasted and given an
oral dose of glucose (2 g/kg body wt). Circulating blood
glucose values were measured after the oral glucose challenge
(data not shown). Overall glucose disposal was improved
significantly in female and male offspring born to exercised
dams compared with those from sedentary dams (P 0.023
and P 0.005, respectively). Oral glucose tolerance was again
measured in the offspring at 36 37 wk of age. Consistent with
earlier results, overall glucose disposal was improved in female
and male offspring born to exercised dams (P 0.004 and P
0.011, respectively). The female (Fig. 2C) and male (Fig. 2D)
Table 1. Effect of maternal exercise on pregnancy rates,
litter size, and pup body weights
Parameter

Sedentary (SE)

Exercise (SE)

Pregnancy rate
Subsequent analyses on litters (n)
Pups/litter
PND 7 pup body weight, g*
PND 14 pup body weight, g*
PND 21 pup body weight, g*

26/28
20
11.38 (0.71)
5.67 (0.09)
9.17 (0.20)
14.55 (0.19)

25/28
18
10.40 (0.74)
5.42 (0.15)
8.96 (0.25)
14.14 (0.40)

*Postnatal day (PND) pup body weights were calculated by averaging pup
body weights/litter. Each litter average was then used to calculate the mean pup
body weight/group.

AJP-Endocrinol Metab doi:10.1152/ajpendo.00213.2012 www.ajpendo.org

E1064

PERINATAL EXERCISE IMPROVES OFFSPRING HEALTH

70

Male Offspring

60

50
40
30
20

Sedentary
Exercise

10

50
40
30
20

Sedentary
Exercise

10

0
0

20

40

60

80

20

40

C 350

Female Offspring

Glucose (mg/dL)

Glucose (mg/dL)

300
250

**

200
150
100

Sedentary
Exercise

50

300

200

100

Sedentary
Exercise

0
0

30

60

90

120

**

25

60

35

120

30

25

20

90

Male Offspring

AUC (g/dL min)

Female Offspring

30

Time (min)

Time (min)
30

80

Male Offspring

400

60

Time (week)

Time (week)

AUC (g/dL min)

Fig. 2. Mature female and male Institute of

Cancer Research (ICR) offspring born to
exercised dams had improved glucose disposal independent of body weight differences. There were no significant differences
observed in female (A) or male offspring (B)
body weight. Following an oral glucose challenge, blood glucose levels were significantly reduced 30 min after glucose administration in female offspring (C) and 30 and
60 min after glucose administration in male
offspring (D) from exercised dams compared
with those from sedentary dams. Area under
the curve (AUC) of circulating blood glucose
was also significantly reduced in female (E)
and male offspring (F) from exercised dams.
*P 0.05 and **P 0.01 compared with
offspring born to sedentary dams; n 85 for
sedentary and 57 for exercise in A, and n
72 for sedentary and 84 for exercise in B.
Sample size in A and B represents the no. of
female and male offspring that were weaned
originally in week 3; n 19 for sedentary
and n 18 for exercise in C and E; n 18
for sedentary and n 18 for exercise in D
and F. Error bars indicate SE.

Female Offspring

Body weight (g)

Body weight (g)

A60

20

15

15

10

10

Sedentary

offspring born to exercised dams had significantly enhanced

glucose disposal after 30 min compared with offspring from
sedentary dams (P 0.001 and P 0.010, respectively). In
addition, male offspring born to exercised dams also had
significantly lower glucose levels 60 min after the glucose dose
(P 0.019). Area under the curve was decreased significantly
in female (Fig. 2E) offspring from exercised dams compared
with those from sedentary dams (P 0.009), with a similar
effect observed in male (Fig. 2F) offspring born to exercised
dams (P 0.026). Oral glucose tolerance was tested for a final
time in offspring at 7172 wk of age. Again, overall glucose
tolerance was improved significantly in female (P 0.032)
and male (P 0.039) offspring from exercised dams compared
with those born to sedentary dams (data not shown).
We then focused on the mechanism of enhanced glucose
disposal in female and male offspring born to exercised dams
by performing an insulin tolerance test. Blood glucose levels
drop over time in response to the exogenous insulin, and the
rate of disposal provides an index of insulin sensitivity. At
3334 wk of age mature female offspring were fasted, and
insulin at 0.75 IU/kg body wt was injected. Offspring from

