Embed Size (px)
Citation preview
Maternal Obesity and the Developmental Programming of Offspring Appetite and obesity: A role for leptin?
Paul D Taylor, Anne-Maj Samuelsson, Clive W Coen and Lucilla Poston.
Women’s Health Academic Centre
Note: for non-commercial purposes only
Maternal Obesity
Glucose, insulin, leptin, lipids, inflammatory response
Fetal macrosomia, increased adiposity
Persistently altered energy balance
Childhood and Adulthood Obesity
Transgenerational ‘Acceleration ‘ of Obesity?
Maternal pre-pregnancy BMI and gestational weight gain Risk of offspring obesity: Hochner 2012 Jerusalem study
• Adjusted means of offspring obesity outcomes at age 32 by quartiles of maternal pp BMI and GWG
• Greater mppBMI, independently of GWG, was significantly associated with higher offspring BMI, and WC.
• The observed associations were independent of characteristics reflecting the pre- peri- and post-natal environment, including current measures of SES and lifestyle.
• A similar associations of mppBMI and GWG for DBP and SBP was observed, but this disappeared after controlling for offspring current BMI.
• Mice, rats, and humans share all but 1% of each other's genes
• Reduce possible genetic influence
• Relatively quick life cycle
• Environment can be tightly controlled
• Enable different diets to be tested
• Investigate critical periods in development
Why use animal models to study mechanisms of Developmental Programming?
“Future studies that explore mechanisms underlying the intergenerational cycle of obesity are warranted to identify potentially novel targets for cardiometabolic risk-reduction interventions”.
Hochner et al 2012
Breeding Offspring of Obese Rats and Mice (OffOb)
6 week
Mating
Gestation & Lactation
6 week Gestation & Lactation
DAMS OFFSPRING
Control Diet Highly Palatable Obesogenic Diet: Pellets with 20% FAT + Sweetened Condensed Milk
Characterization of pre-Obese Phenotype
Day 30 Day 90
Weaning
Characterization of Obese Phenotype
Maternal Characteristics Page 5
B A
-5 -4 -3 -2 -1 0 2 4 6 8 10 12 14 16 18 20
Bod
y w
eigh
t (g)
10
20
30
40
50* **
Time (Gestational days)Time (weeks)-5 -4 -3 -2 -1 0 2 4 6 8 10 12 14 16 18 20
Bod
y w
eigh
t (g)
10
20
30
40
50* **
Time (Gestational days)Time (weeks)C
alor
ific
inta
ke (k
cal a
nim
al-1
day-
1 )
0
10
20
30
40
* *
-5 -4 -3 -2 -1 0 2 4 6 8 10 12 14 16 18 20Time (Gestational days)Time (weeks)
Cal
orifi
c in
take
(kca
l ani
mal
-1da
y-1 )
0
10
20
30
40
* *
-5 -4 -3 -2 -1 0 2 4 6 8 10 12 14 16 18 20Time (Gestational days)Time (weeks)
Control Dam Obese Dam
E
9.83.6Fat mass (g)34.414.8% Fat
18.820.5Lean mass (g)OBESE CONTROL
9.83.6Fat mass (g)34.414.8% Fat
18.820.5Lean mass (g)OBESE CONTROL
C
BA
T m
ass
(g)
0
0.1
0.2
0.3
*
Control Obese
BA
T m
ass
(g)
0
0.1
0.2
0.3
*
Control Obese
WA
T m
ass
(g)
01234567
Control Obese
***
WA
T m
ass
(g)
01234567
Control Obese
WA
T m
ass
(g)
01234567
Control Obese
***
D
Maternal Characteristics Page 6
Weaning
0
1
2
3
4
5
6
Triglyceride(mmol/L)
Cholesterol(mmol/L)
Glucose(10*mmol/L)
Insulin(100*pmol/L)
Leptin(100*pg/L)
Conc
entra
tions
*
***
0
1
2
3
4
5
6
Triglyceride(mmol/L)
Cholesterol(mmol/L)
Glucose(10*mmol/L)
Insulin(100*pmol/L)
Leptin(100*pg/L)
Conc
entra
tions
*
***
0
1
2
3
4
5
6
Triglyceride(mmol/L)
Cholesterol(mmol/L)
Glucose(10*mmol/L)
Insulin(100*pmol/L)
Leptin(10*pg/L)
Conc
entra
tions
***** ***
**
0
1
2
3
4
5
6
Triglyceride(mmol/L)
Cholesterol(mmol/L)
Glucose(10*mmol/L)
Insulin(100*pmol/L)
Leptin(10*pg/L)
Conc
entra
tions
***** ***
**
Late gestation
