SLIDE 1
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
Maternal Obesity and the Developmental Programming of Offspring - - PowerPoint PPT Presentation
Maternal Obesity and the Developmental Programming of Offspring - - PowerPoint PPT Presentation
Note: for non-commercial purposes only Womens Health Academic Centre 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.
SLIDE 2
SLIDE 3
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.
SLIDE 4
- Mice, rats, and humans share all but 1% of each
- ther'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
SLIDE 5
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
SLIDE 6
Maternal Characteristics
Page 5
B A
- 5-4-3-2-1
2 4 6 8 10 12 14 16 18 20
Body weight (g)
10 20 30 40 50
* **
Time (Gestational days) Time (weeks)
- 5-4-3-2-1
2 4 6 8 10 12 14 16 18 20
Body weight (g)
10 20 30 40 50
* **
Time (Gestational days) Time (weeks)
Calorific intake (kcal animal-1 day-1)
10 20 30 40
* *
- 5-4-3-2-1
2 4 6 8 10 12 14 16 18 20
Time (Gestational days) Time (weeks)
Calorific intake (kcal animal-1 day-1)
10 20 30 40
* *
- 5-4-3-2-1
2 4 6 8 10 12 14 16 18 20
Time (Gestational days) Time (weeks) Control Dam Obese Dam
E
9.8 3.6 Fat mass (g) 34.4 14.8 % Fat 18.8 20.5 Lean mass (g) OBESE CONTROL 9.8 3.6 Fat mass (g) 34.4 14.8 % Fat 18.8 20.5 Lean mass (g) OBESE CONTROL
C
BAT mass (g)
0.1 0.2 0.3
*
Control Obese
BAT mass (g)
0.1 0.2 0.3
*
Control Obese
WAT mass (g)
1 2 3 4 5 6 7 Control Obese
***
WAT mass (g)
1 2 3 4 5 6 7 Control Obese
WAT mass (g)
1 2 3 4 5 6 7 Control Obese
***
D
SLIDE 7
Maternal Characteristics
Page 6
Weaning
1 2 3 4 5 6
Triglyceride (mmol/L) Cholesterol (mmol/L) Glucose (10*mmol/L) Insulin (100*pmol/L) Leptin (100*pg/L)
Concentrations * ***
1 2 3 4 5 6
Triglyceride (mmol/L) Cholesterol (mmol/L) Glucose (10*mmol/L) Insulin (100*pmol/L) Leptin (100*pg/L)
Concentrations * ***
1 2 3 4 5 6
Triglyceride (mmol/L) Cholesterol (mmol/L) Glucose (10*mmol/L) Insulin (100*pmol/L) Leptin (10*pg/L)
Concentrations ** *** *** **
1 2 3 4 5 6
Triglyceride (mmol/L) Cholesterol (mmol/L) Glucose (10*mmol/L) Insulin (100*pmol/L) Leptin (10*pg/L)
Concentrations ** *** *** ** Late gestation
SLIDE 8
Offspring Phenotype: hyperphagia and adult obesity
Page 7
MALE FEMALE
Time (weeks)
5 10 15 20 25
body weight (g)
5 10 15 20 25 30 35
*
Time (weeks)
5 10 15 20 25
body weight (g)
5 10 15 20 25 30 35
*
5 10 15 20 25 5 10 15 20 25 30 35
*
Time (weeks)
5 10 15 20 25 5 10 15 20 25 30 35
*
Time (weeks) Time (weeks)
2 4 6 8 10 12 14 16 18 20 22
Calorific intake (kcal animal-1 week-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 mass (mg/g BW)
10 12 14 16 18 20 22
*** OC OO
Fat mass (mg/g BW)
6 8 10 12 14 16
**
1 2 3 4 5 6 Triglyceride (mmol/L) Cholesterol (mmol/L) Glucose (10*mmol/L) Insulin (100*pmol/L) Leptin (pg/L)
Concentrations
*** ** *
1 2 3 4 5 6 Triglyceride (mmol/L) Cholesterol (mmol/L) Glucose (10*mmol/L) Insulin (100*pmol/L) Leptin (pg/L)
Concentrations
* ** * * MALE FEMALE
Time (weeks)
5 10 15 20 25
body weight (g)
5 10 15 20 25 30 35
*
Time (weeks)
5 10 15 20 25
body weight (g)
5 10 15 20 25 30 35
*
5 10 15 20 25 5 10 15 20 25 30 35
*
Time (weeks)
5 10 15 20 25 5 10 15 20 25 30 35
*
Time (weeks) Time (weeks)
2 4 6 8 10 12 14 16 18 20 22
Calorific intake (kcal animal-1 week-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
Calorific intake (kcal animal-1 week-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 mass (mg/g BW)