Exercise

Sedentary

Exercise

both sedentary and exercised dams had consequential lowering

of blood glucose in response to insulin. Offspring born to
exercised dams had significantly improved overall glucose
disposal (P 0.024) compared with those from sedentary
dams, with significantly enhanced disposal at 15, 30, and 60
min postinjection (P 0.005, P 0.017, and P 0.035,
respectively), suggesting that these mice were more insulin
sensitive (Fig. 3A). Mature male offspring at 43 44 wk of age
were fasted and injected with a higher dose of insulin to ensure
glucose uptake (1.25 IU/kg body wt). Offspring born to exercised dams had overall improved glucose disposal (P 0.003),
with significantly enhanced disposal at 15, 30, 60, and 120 min
postinjection (P 0.001, P 0.010, P 0.036, and P
0.025, respectively) compared with offspring from sedentary
dams (Fig. 3B). These data confirmed that middle-aged offspring born to exercised dams had improved insulin sensitivity
compared with offspring born to sedentary dams.
2-Deoxyglucose uptake in offspring muscle and adipose.
Skeletal muscle and adipose tissue are responsible for the
majority of insulin-sensitive glucose uptake in vivo (36, 39).
We next set out to determine which tissues were responsible

AJP-Endocrinol Metab doi:10.1152/ajpendo.00213.2012 www.ajpendo.org

E1065

PERINATAL EXERCISE IMPROVES OFFSPRING HEALTH

B 140

Female Offspring

120

80

**

60

40

*
Sedentary
Exercise

20

100
80

30

60

90

120

150

180

Female Offspring

** # #
Sedentary
Exercise

30

60

90

120

150

180

Time (min)

**

Adipose 2-DG transport

(mol/g/h)

Muscle 2-DG transport

(mol/g/h)

Sedentary
Exercise
0

Time (min)

40

**

60

20

Male Offspring

120

100

Glucose (mg/dL)

Glucose (mg/dL)

Female Offspring

** # #
Sedentary
Exercise

Basal

+ 100 nM insulin

Basal

+ 100 nM insulin

Fig. 3. Mature female and male ICR offspring born to exercised dams show enhanced insulin sensitivity. Female and male offspring were fasted prior to an insulin
tolerance test. A: blood glucose levels in female offspring from exercised dams were significantly decreased at 15, 30, and 60 min after insulin injection compared
with offspring from sedentary dams. B: blood glucose levels in male offspring from exercised dams were significantly decreased at 15, 30, 60, and 120 min after
insulin injection compared with offspring from sedentary dams. C: both sedentary and exercise dam female offspring muscle showed significantly increased
2-deoxyglucose (2-DG) uptake in response to 100 nM insulin treatment compared with basal uptake. In addition, muscle from offspring born to exercised dams
showed significantly increased 2-DG uptake in response to insulin compared with offspring from sedentary dams. D: adipose from exercise dam offspring show
significantly increased 2-DG uptake in response to insulin treatment compared with basal uptake and compared with insulin-stimulated uptake in adipose from
offspring born to sedentary dams. *P 0.05 and **P 0.01 compared with offspring born to sedentary dams (A and B) or insulin-treated compared with basal
sedentary or exercise control (C and D). ##P 0.01 compared with insulin-treated sedentary; n 20 for sedentary and 18 for exercise in A; n 16 for sedentary
and 14 for exercise in B; n 6 for sedentary and exercise in C; n 4 for sedentary and exercise in D. Error bars indicate SE.