Offspring Phenotype: hyperphagia and adult obesity Page 7
MALE FEMALE
Time (weeks)0 5 10 15 20 25
body
wei
ght (
g)
0
5
10
15
20
25
30
35 *
Time (weeks)0 5 10 15 20 25
body
wei
ght (
g)
0
5
10
15
20
25
30
35 *
0 5 10 15 20 250
5
10
15
20
25
30
35*
Time (weeks)0 5 10 15 20 25
0
5
10
15
20
25
30
35*
Time (weeks)
Time (weeks)2 4 6 8 10 12 14 16 18 20 22
Cal
orifi
c in
take
(k
cal a
nim
al-1
wee
k-1)
40
60
80
100
120
Time (weeks)2 4 6 8 10 12 14 16 18 20 22
40
60
80
100
120* *
OC OO
Fat m
ass
(mg/
g B
W)
10
12
14
16
18
20
22***
OC OO
Fat m
ass
(mg/
g B
W)
6
8
10
12
14
16
**
0
1
2
3
4
5
6
Triglyceride(mmol/L)
Cholesterol(mmol/L)
Glucose(10*mmol/L)
Insulin(100*pmol/L)
Leptin (pg/L)
Con
cent
ratio
ns
***
**
*
0
1
2
3
4
5
6
Triglyceride(mmol/L)
Cholesterol(mmol/L)
Glucose(10*mmol/L)
Insulin(100*pmol/L)
Leptin (pg/L)
Con
cent
ratio
ns
* ***
*
MALE FEMALE
Time (weeks)0 5 10 15 20 25
body
wei
ght (
g)
0
5
10
15
20
25
30
35 *
Time (weeks)0 5 10 15 20 25
body
wei
ght (
g)
0
5
10
15
20
25
30
35 *
0 5 10 15 20 250
5
10
15
20
25
30
35*
Time (weeks)0 5 10 15 20 25
0
5
10
15
20
25
30
35*
Time (weeks)
Time (weeks)2 4 6 8 10 12 14 16 18 20 22
Cal
orifi
c in
take
(k
cal a
nim
al-1
wee
k-1)
40
60
80
100
120
Time (weeks)2 4 6 8 10 12 14 16 18 20 22
40
60
80
100
120
Time (weeks)2 4 6 8 10 12 14 16 18 20 22
Cal
orifi
c in
take
(k
cal a
nim
al-1
wee
k-1)
40
60
80
100
120
Time (weeks)2 4 6 8 10 12 14 16 18 20 22
40
60
80
100
120* *
OC OO
Fat m
ass
(mg/
g B
W)
10
12
14
16
18
20
22***
OC OO
Fat m
ass
(mg/
g B
W)
6
8
10
12
14
16
**
0
1
2
3
4
5
6
Triglyceride(mmol/L)
Cholesterol(mmol/L)
Glucose(10*mmol/L)
Insulin(100*pmol/L)
Leptin (pg/L)
Con
cent
ratio
ns
***
**
*
0
1
2
3
4
5
6
Triglyceride(mmol/L)
Cholesterol(mmol/L)
Glucose(10*mmol/L)
Insulin(100*pmol/L)
Leptin (pg/L)
Con
cent
ratio
ns
* ***
*
0
1
2
3
4
5
6
Triglyceride(mmol/L)
Cholesterol(mmol/L)
Glucose(10*mmol/L)
Insulin(100*pmol/L)
Leptin (pg/L)
Con
cent
ratio
ns
***
**
*
0
1
2
3
4
5
6
Triglyceride(mmol/L)
Cholesterol(mmol/L)
Glucose(10*mmol/L)
Insulin(100*pmol/L)
Leptin (pg/L)
Con
cent
ratio
ns
* ***
*
A
B
C
D
Maternal Phenotype
Increased birthweight & weight at weaning (milk content) Hyperphagic and hypertensive from weaning Abdominal obesity, hyperleptinaemia, dyslipidaemia, hyperinsulinaemia
and reduced muscle mass by 3 months. Hyperglycaemia (type 2 DM) and fatty liver disease (NAFLD) by 6 months.
Offspring Phenotype
High-fat high-sugar diet leads to hypercalorific intake & maternal obesity. Hyperleptinaemia, hyperinsulinaemia in pregnancy Further hyperglycaemia, dyslipidaemia during suckling.
Summary (1): Maternal Obesity in rodents
Neonatal serum leptin and insulin in OffOb
Figure 1. Neonatal serum leptin and insulin concentrations in offspringof control and obese dams. Serum leptin (A) and insulin (B) were measuredin offspring of control dams (open bars) and obese dams (closed bars) overthe suckling period. * p<0.05, ** p<0.01 and *** p<0.01 versus offspring ofcontrol dams for the same period (n = 3-6). For longitudinal comparisons, asignificant difference (p<0.05) from the preceding period is indicated by # foroffspring of control dams and by † for offspring of obese dams.
0
1
2
3
4
5
6
7
2 7 8 9 11 13 14 15
Postnatal Day
Seru
m Ins
ulin (
ng/m
l)
OffConOffOb
**
** ***
* * **
Figure 4. Neonatal serum insulin concentrations. Serum insulin was measured by ELISA in offspring of control (OffCon) and obese dams (OffOb) at PD 2,7,8,9,11,13,14, and 15. ***P<0.001, **P<0.01, *P<0.05 vscontrol, t-test, n=4-11.