10 12 14 16 18 20 22
*** OC OO
Fat mass (mg/g BW)
6 8 10 12 14 16
**
1 2 3 4 5 6 Triglyceride (mmol/L) Cholesterol (mmol/L) Glucose (10*mmol/L) Insulin (100*pmol/L) Leptin (pg/L)
Concentrations
*** ** *
1 2 3 4 5 6 Triglyceride (mmol/L) Cholesterol (mmol/L) Glucose (10*mmol/L) Insulin (100*pmol/L) Leptin (pg/L)
Concentrations
* ** * *
1 2 3 4 5 6 Triglyceride (mmol/L) Cholesterol (mmol/L) Glucose (10*mmol/L) Insulin (100*pmol/L) Leptin (pg/L)
Concentrations
*** ** *
1 2 3 4 5 6 Triglyceride (mmol/L) Cholesterol (mmol/L) Glucose (10*mmol/L) Insulin (100*pmol/L) Leptin (pg/L)
Concentrations
* ** * *
A B C D
SLIDE 9
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
SLIDE 10
Neonatal serum leptin and insulin in OffOb
Figure 1. Neonatal serum leptin and insulin concentrations in offspring
- f control and obese dams. Serum leptin (A) and insulin (B) were measured
in offspring of control dams (open bars) and obese dams (closed bars) over the suckling period. * p<0.05, ** p<0.01 and *** p<0.01 versus offspring of control dams for the same period (n = 3-6). For longitudinal comparisons, a significant difference (p<0.05) from the preceding period is indicated by # for
- ffspring of control dams and by † for offspring of obese dams.
1 2 3 4 5 6 7 2 7 8 9 11 13 14 15 Postnatal Day Serum Insulin (ng/ml) OffCon OffOb
** * * *** * * **
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 vs control, t-test, n=4-11.
1 2 3 4 5 6 7 2 7 8 9 11 13 14 15 Postnatal Day Serum Insulin (ng/ml) OffCon OffOb
** * * *** * * **
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 vs control, t-test, n=4-11. 2 7 8 9 11 13 14 15 18 10 20 30 40 50 Offspring of Obese Dams Offspring of Control Dams
*** *** *** *** *** *** *
Post natal Day Plasma Leptin Concentration (ng/ml) 2 7 8 9 11 13 14 15 18 10 20 30 40 50 Offspring of Obese Dams Offspring of Control Dams
*** *** *** *** *** *** *
Post natal Day Plasma Leptin Concentration (ng/ml)
A
Serum leptin (ng/ml)
*** *** *** *** *** *** *
B
1 2 3 4 5 6 7 2-8 9-11 13-18 Leptin mRNA expression (Copy # per geomean x 1000)
**
Postnatal day
† † † † # # # # # # †
2 7 8 9 11 13 14 15 18 10 20 30 40 50 Offspring of Obese Dams Offspring of Control Dams
*** *** *** *** *** *** *
Post natal Day Plasma Leptin Concentration (ng/ml) 2 7 8 9 11 13 14 15 18 10 20 30 40 50 Offspring of Obese Dams Offspring of Control Dams
*** *** *** *** *** *** *
Post natal Day Plasma Leptin Concentration (ng/ml)
A
Serum leptin (ng/ml)
*** *** *** *** *** *** *
A
Serum leptin (ng/ml) Serum leptin (ng/ml)
*** *** *** *** *** *** *
B
1 2 3 4 5 6 7 2-8 9-11 13-18 Leptin mRNA expression (Copy # per geomean x 1000)
**
Postnatal day
B
1 2 3 4 5 6 7 2-8 9-11 13-18 Leptin mRNA expression (Copy # per geomean x 1000)
**
Postnatal day 1 2 3 4 5 6 7 2-8 9-11 13-18 Leptin mRNA expression (Copy # per geomean x 1000)
**
Postnatal day
† † † † # # # # # # †
Postnatal Day Postnatal Day Serum Leptin (ng/ml) Serum Insulin (ng/ml) †
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 in
- ffspring from control (open bars) and obese dams (closed
bars). Day 2-8 (n=5-10), 9-11 (n=3-4), 13-18 (n=4-8) mRNA expressed as copy number divided by the geometric mean
- f 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 leptin mRNA expression (Copy #/geomean*1000) ** Post-natal day
SLIDE 11
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?