for the enhanced insulin sensitivity observed in the mature female

offspring born to exercised dams. Therefore, soleus muscle was
isolated, and 2-deoxyglucose (2-DG) uptake was measured in
the presence and absence of 100 nM insulin in vitro (Fig. 3C).
Muscle collected from offspring born to exercised dams
trended toward increased 2-DG uptake compared with muscle
from sedentary dam offspring in the basal state (P 0.151).
Muscle isolated from offspring born to both sedentary and
exercised dams showed significant increases in 2-DG uptake
in response to insulin compared with their basal uptake levels
(P 0.001 in both comparisons). Importantly, the muscles
from exercised dam offspring exhibited significantly greater
glucose uptake in response to the insulin dose compared with
muscle from sedentary dam offspring (P 0.007). A similar
design was used to measure 2-DG uptake into isolated adipose
in a separate cohort of female offspring born to sedentary and
exercised dams (Fig. 3D). The addition of 100 nM insulin to
adipose explants isolated from sedentary offspring did not
significantly affect 2-DG uptake (P 0.225), which suggests
a level of insulin resistance. However, adipose explants collected from offspring born to exercised dams were more
sensitive and had significantly increased 2-DG uptake in response to insulin compared with their basal state (P 0.002)
as well as compared with the insulin-treated explants isolated
from offspring born to sedentary dams (P 0.008). Both
skeletal muscle and adipose isolated from offspring born to

exercised dams exhibited enhanced insulin sensitivity. These

findings suggest that peripheral tissue insulin sensitivity is the
mechanism largely responsible for enhanced glucose disposal
following an oral glucose challenge.
Offspring body composition. Body composition was analyzed by quantitative magnetic resonance imaging (EchoMRI;
Echo Medical Systems) in mature 39-wk-old female and male
offspring. Fat and lean mass are expressed as a percent of body
weight. There were no differences observed in female offspring fat (Fig. 4A) or lean content (Fig. 4C). There were also
no differences in percent total water between female offspring
from exercised (41.1 1.2) or sedentary dams (42.3 1.2).
However, there were significant differences observed in male
offspring body composition. Figure 4B shows that male offspring from exercised dams had significantly reduced fat content compared with male offspring from sedentary dams (P
0.004). In addition, Fig. 4D shows that male offspring born to
exercised dams also had significantly increased lean tissue
composition compared with male offspring from sedentary
dams (P 0.001). Male offspring from exercised dams also
had significantly increased percent total water (53.9 0.5)
compared with male offspring from sedentary dams (51.2
0.8) (P 0.024). Offspring body composition was again
analyzed at 68 wk of age. Consistent with earlier analysis, there
were no significant differences in female offspring fat or lean
mass (data not shown). At this age, however, female offspring

AJP-Endocrinol Metab doi:10.1152/ajpendo.00213.2012 www.ajpendo.org

E1066

PERINATAL EXERCISE IMPROVES OFFSPRING HEALTH

50

Female Offspring

30

Male Offspring

**
Fat (%)

30
20

20

10

10
0

Sedentary

80

Exercise

Female Offspring

Sedentary

80

Exercise

Male Offspring

Lean tissue (%)

**
Lean tissue (%)

Fig. 4. Maternal exercise significantly impacts

body composition in male but not female offspring. Total body fat mass and lean mass were
analyzed in mature female and male offspring
using EchoMRI. Fat and lean mass results are
shown as %body weight. There was no difference
observed in female offspring %fat (A), but male
offspring born to exercised dams had significantly
lower %fat (B) compared with offspring from
sedentary dams. Female offspring %lean tissue
(C) was unchanged, whereas male %lean tissue
(D) was increased significantly in offspring born
to exercised dams compared with those born to
sedentary dams. **P 0.01 compared with offspring born to sedentary dams; n 20 for sedentary and 18 for exercise in A and C; n 19 for
sedentary and 14 for exercise in B and D. Data
were not normally distributed in AC; horizontal
line indicates median. Data were normally distributed in D; horizontal line indicates mean.