0
1
2
3
4
5
6
7
2 7 8 9 11 13 14 15
Postnatal Day
Seru
m Ins
ulin (
ng/m
l)
OffConOffOb
**
** ***
* * **
Figure 4. Neonatal serum insulin concentrations. Serum insulin was measured by ELISA in offspring of control (OffCon) and obese dams (OffOb) at PD 2,7,8,9,11,13,14, and 15. ***P<0.001, **P<0.01, *P<0.05 vscontrol, t-test, n=4-11.
2 7 8 9 11 13 14 15 180
10
20
30
40
50
Offspring of Obese DamsOffspring of Control Dams
********* ***
***
****
Post natal Day
Pla
sma
Lept
inC
once
ntra
tion
(ng/
ml)
2 7 8 9 11 13 14 15 180
10
20
30
40
50
Offspring of Obese DamsOffspring of Control Dams
********* ***
***
****
Post natal Day
Pla
sma
Lept
inC
once
ntra
tion
(ng/
ml)
A
Ser
um le
ptin
(ng
/ml)
***
************
***
*
B
012345
67
2-8 9-11 13-18
Lept
in m
RN
A e
xpre
ssio
n(C
opy
# pe
r ge
omea
nx
1000
)
**
Postnatal day
†
†
†
†
# #
# #
#
#
†
2 7 8 9 11 13 14 15 180
10
20
30
40
50
Offspring of Obese DamsOffspring of Control Dams
********* ***
***
****
Post natal Day
Pla
sma
Lept
inC
once
ntra
tion
(ng/
ml)
2 7 8 9 11 13 14 15 180
10
20
30
40
50
Offspring of Obese DamsOffspring of Control Dams
********* ***
***
****
Post natal Day
Pla
sma
Lept
inC
once
ntra
tion
(ng/
ml)
A
Ser
um le
ptin
(ng
/ml)
***
************
***
*
A
Ser
um le
ptin
(ng
/ml)
Ser
um le
ptin
(ng
/ml)
***
************
***
*
B
012345
67
2-8 9-11 13-18
Lept
in m
RN
A e
xpre
ssio
n(C
opy
# pe
r ge
omea
nx
1000
)
**
Postnatal day
B
012345
67
2-8 9-11 13-18
Lept
in m
RN
A e
xpre
ssio
n(C
opy
# pe
r ge
omea
nx
1000
)
**
Postnatal day
012345
67
2-8 9-11 13-18
Lept
in m
RN
A e
xpre
ssio
n(C
opy
# pe
r ge
omea
nx
1000
)
**
Postnatal day
†
†
†
†
# #
# #
#
#
†
Postnatal Day
Postnatal Day
Ser
um L
eptin
(ng/
ml)
Ser
um In
sulin
(ng/
ml)
†
0
1
2
3
4
5
6
2 7 8 9 11 13 14 15
A
B
Fig 2. Leptin mRNA expression in adipose tissue from inoffspring from control (open bars) and obese dams (closedbars). Day 2-8 (n=5-10), 9-11 (n=3-4), 13-18 (n=4-8) mRNAexpressed as copy number divided by the geometric meanof 2 reference genes (28S, Actin beta), multiplied by 1000.Days 13-18, **P<0.005 vs control.
0.0
1.0
2.0
3.0
4.0
5.0
6.0
7.0
2-8 9-11 13-18
lep
tin
mR
NA
exp
ress
ion
(Co
py
#/g
eo
mea
n*1
000) **
Post-natal day
Leptin in Developmental Programming
• Postnatal leptin surge in rodents (Ahima 1998) – a key developmental signal for the hypothalamus
• Leptin in early life modulates the formation of neural energy-regulation circuits in the hypothalamus (Bouret et al 2004).
• Maternal undernutrition can influences the timing and amplitude of the leptin surge (Delahaye et al 2008; Yura et al 2005; Stocker et al 2007; Attig et al 2008).
• Genetic models of obesity?
Exogenous leptin rescues arcuate nucleus development in neonatal but not adult Lepob/Lepob mice (Bouret et al, Science 2004)
Leptin in the neuro-development of the brain
Leptin Challenge in Day 30 Offspring 12
05
10152025303540
OffCon
24 H
our F
ood
Inta
ke (g
)
LeptinSaline
***
OffOb
0
5
10
15
20
25
OffOb
Cha
nge
in B
ody
Wei
ght (
g)
OffCon
*
LeptinSaline
05
1015
2025
303540
OffOb
24 H
our F
ood
Inta
ke (g
)
OffCon
**
LeptinSaline
0
2
4
6
8
10
12
14
16
OffObOffCon
*
Cha
nge
in B
ody
Wei
ght (
g) LeptinSaline
Males Females
24 hr Food Intake
Δ Body WeightMales Females
05
10152025303540
OffCon
24 H
our F
ood
Inta
ke (g
)
LeptinSalineLeptinSaline
******
OffOb
0
5
10
15
20
25
OffOb
Cha
nge
in B
ody
Wei
ght (
g)
OffCon
***
LeptinSalineLeptinSaline
05
1015
2025
303540
OffOb
24 H
our F
ood
Inta
ke (g
)
OffCon
******
LeptinSalineLeptinSaline
0
2
4
6
8
10
12
14
16
OffObOffCon
****
Cha
nge
in B
ody
Wei
ght (
g) LeptinSalineLeptinSaline
Males Females
24 hr Food Intake
Δ Body WeightMales Females
A role for the melanocortin system in programming of appetite?