Fat (%)

40

60

40

20

60

40

20

Sedentary

from exercised dams had significantly increased percent total

water (P 0.047) compared with female offspring from
sedentary dams (data not shown). Again, there were significant
differences in male body composition. Males from exercised
dams had significantly reduced percent fat content (P 0.023)
and significantly increased percent lean tissue (P 0.040)
compared with males from sedentary dams (data not shown).
However, there were no differences in male offspring percent
total water at the older age (data not shown). It appears that
changes in body composition were not required for glucose
tolerance improvements observed in the female offspring born
to exercised dams but may play a role in changes in glucose
disposal observed in male offspring.
DISCUSSION

We have found that maternal exercise prior to and during

pregnancy and lactation can improve long-term metabolic
outcomes in offspring. Glucose disposal was significantly enhanced in both female and male offspring born to exercised
dams compared with those from sedentary dams. In addition,
male and female offspring from exercised dams were found to
be more sensitive to exogenous insulin. In female offspring, we
also found that excised skeletal muscle and fat pads from
offspring born to exercised dams were more sensitive to in
vitro insulin stimulation compared with those from sedentary
dams. Previously, exercise during pregnancy has been shown
to result in acute and long-term reduction in glucose that
reaches the fetus (6, 46). Changes in glucose availability during
development could have long-lasting effects in that mature
offspring become more sensitive to glucose changes. Consistent with our findings, Vanheest and Rodgers (47) used moderate speed treadmill running (20 m/min) in streptozotocininduced diabetic pregnant rats and found that their offspring
had improved glucose tolerance compared with offspring born
to sedentary diabetic and nondiabetic dams. Although the
connection is somewhat limited, this does suggest that the

Exercise

Sedentary

Exercise

improvement of glucose regulation by maternal exercise will

be observed in multiple species.
Previous studies by other laboratories have looked at different maternal and offspring outcomes resulting from exercise
during pregnancy. In human studies, maternal exercise effects
have focused mainly on pregnancies complicated by gestational diabetes. In a study of women with gestational diabetes,
it was found that resistance exercise was effective in lowering
the number of women who needed insulin therapy to maintain
normal blood glucose levels (9). In a study of healthy pregnant
women, mild physical activity was found to lower the risk of
developing gestational diabetes (26). In contrast, a recent
randomized control trial in women of normal body weight
showed that exercise during pregnancy did not decrease the
prevalence of gestational diabetes compared with sedentary
controls (43). Few human studies have investigated offspring
effects of maternal exercise during pregnancy. These studies
have focused mainly on birth weights, body composition, and
cognitive outcomes. In several studies, exercise during pregnancy reduced birth weights (7, 20). Lower birth weights were
found to coincide with reduced fat mass at birth as well as
reduced cord serum concentrations of growth hormones. At 5
yr of age, children from mothers who exercised during pregnancy were shown to have improved scores on general intelligence and oral language skills tests (7). Animal studies have
detected neurological changes in offspring as a result of exercise during pregnancy. Maternal swimming in rats was found
to increase hippocampal neurogenesis in rat pups that was then
associated with improved memory in the pups (25). A similar
study found that, in mice, maternal running during pregnancy
and nursing resulted in a 40% increase in total granule cells in
the offspring hippocampus (2). Recently, Herring et al. (19)
showed that short-term exercise during pregnancy was able to
reduce Alzheimer pathology in offspring in a transgenic mouse
model that is predisposed to the disease. Exercise during

AJP-Endocrinol Metab doi:10.1152/ajpendo.00213.2012 www.ajpendo.org

PERINATAL EXERCISE IMPROVES OFFSPRING HEALTH

pregnancy can clearly affect many maternal and offspring

outcomes.
Finding the mechanisms behind the observed metabolic
changes in offspring due to perinatal exercise will be an
important focus of future studies. This was beyond the scope of
the current study given the number of possible epigenetic and
sex-specific changes that are associated with developmental
programming models. Developmental programming of metabolism has been observed in human and animal models of
intrauterine growth restriction (IUGR). IUGR in humans and
animals is known to decrease glucose disposal and insulin
sensitivity in mature offspring (35, 40). Although the link
between IUGR and decreases in insulin sensitivity has been
known for decades, not until recently have researchers begun
to investigate the mechanisms behind these metabolic derangements. Studies have found decreases in the expression and
insulin-stimulated activation of many proteins involved in the
insulin-signaling pathway that correlate with decreases in
whole body insulin sensitivity (21, 32). Further complicating
the elucidation of the mechanism is that many of the metabolic
changes observed in developmental programming models do
not occur until offspring are aged. Because of this, it is unclear
whether offspring from sedentary dams develop a dysfunction
in insulin sensitivity as they age or whether offspring from
exercised dams are protected from a natural, age-related decline in insulin sensitivity. It is important to note that in the
current study glucose tolerance was first tested in 3-mo-old
offspring, but differences were not detected until the offspring
reached 7 mo of age.
The effects of developmental programming on long-term
offspring health have been shown to be sex specific in several
studies (13, 29). A similar observation was made in this study
when significant effects on the percentage of fat and lean tissue
in response to maternal exercise were found only in male
offspring despite an improvement in glucose tolerance in both
females and males. Perhaps more important than the change in
body composition in male offspring was the lack of changes in
fat and lean tissue in female offspring, highlighting improved
insulin sensitivity as the potential mechanism contributing to
enhanced glucose uptake in these offspring. It will be necessary to determine whether increased lean tissue plays a unique
role in improved glucose uptake in male offspring to elucidate
divergent mechanisms between males and females.
The findings from this study are an important step in discovering potential beneficial effects of maternal exercise for
offspring. Future studies will look at the mechanism through
which maternal exercise improves offspring glucose homeostasis, including investigating changes in milk content and
production during lactation. Experiments can be designed to
focus on periods of time before, during, and/or after pregnancy,
when maternal exercise is essential for beneficial effects in
offspring. Furthermore, cross-fostering strategies can be used
to control multiple factors.
One recent study showed that high-fat-fed male rats produced female offspring with impaired -cell function (29), and
another showed that low-protein diet consumption by male
mice prior to mating affected metabolic gene expression in
offspring (3). Therefore, it will be necessary to determine
whether paternal exercise prior to fertilization influences the
offspring outcomes observed in Figs. 2 4. One possible way to
control for paternal influence would be to use artificial insem-