From Ozanne et al, Clinical Science 2005
Leptin Signalling in Day 30 Offspring
Bregma Group Mean SEM n p
VMH -2.30 mm OffCon 339 36 6
" OffOb 304 71 4 NS
VMHdl -3.30 mm OffCon 1146 121 8
" OffOb 1152 288 4 NS
VMHvl -3.30 mm OffCon 765 50 8
" OffOb 789 185 5 NS
OffObOffCon
ARC pSTAT3-ir Cells ~3.30 mm Caudal to Bregma ARC pSTAT3-ir Cells
VMH pSTAT3-ir Cells
0
500
1000
1500
2000
2500
3000
-2.30 -3.30 -4.16 Num
ber o
f pS
TAT3
-ir C
ells
/mm
2
*
Bregma (mm)
Bregma Group Mean SEM n p
VMH -2.30 mm OffCon 339 36 6
" OffOb 304 71 4 NS
VMHdl -3.30 mm OffCon 1146 121 8
" OffOb 1152 288 4 NS
VMHvl -3.30 mm OffCon 765 50 8
" OffOb 789 185 5 NS
OffObOffCon
ARC pSTAT3-ir Cells ~3.30 mm Caudal to Bregma ARC pSTAT3-ir Cells
VMH pSTAT3-ir Cells
0
500
1000
1500
2000
2500
3000
-2.30 -3.30 -4.16 Num
ber o
f pS
TAT3
-ir C
ells
/mm
2
*
Bregma (mm)
AgRP Immunoreactivity in Female PVH at Day 30
AgRP Fluorescence
**0.05
0.1
0.15
0.2
0.25
0.3
Bregma -1.80 mm
AgR
P-Im
mun
orea
ctiv
ity(%
Are
a PV
H)
OffObOffCon
AgRP Fluorescence (Bregma -1.80 mm) AgRP Fluorescence
**0.05
0.1
0.15
0.2
0.25
0.3
Bregma -1.80 mm
AgR
P-Im
mun
orea
ctiv
ity(%
Are
a PV
H)
**0.05
0.1
0.15
0.2
0.25
0.3
Bregma -1.80 mm
AgR
P-Im
mun
orea
ctiv
ity(%
Are
a PV
H)
OffObOffCon OffObOffCon
AgRP Fluorescence (Bregma -1.80 mm)
Summary (2)
• Maternal obesity has adverse consequences for offspring energy balance including leptin resistance and hyperphagia.
• We hypothesise that leptin resistance in the ARC is acquired during a critical period due to an amplified and prolonged neonatal leptin surge.
• Deficits in leptin-signalling may impair leptin’s neurotrophic effects on AgRP projections to the PVH.
• Abnormal neuronal development in the appetite regulatory centres of the brain may permanently programme hyperphagia and therefore obesity in adult life.
Leptin Treatment in naïve rat pups Postnatal Day 9-15
Treatment PD9-15
PD2 PD7 PD8 PD9 PD11 PD200.0
0.1
0.2
0.3Saline
Leptin
** **
Postnatal Day
Hea
rt w
eigh
t (g)
Treatment PD9-15
PD2 PD7 PD8 PD9 PD11 PD12 PD14 PD200
20
40
60
80
100
Seru
m L
eptin
(ng/
ml)
******
*** ***leptinsaline
Postnatal Day
Neonatal serum leptin concentrations achieved leptin (3mg/kg ip) and saline treated pups (male and female combined) ***P<0.001, **P<0.01 versus saline using t-test, n=4-7.
Figure 1. Neonatal serum leptin and insulin concentrations in offspringof control and obese dams. Serum leptin (A) and insulin (B) were measuredin offspring of control dams (open bars) and obese dams (closed bars) overthe suckling period. * p<0.05, ** p<0.01 and *** p<0.01 versus offspring ofcontrol dams for the same period (n = 3-6). For longitudinal comparisons, asignificant difference (p<0.05) from the preceding period is indicated by # foroffspring of control dams and by † for offspring of obese dams.
0
1
2
3
4
5
6
7
2 7 8 9 11 13 14 15
Postnatal Day
Seru
m Ins
ulin (
ng/m
l)
OffConOffOb
**
** ***
* * **
Figure 4. Neonatal serum insulin concentrations. Serum insulin was measured by ELISA in offspring of control (OffCon) and obese dams (OffOb) at PD 2,7,8,9,11,13,14, and 15. ***P<0.001, **P<0.01, *P<0.05 vscontrol, t-test, n=4-11.
0
1
2
3
4
5
6
7
2 7 8 9 11 13 14 15
Postnatal Day
Seru
m Ins
ulin (
ng/m
l)
OffConOffOb
**
** ***
* * **
Figure 4. Neonatal serum insulin concentrations. Serum insulin was measured by ELISA in offspring of control (OffCon) and obese dams (OffOb) at PD 2,7,8,9,11,13,14, and 15. ***P<0.001, **P<0.01, *P<0.05 vscontrol, t-test, n=4-11.