E1067

ination. It is also important to address that the presence of a

running wheel in the cage may serve as enrichment for the
exercising dams. Although no studies have been conducted to
look at maternal environmental enrichment and its effects on
the metabolic health of offspring, a few animal studies have
shown that it can improve maternal and offspring cognitive
functions (24, 27, 42). It may be necessary in future studies to
control for this as a possible confounding factor by using a
controlled exercised paradigm in which there is no running
wheel in the home cage. Rather, mice would be removed from
the home cage daily and would exercise for a predetermined
amount of time using any one of a number of exercise paradigms. This design would also control for the vast amount of
running seen in the voluntary exercise paradigm.
Much work is left to be done, but our studies provide new
information on the positive impact that perinatal exercise can
have on offspring metabolic health in a mouse model. Such an
intervention provides a realistic mechanism to improve insulin
sensitivity in the next generation and positively impact insulinresistant states.
ACKNOWLEDGMENTS
We thank the Comparative Medicine Section (CMS) at the National
Institute on Aging (NIA), Dawn Boyer (CMS, NIA), Dawn Nines (CMS,
NIA), and Caitlin Younts (Laboratory of Experimental Gerontology, NIA) for
their help on an earlier pilot experiment at the NIA.
GRANTS
This study was supported by the National Institutes of Health [NIH;
NIAMS-AR-55246 to K. A. Esser, R01-EY-012998 to F. H. Andrade, R01DK-090460 to K. J. Pearson, and the research core of the COBRE on Obesity
and Cardiovascular Diseases from the National Center for Research Resources
(5P20-RR-021954-05) and the National Institute of General Medical Sciences
(8P20-GM-103527-05)]. L. G. Carter was supported by an NIH training grant
(DK-07778). This research was supported in part by the Intramural Research
Program of the NIA, NIH.
DISCLOSURES
No conflicts of interest, financial or otherwise, are declared by the authors.
AUTHOR CONTRIBUTIONS
L.G.C., K.N.L., R.d.C., and K.J.P. did the conception and design of the
research; L.G.C., D.C.W., C.M.T., S.Y.N.T., P.S., M.L.G.-C., and G.W.
performed the experiments; L.G.C., P.S., M.L.G.-C., F.H.A., R.J.C., and K.J.P.
analyzed the data; L.G.C., P.S., M.L.G.-C., R.J.C., K.A.E., J.M.E., R.d.C., and
K.J.P. interpreted the results of the experiments; L.G.C. and K.J.P. prepared
the figures; L.G.C., K.N.L., P.S., M.L.G.-C., J.M.E., R.d.C., and K.J.P. drafted
the manuscript; L.G.C., K.N.L., D.C.W., C.M.T., P.S., M.L.G.-C., G.W.,
F.H.A., K.A.E., J.M.E., R.d.C., and K.J.P. edited and revised the manuscript;
L.G.C., K.N.L., D.C.W., C.M.T., P.S., M.L.G.-C., G.W., F.H.A., R.J.C.,
K.A.E., J.M.E., R.d.C., and K.J.P. approved the final version of the manuscript.
REFERENCES
1. Baker JL, Olsen LW, Sorensen TI. Childhood body-mass index and the
risk of coronary heart disease in adulthood. N Engl J Med 357: 2329
2337, 2007.
2. Bick-Sander A, Steiner B, Wolf SA, Babu H, Kempermann G. Running in pregnancy transiently increases postnatal hippocampal neurogenesis in the offspring. Proc Natl Acad Sci USA 103: 38523857, 2006.
3. Carone BR, Fauquier L, Habib N, Shea JM, Hart CE, Li R, Bock C,
Li C, Gu H, Zamore PD, Meissner A, Weng Z, Hofmann HA,
Friedman N, Rando OJ. Paternally induced transgenerational environmental reprogramming of metabolic gene expression in mammals. Cell
143: 1084 1096, 2010.
4. Centers for Disease Control and Prevention. National Diabetes Fact
Sheet: General Information and National Estimates on Diabetes in the

AJP-Endocrinol Metab doi:10.1152/ajpendo.00213.2012 www.ajpendo.org

E1068

5.

6.
7.
8.
9.
10.

11.

12.
13.
14.
15.
16.
17.
18.
19.

20.

21.
22.

23.
24.
25.

26.

PERINATAL EXERCISE IMPROVES OFFSPRING HEALTH

United States, 2007. Atlanta, GA: US Department of Health and Human

Services, Centers for Disease Control and Prevention, 2008.
Chambers MA, Moylan JS, Smith JD, Goodyear LJ, Reid MB.
Stretch-stimulated glucose uptake in skeletal muscle is mediated by
reactive oxygen species and p38 MAP-kinase. J Physiol 587: 33633373,
2009.
Clapp JF 3rd. The effects of maternal exercise on fetal oxygenation and
feto-placental growth. Eur J Obstet Gynecol Reprod Biol 110, Suppl 1:
S80 S85, 2003.
Clapp JF 3rd. Morphometric and neurodevelopmental outcome at age
five years of the offspring of women who continued to exercise regularly
throughout pregnancy. J Pediatr 129: 856 863, 1996.
Cobrin M, Koski KG. Maternal dietary carbohydrate restriction and
mild-to-moderate exercise during pregnancy modify aspects of fetal development in rats. J Nutr 125: 16171627, 1995.
de Barros MC, Lopes MA, Francisco RP, Sapienza AD, Zugaib M.
Resistance exercise and glycemic control in women with gestational
diabetes mellitus. Am J Obstet Gynecol 203: 556 e551e556, 2010.
Dyck R, Klomp H, Tan LK, Turnell RW, Boctor MA. A comparison of
rates, risk factors, and outcomes of gestational diabetes between aboriginal
and non-aboriginal women in the Saskatoon health district. Diabetes Care
25: 487493, 2002.
Fernandez-Twinn DS, Wayman A, Ekizoglou S, Martin MS, Hales
CN, Ozanne SE. Maternal protein restriction leads to hyperinsulinemia
and reduced insulin-signaling protein expression in 21-mo-old female rat
offspring. Am J Physiol Regul Integr Comp Physiol 288: R368 R373,
2005.
Gardner DS, Buttery PJ, Daniel Z, Symonds ME. Factors affecting
birth weight in sheep: maternal environment. Reproduction 133: 297307,
2007.
Gilbert JS, Nijland MJ. Sex differences in the developmental origins of
hypertension and cardiorenal disease. Am J Physiol Regul Integr Comp
Physiol 295: R1941R1952, 2008.
Gluckman PD, Hanson MA. Developmental and epigenetic pathways to
obesity: an evolutionary-developmental perspective. Int J Obes (Lond) 32,
Suppl 7: S62S71, 2008.
Gluckman PD, Hanson MA. Developmental origins of disease paradigm:
a mechanistic and evolutionary perspective. Pediatr Res 56: 311317,
2004.
Gluckman PD, Hanson MA, Beedle AS, Raubenheimer D. Fetal and
neonatal pathways to obesity. Front Horm Res 36: 6172, 2008.
Gluckman PD, Hanson MA, Pinal C. The developmental origins of adult
disease. Matern Child Nutr 1: 130 141, 2005.
Hales CN, Barker DJ. Type 2 (non-insulin-dependent) diabetes mellitus:
the thrifty phenotype hypothesis. Diabetologia 35: 595601, 1992.
Herring A, Donath A, Yarmolenko M, Uslar E, Conzen C, Kanakis D,
Bosma C, Worm K, Paulus W, Keyvani K. Exercise during pregnancy
mitigates Alzheimer-like pathology in mouse offspring. FASEB J 26:
117128, 2012.
Hopkins SA, Baldi JC, Cutfield WS, McCowan L, Hofman PL.
Exercise training in pregnancy reduces offspring size without changes in
maternal insulin sensitivity. J Clin Endocrinol Metab 95: 2080 2088,
2010.
Jensen CB, Martin-Gronert MS, Storgaard H, Madsbad S, Vaag A,
Ozanne SE. Altered PI3-kinase/Akt signalling in skeletal muscle of young
men with low birth weight. Plos One 3: e3738, 2008.
Jones HN, Woollett LA, Barbour N, Prasad PD, Powell TL, Jansson
T. High-fat diet before and during pregnancy causes marked up-regulation
of placental nutrient transport and fetal overgrowth in C57/BL6 mice.
FASEB J 23: 271278, 2009.
Kim H, Lee SH, Kim SS, Yoo JH, Kim CJ. The influence of maternal
treadmill running during pregnancy on short-term memory and hippocampal cell survival in rat pups. Int J Dev Neurosci 25: 243249, 2007.
Kiyono S, Seo ML, Shibagaki M, Inouye M. Facilitative effects of
maternal environmental enrichment on maze learning in rat offspring.
Physiol Behav 34: 431435, 1985.
Lee HH, Kim H, Lee JW, Kim YS, Yang HY, Chang HK, Lee TH,
Shin MC, Lee MH, Shin MS, Park S, Baek S, Kim CJ. Maternal
swimming during pregnancy enhances short-term memory and neurogenesis in the hippocampus of rat pups. Brain Dev 28: 147154, 2006.
Liu J, Laditka JN, Mayer-Davis EJ, Pate RR. Does physical activity
during pregnancy reduce the risk of gestational diabetes among previously
inactive women? Birth 35: 188 195, 2008.