2 7 8 9 11 13 14 15 180
10
20
30
40
50
Offspring of Obese DamsOffspring of Control Dams
********* ***
***
****
Post natal Day
Pla
sma
Lept
inC
once
ntra
tion
(ng/
ml)
2 7 8 9 11 13 14 15 180
10
20
30
40
50
Offspring of Obese DamsOffspring of Control Dams
********* ***
***
****
Post natal Day
Pla
sma
Lept
inC
once
ntra
tion
(ng/
ml)
A
Ser
um le
ptin
(ng
/ml)
***
************
***
*
B
012345
67
2-8 9-11 13-18
Lept
in m
RN
A e
xpre
ssio
n(C
opy
# pe
r ge
omea
nx
1000
)
**
Postnatal day
†
†
†
†
# #
# #
#
#
†
2 7 8 9 11 13 14 15 180
10
20
30
40
50
Offspring of Obese DamsOffspring of Control Dams
********* ***
***
****
Post natal Day
Pla
sma
Lept
inC
once
ntra
tion
(ng/
ml)
2 7 8 9 11 13 14 15 180
10
20
30
40
50
Offspring of Obese DamsOffspring of Control Dams
********* ***
***
****
Post natal Day
Pla
sma
Lept
inC
once
ntra
tion
(ng/
ml)
A
Ser
um le
ptin
(ng
/ml)
***
************
***
*
A
Ser
um le
ptin
(ng
/ml)
Ser
um le
ptin
(ng
/ml)
***
************
***
*
B
012345
67
2-8 9-11 13-18
Lept
in m
RN
A e
xpre
ssio
n(C
opy
# pe
r ge
omea
nx
1000
)
**
Postnatal day
B
012345
67
2-8 9-11 13-18
Lept
in m
RN
A e
xpre
ssio
n(C
opy
# pe
r ge
omea
nx
1000
)
**
Postnatal day
012345
67
2-8 9-11 13-18
Lept
in m
RN
A e
xpre
ssio
n(C
opy
# pe
r ge
omea
nx
1000
)
**
Postnatal day
†
†
†
†
# #
# #
#
#
†
Postnatal Day
Postnatal Day
Ser
um L
eptin
(ng/
ml)
Ser
um In
sulin
(ng/
ml)
†
0
1
2
3
4
5
6
2 7 8 9 11 13 14 15
A
B
Figure S2. Weight gain and food intake (24hr) following a bolus leptin challenge (10 mg/kg i.p.) in fasted saline-treated (S-Tx) or leptin-treated (L-Tx) male and female rats at (A) 30 days and (B) 5 months of age. ***P<0.001, **P<0.01, *P<0.05 versus saline injection using t-test, n=8. Abbreviation S-Tx-saline treated; L-Tx-leptin treated
*
0
5
10
15
20 Fo
od in
take
(g)
S-Tx L-Tx
***
0
5
10
15
Wei
ght g
ain
(g)
S-Tx L-Tx
Saline injection Leptin bolus injection
Male
S-Lx L-Tx 0
5
10
15
20
25
Wei
ght g
ain
(g)
*
Female Male Female
Male
**
S-Tx L-Tx 0
10
20
30
40
Food
inta
ke (g
)
Female Male
S-Tx L-Tx 0
5
10
15
Wei
ght g
ain
(g)
**
0
5
10
15
20
Food
inta
ke (g
)
S-Tx L-Tx
*
Female
0
5
10
15
20
25
Wei
ght g
ain
(g)
S-Tx L-Tx S-Tx L-Tx 0
10
20
30
40
Food
inta
ke (g
)
** *
Leptin Challenge in L-Tx neonates at 30 days and 5 months
Body weight and organ weights 5 and 12 months
5 months 12 months
Weight male female male female
S-Tx L-Tx S-Tx L-Tx S-Tx L-Tx S-Tx L-Tx
Bodyweight
(g)
222.9±6.9 238.2±12.1 188.5±10.9 194.9±9.1 600.0±29.6 631.3±23.7 346.8±7.2 402.2±17.2†
Inguinal WAT
(mg)
1.32±0.15 2.03±0.35* 1.01±0.15 1.86±0.05† 5.31±0.41 9.25±1.21† 5.23±0.76 12.3±1.35‡
Heart (mg) 0.87±0.02 1.12±0.09* 0.71±0.02 0.82±0.03* 1.61±0.03 1.77±0.03* 1.19±0.03 1.56±0.10†
Liver (mg) 9.08±0.29 8.82±0.29 7.35±0.79 7.57±0.79 16.8±0.42 19.1±1.00 9.16±0.60 10.77±0.05
Table S1. Body weight, inguinal WAT, heart –, and liver weight in 5- and 12-month-old male and female rats treated with saline or leptin in neonatal period.