27. Maruoka T, Kodomari I, Yamauchi R, Wada E, Wada K. Maternal

enrichment affects prenatal hippocampal proliferation and open-field behaviors in female offspring mice. Neurosci Lett 454: 28 32, 2009.
28. Mottola MF. The role of exercise in the prevention and treatment of
gestational diabetes mellitus. Curr Diab Rep 8: 299 304, 2008.
29. Ng SF, Lin RC, Laybutt DR, Barres R, Owens JA, Morris MJ. Chronic
high-fat diet in fathers programs beta-cell dysfunction in female rat
offspring. Nature 467: 963966, 2010.
30. Ogden CL, Carroll MD, Curtin LR, McDowell MA, Tabak CJ, Flegal
KM. Prevalence of overweight and obesity in the United States, 1999
2004. JAMA 295: 1549 1555, 2006.
31. Ozanne SE, Hales CN. Lifespan: catch-up growth and obesity in male
mice. Nature 427: 411412, 2004.
32. Ozanne SE, Jensen CB, Tingey KJ, Storgaard H, Madsbad S, Vaag
AA. Low birthweight is associated with specific changes in muscle
insulin-signalling protein expression. Diabetologia 48: 547552, 2005.
33. Paffenbarger RS Jr, Blair SN, Lee IM. A history of physical activity,
cardiovascular health and longevity: the scientific contributions of Jeremy
N Morris, DSc, DPH, FRCP. Int J Epidemiol 30: 1184 1192, 2001.
34. Petry CJ, Jennings BJ, James LA, Hales CN, Ozanne SE. Suckling a
protein-restricted rat dam leads to diminished albuminuria in her male
offspring in adult life: a longitudinal study. BMC Nephrol 7: 14, 2006.
35. Ravelli AC, van der Meulen JH, Michels RP, Osmond C, Barker DJ,
Hales CN, Bleker OP. Glucose tolerance in adults after prenatal exposure
to famine. Lancet 351: 173177, 1998.
36. Reaven GM. Banting lecture 1988. Role of insulin resistance in human
disease. Diabetes 37: 15951607, 1988.
37. Rogers PM, Mashtalir N, Rathod MA, Dubuisson O, Wang Z, Dasuri
K, Babin S, Gupta A, Markward N, Cefalu WT, Dhurandhar NV.
Metabolically favorable remodeling of human adipose tissue by human
adenovirus type 36. Diabetes 57: 23212331, 2008.
38. Samuelsson AM, Matthews PA, Argenton M, Christie MR, McConnell
JM, Jansen EH, Piersma AH, Ozanne SE, Twinn DF, Remacle C,
Rowlerson A, Poston L, Taylor PD. Diet-induced obesity in female mice
leads to offspring hyperphagia, adiposity, hypertension, and insulin resistance: a novel murine model of developmental programming. Hypertension 51: 383392, 2008.
39. Shepherd PR, Kahn BB. Glucose transporters and insulin action
implications for insulin resistance and diabetes mellitus. N Engl J Med
341: 248 257, 1999.
40. Simmons RA, Templeton LJ, Gertz SJ. Intrauterine growth retardation
leads to the development of type 2 diabetes in the rat. Diabetes 50:
2279 2286, 2001.
41. Snapp CA, Donaldson SK. Gestational diabetes mellitus: physical exercise and health outcomes. Biol Res Nurs 10: 145155, 2008.
42. Sparling JE, Mahoney M, Baker S, Bielajew C. The effects of gestational and postpartum environmental enrichment on the mother rat: A
preliminary investigation. Behav Brain Res 208: 213223, 2010.
43. Stafne SN, Salvesen KA, Romundstad PR, Eggebo TM, Carlsen SM,
Morkved S. Regular exercise during pregnancy to prevent gestational
diabetes: a randomized controlled trial. Obstet Gynecol 119: 29 36, 2012.
44. Tamashiro KL, Terrillion CE, Hyun J, Koenig JI, Moran TH. Prenatal
stress or high-fat diet increases susceptibility to diet-induced obesity in rat
offspring. Diabetes 58: 1116 1125, 2009.
45. Tarry-Adkins JL, Joles JA, Chen JH, Martin-Gronert MS, van der
Giezen DM, Goldschmeding R, Hales CN, Ozanne SE. Protein restriction in lactation confers nephroprotective effects in the male rat and is
associated with increased antioxidant expression. Am J Physiol Regul
Integr Comp Physiol 293: R1259 R1266, 2007.
46. Treadway JL, Young JC. Decreased glucose uptake in the fetus after
maternal exercise. Med Sci Sports Exerc 21: 140 145, 1989.
47. Vanheest JL, Rodgers CD. Effects of exercise in diabetic rats before and
during gestation on maternal and neonatal outcomes. Am J Physiol
Endocrinol Metab 273: E727E733, 1997.
48. White CL, Purpera MN, Morrison CD. Maternal obesity is necessary
for programming effect of high-fat diet on offspring. Am J Physiol Regul
Integr Comp Physiol 296: R1464 R1472, 2009.
49. Zhang C, Solomon CG, Manson JE, Hu FB. A prospective study of
pregravid physical activity and sedentary behaviors in relation to the risk
for gestational diabetes mellitus. Arch Intern Med 166: 543548, 2006.

AJP-Endocrinol Metab doi:10.1152/ajpendo.00213.2012 www.ajpendo.org