Body weight and organ weights 5 and 12 months
5 months 12 months
Weight male female male female
S-Tx L-Tx S-Tx L-Tx S-Tx L-Tx S-Tx L-Tx
Bodyweight
(g)
222.9±6.9 238.2±12.1 188.5±10.9 194.9±9.1 600.0±29.6 631.3±23.7 346.8±7.2 402.2±17.2†
Inguinal WAT
(mg)
1.32±0.15 2.03±0.35* 1.01±0.15 1.86±0.05† 5.31±0.41 9.25±1.21† 5.23±0.76 12.3±1.35‡
Heart (mg) 0.87±0.02 1.12±0.09* 0.71±0.02 0.82±0.03* 1.61±0.03 1.77±0.03* 1.19±0.03 1.56±0.10†
Liver (mg) 9.08±0.29 8.82±0.29 7.35±0.79 7.57±0.79 16.8±0.42 19.1±1.00 9.16±0.60 10.77±0.05
Table S1. Body weight, inguinal WAT, heart –, and liver weight in 5- and 12-month-old male and female rats treated with saline or leptin in neonatal period.
Sugar Preference Tests Performed at Day 30
Rats with experimental neonatal hyperleptinaemia have increased sucrose preference at Day 30
75
80
85
90
95
100
Males Females
% S
ucro
se P
refe
renc
e
Vehicle-treated
Leptin-treated*
Increased fat mass
Leptin
Dopamine -
Mesolimbic dopamine pathway
Lateral Hypothalamus
Neurotensin→ Orexin
Dopamine -
VTA Nucleus accumbens D2R D1R
Mesolimbic dopamine pathway
CB1 Mu Opioid
Decreased Palatable food consumption
Leptin’s Involvement in the Central Control of Mesolimbic Dopamine Pathway
Leinninger & Myers, 2008
0
5
10
15
20
25
30
35
NAcc Sh NAcc Core CPu Med CPu Lat Olf TubSpec
ific
[3H
]-Rac
lopr
ide
bind
ing
nCi/m
g
D2 Binding Males
Vehicle treated
Leptin-treated
*
0
5
10
15
20
25
30
35
NAcc Sh NAcc Core CPu Med CPu Lat Olf Tub
Spec
ific
[3H
]-Rac
lopr
ide
bind
ing
nCi/m
g
Brain Region
D2 Binding Females
* *
*
*
D2 receptor binding
Total binding: [3H]-Raclopride Non specific [3H]-Raclopride binding: +Butaclamol
0
2
4
6
8
10
12
NAcc Sh NAccCore
CPuPatches
CpuMatrix
PMCO VTA
Spec
fic [3 H
]-DAM
GO
b
indi
ng n
Ci/m
g
Mu Opioid Binding Males
Vehicle-treated
leptin-treated
*
0
2
4
6
8
10
12
NAcc Sh NAccCore
CPuPatches
CpuMatrix
PMCO VTA
Spec
fic [3 H
]-DAM
GO
b
indi
ng n
Ci/m
g
Mu Opioid Binding Females
Mu Opioid Receptor Binding
*Total Binding: [3H]-DAMGO *Non Specific [3H]-DAMGO Binding: +Naloxone
• Leptin receptors on dopaminergic neurons in the VTA and also on neurotensin neurons in the lateral hypothalamus influence dopamine VTA neurons (Leinninger & Myers, 2008).
• Leptin resistance at these sites may increase dopaminergic activity in the VTA, thereby leading to down-regulation of receptors in the nucleus accumbens.
• The extent to which these receptor changes underlie the increased sucrose preference remains to be determined.
Neonatal Hyperleptinaemia Influences Reward Pathways
• High leptin levels during a critical period for hypothalamic neurodevelopment can permanently influence the regulation of appetite and energy balance resulting in hyperphagia and obesity in the offspring of obese rodents.
• Experimental Hyperleptinaemia in the immediate postnatal period mimics the leptin resistant and obese phenotype we have previously described in offspring of obese rodents.
• Neonatal leptin exposure programmes a ‘selective leptin resistance’ in which the anorectic action of leptin is lost (the cardiovascular response to leptin is preserved).
• Hyperleptinaemia in the immediate postnatal period affects sucrose preference and reward-related receptor density in the mesolimbic dopamine pathway.
• Increased perinatal leptin exposure secondary to maternal obesity may predispose to increased appetite, altered food preference and risk of obesity.
Overall Summary 26
Maternal Obesity
Glucose, insulin, leptin, lipids, inflammatory response
increased neonatal adiposity Persistently altered energy
balance
Offspring Obesity
Transgenerational ‘Acceleration ‘ of Obesity?
Developmental origin of leptin resistance
Selective leptin resistance
↓ Metabolic action
↑ SNS activity
Hyperphagia & Obesity Hypertension
Excessive Weight Gain & Adiposity
Pregnancy & Lactation
Offspring Phenotype
Altered Leptin Signalling
Hypothalamus &
PVH NAc
Fetal/neonatal Hyperleptinaemia
& Hyperinsulinaemia?
Schematic: mechanisms in fetal programming of obesity and hypertension.
Relevance to human obesity ?
Rodents are altricial species: The relative maturity between the human brain and that of the rodent post-partum requires consideration.
Non-human primates and sheep support translation to precocial species and provide evidence for postnatal development of neurite projections.
Nothing is known about the sensitivity of the developing human hypothalamic circuitry to leptin in late pregnancy/early infancy.
Susceptibility to the neurotrophic influences of leptin could also occur in both ante-natal and post-partum periods.
Cord blood leptin concentration is elevated in neonates, falls rapidly post-partum, - developmental signal?.
Maternal obesity associated with raised cord blood leptin (Catalano et al 2009).
On-going RCT intervention studies in obese pregnant women offer a unique opportunity to study effects on childhood appetitive behaviour and obesity risk.
Thanks to:
Principle Investigators Professor Lucilla Poston (Tommy’s the Baby Charity) Professor Clive Coen (Professor of Neuroscience) Dr Paul Taylor (Senior Lecturer in Developmental Programming, HEFCE) Postdoctoral Research Assistants Dr Anne-Maj Samuelsson (Research Fellow, BHF) Dr Olena Rudyk (Research Fellow, BHF) Dr Shona Kirk (Research Fellow, BHF) Dr Tim South (BBSRC).
Research Assistant Mr Joaquim Plombo (British Heart Foundation) PhD Students Ms Phillippa Matthews (British Heart Foundation) Mr Marco Argenton (BHF) Ms Xanthi Maragkoudaki (BBRSC) Ms Angelina Mouralidarane Ms Abigail Morris (BBSRC) Collaborators Dr Nina Balthasar (UK), Dr Mike Christie (KCL) Dr Laura Cox (Texas) Dr Caroline Relton (UK) Dr Susan Ozanne (UK) Dr Eugene Jensen (BE) Dr Claude Remacle (BE), Dr Jude Oben (UCL).
Developmental Programming Research Group
Page 31
Page 32
Leptin impacts the peptidergic innervation of preautonomic divisions in adult Ob/Ob mouse PVH
Bouyer & Simerly J Neurosci. 2013 January 9; 33(2): 840–851.
Page 33
Maternal HFD Feeding Exclusively during Lactation Predisposes the Offspring for Metabolic Disorders
Vogt et al. Cell, Volume 156, Issue 3, 2014, 495 - 509
Maternal HFD Feeding Exclusively during Lactation Predisposes the Offspring for Metabolic Disorders and Impairs Axonal Projections of ARH Neurons to Intrahypothalamic Target Sites
Fiber Density PVHant
Fiber Density PVHpost
Vogt et al. Cell, Volume 156, Issue 3, 2014, 495 - 509
POMC-specific IR Deficiency in NCD/HFD offspring Rescues POMC Axonal Projections to Preautonomic Regions in the PVH
Milk Contents (Pre-weaning Stomach Contents) A
B
C
D
E
Lept
in (p
g/m
l)
0
50
100
150
200
250
2 7-8 9-11 13-14 15-18
**
****
#
#
#
FFA
(um
ol/g
)
0.0
100.0
200.0
300.0
400.0
500.0
600.0
2 7-8 9-11 13-14 15-18
*
**
†
†
# †
Cho
lest
erol
(um
ol/g
)
5.0
0.0
1.0
2.0
3.0
4.0
2 7-8 9-11 13-14 15-18
***
**
†
†
# Tr
igly
cerid
es (u
mol
/g)
0
50100
150
200250
300
2 7-8 9-11 13-14 15-18
**
**
**
†
†
Postnatal Day
Glu
cose
(um
ol/g
)
0.0
2.0
4.0
6.0
8.0
2 7-8 9-11 13-14 15-18
**
*
#
# †
A
B
C
D
E
Lept
in (p
g/m
l)
0
50
100
150
200
250
2 7-8 9-11 13-14 15-18
**
****
#
#
#
Lept
in (p
g/m
l)
0
50
100
150
200
250
2 7-8 9-11 13-14 15-18
**
****
0
50
100
150
200
250
2 7-8 9-11 13-14 15-18
**
****
#
#
#
FFA
(um
ol/g
)
0.0
100.0
200.0
300.0
400.0
500.0
600.0
2 7-8 9-11 13-14 15-18
*
**
†
†
# †
FFA
(um
ol/g
)
0.0
100.0
200.0
300.0
400.0
500.0
600.0
2 7-8 9-11 13-14 15-18
*
**
†
†
# †
Cho
lest
erol
(um
ol/g
)
5.0
0.0
1.0
2.0
3.0
4.0
2 7-8 9-11 13-14 15-18
***
**
†
†
# 0.0
1.0
2.0
3.0
4.0
2 7-8 9-11 13-14 15-18
***
**
†
†
# Tr
igly
cerid
es (u
mol
/g)
0
50100
150
200250
300
2 7-8 9-11 13-14 15-18
**
**
**
†
†
Trig
lyce
rides
(um
ol/g
)
0
50100
150
200250
300
2 7-8 9-11 13-14 15-18
**
**
**
†
†
Postnatal Day
Glu
cose
(um
ol/g
)
0.0
2.0
4.0
6.0
8.0
2 7-8 9-11 13-14 15-18
**
*
#
# †
Postnatal Day
Glu
cose
(um
ol/g
)
0.0
2.0
4.0
6.0
8.0
2 7-8 9-11 13-14 15-18
**
*
0.0
2.0
4.0
6.0
8.0
2 7-8 9-11 13-14 15-18
**
*
#
# †
Leptin
Cholesterol
Free Fatty Acids
Triglycerides
Glucose
Milk Contents (Pre-weaning Stomach Contents) A
B
C
D
E
Lept
in (p
g/m
l)
0
50
100
150
200
250
2 7-8 9-11 13-14 15-18
**
****
#
#
#
FFA
(um
ol/g
)
0.0
100.0
200.0
300.0
400.0
500.0
600.0
2 7-8 9-11 13-14 15-18
*
**
†
†
# †
Cho
lest
erol
(um
ol/g
)
5.0
0.0
1.0
2.0
3.0
4.0
2 7-8 9-11 13-14 15-18
***
**
†
†
# Tr
igly
cerid
es (u
mol
/g)
0
50100
150
200250
300
2 7-8 9-11 13-14 15-18
**
**
**
†
†
Postnatal Day
Glu
cose
(um
ol/g
)
0.0
2.0
4.0
6.0
8.0
2 7-8 9-11 13-14 15-18
**
*
#
# †
A
B
C
D
E
Lept
in (p
g/m
l)
0
50
100
150
200
250
2 7-8 9-11 13-14 15-18
**
****
#
#
#
Lept
in (p
g/m
l)
0
50
100
150
200
250
2 7-8 9-11 13-14 15-18
**
****
0
50
100
150
200
250
2 7-8 9-11 13-14 15-18
**
****
#
#
#
FFA
(um
ol/g
)
0.0
100.0
200.0
300.0
400.0
500.0
600.0
2 7-8 9-11 13-14 15-18
*
**
†
†
# †
FFA
(um
ol/g
)
0.0
100.0
200.0
300.0
400.0
500.0
600.0
2 7-8 9-11 13-14 15-18
*
**
†
†
# †
Cho
lest
erol
(um
ol/g
)
5.0
0.0
1.0
2.0
3.0
4.0
2 7-8 9-11 13-14 15-18
***
**
†
†
# 0.0
1.0
2.0
3.0
4.0
2 7-8 9-11 13-14 15-18
***
**
†
†
# Tr
igly
cerid
es (u
mol
/g)
0
50100
150
200250
300
2 7-8 9-11 13-14 15-18
**
**
**
†
†
Trig
lyce
rides
(um
ol/g
)
0
50100
150
200250
300
2 7-8 9-11 13-14 15-18
**
**
**
†
†
Postnatal Day
Glu
cose
(um
ol/g
)
0.0
2.0
4.0
6.0
8.0
2 7-8 9-11 13-14 15-18
**
*
#
# †
Postnatal Day
Glu
cose
(um
ol/g
)
0.0
2.0
4.0
6.0
8.0
2 7-8 9-11 13-14 15-18
**
*
0.0
2.0
4.0
6.0
8.0
2 7-8 9-11 13-14 15-18
**
*
#
# †
Leptin
Cholesterol
Free Fatty Acids
Triglycerides
Glucose
30 day Body weights and Organ weights in L-Tx rats 37
male female
S-Tx L-Tx S-Tx L-Tx
Body weight (g) 63.5±2.6 65.2±1.2 62.0±1.1 63.5±1.9
Inguinal WAT (mg) 0.18±0.03 0.20±0.03 0.15±0.03 0.14±0.01
Heart weight (mg) 0.35±0.02 0.41±0.01* 0.35±0.01 0.42±0.01*
Liver weight (mg) 2.53±0.13 2.58±0.10 2.33±0.10 2.60±0.32
Heart weight (mg/g BW) 5.53±0.17 6.34±0.28* 5.64±0.18 6.52±0.09*
Liver weight (mg/g BW) 40.2±2.9 39.1±1.7 38.7±2.0 42.3±2.6
1 Table 1. Body weight, inguinal WAT, and heart -weight in 30 day- old male and female rats treated with saline (S-Tx) or leptin (L-Tx) in neonatal period. Values given as mean+ SEM *P<0.05 versus saline using t-test, n=4-8.
30 day Body weights and Organ weights in L-Tx rats 38
male female
S-Tx L-Tx S-Tx L-Tx
Body weight (g) 63.5±2.6 65.2±1.2 62.0±1.1 63.5±1.9
Inguinal WAT (mg) 0.18±0.03 0.20±0.03 0.15±0.03 0.14±0.01
Heart weight (mg) 0.35±0.02 0.41±0.01* 0.35±0.01 0.42±0.01*
Liver weight (mg) 2.53±0.13 2.58±0.10 2.33±0.10 2.60±0.32
Heart weight (mg/g BW) 5.53±0.17 6.34±0.28* 5.64±0.18 6.52±0.09*
Liver weight (mg/g BW) 40.2±2.9 39.1±1.7 38.7±2.0 42.3±2.6
1 Table 1. Body weight, inguinal WAT, and heart -weight in 30 day- old male and female rats treated with saline (S-Tx) or leptin (L-Tx) in neonatal period. Values given as mean+ SEM *P<0.05 versus saline using t-test, n=4-8.