SLIDE 12
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
SLIDE 13
Leptin Challenge in Day 30 Offspring
12
5 10 15 20 25 30 35 40
OffCon
24 Hour Food Intake (g)
Leptin Saline
***
OffOb
5 10 15 20 25
OffOb
Change in Body Weight (g)
OffCon
*
Leptin Saline 5 10 15 20 25 30 35 40
OffOb
24 Hour Food Intake (g)
OffCon
**
Leptin Saline 2 4 6 8 10 12 14 16
OffOb OffCon
*
Change in Body Weight (g)
Leptin Saline
Males Females
24 hr Food Intake Δ Body Weight
Males Females
5 10 15 20 25 30 35 40
OffCon
24 Hour Food Intake (g)
Leptin Saline Leptin Saline
*** ***
OffOb
5 10 15 20 25
OffOb
Change in Body Weight (g)
OffCon
*
Leptin Saline Leptin Saline 5 10 15 20 25 30 35 40
OffOb
24 Hour Food Intake (g)
OffCon
** ** **
Leptin Saline Leptin Saline 2 4 6 8 10 12 14 16
OffOb OffCon
*
Change in Body Weight (g)
Leptin Saline Leptin Saline
Males Females
24 hr Food Intake Δ Body Weight
Males Females
SLIDE 14
A role for the melanocortin system in programming of appetite?
From Ozanne et al, Clinical Science 2005
SLIDE 15
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
OffOb OffCon ARC pSTAT3-ir Cells ~3.30 mm Caudal to Bregma ARC pSTAT3-ir Cells VMH pSTAT3-ir Cells
500 1000 1500 2000 2500 3000
- 2.30 -3.30
- 4.16
Number of pSTAT3-ir Cells/mm2
* 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
OffOb OffCon ARC pSTAT3-ir Cells ~3.30 mm Caudal to Bregma ARC pSTAT3-ir Cells VMH pSTAT3-ir Cells
500 1000 1500 2000 2500 3000
- 2.30 -3.30
- 4.16
Number of pSTAT3-ir Cells/mm2
* Bregma (mm)
SLIDE 16
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 AgRP-Immunoreactivity (% Area PVH)
OffOb OffCon AgRP Fluorescence (Bregma -1.80 mm) AgRP Fluorescence **
0.05 0.1 0.15 0.2 0.25 0.3
Bregma -1.80 mm AgRP-Immunoreactivity (% Area PVH)
**
0.05 0.1 0.15 0.2 0.25 0.3
Bregma -1.80 mm AgRP-Immunoreactivity (% Area PVH)
OffOb OffCon OffOb OffCon AgRP Fluorescence (Bregma -1.80 mm)
SLIDE 17
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.
SLIDE 18
Leptin Treatment in naïve rat pups Postnatal Day 9-15
Treatment PD9-15 PD2 PD7 PD8 PD9 PD11 PD20 0.0 0.1 0.2 0.3 Saline Leptin
** **
Postnatal Day Heart weight (g) Treatment PD9-15 PD2 PD7 PD8 PD9 PD11 PD12 PD14 PD20 20 40 60 80 100 Serum Leptin (ng/ml)
*** *** *** ***
leptin saline 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 offspring
- f control and obese dams. Serum leptin (A) and insulin (B) were measured
in offspring of control dams (open bars) and obese dams (closed bars) over the suckling period. * p<0.05, ** p<0.01 and *** p<0.01 versus offspring of control dams for the same period (n = 3-6). For longitudinal comparisons, a significant difference (p<0.05) from the preceding period is indicated by # for
- ffspring of control dams and by † for offspring of obese dams.
1 2 3 4 5 6 7 2 7 8 9 11 13 14 15 Postnatal Day Serum Insulin (ng/ml) OffCon OffOb
** * * *** * * **
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 vs control, t-test, n=4-11.
1 2 3 4 5 6 7 2 7 8 9 11 13 14 15 Postnatal Day Serum Insulin (ng/ml) OffCon OffOb
** * * *** * * **
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 vs control, t-test, n=4-11. 2 7 8 9 11 13 14 15 18 10 20 30 40 50 Offspring of Obese Dams Offspring of Control Dams
*** *** *** *** *** *** *
Post natal Day Plasma Leptin Concentration (ng/ml) 2 7 8 9 11 13 14 15 18 10 20 30 40 50 Offspring of Obese Dams Offspring of Control Dams
*** *** *** *** *** *** *
Post natal Day Plasma Leptin Concentration (ng/ml)
A
Serum leptin (ng/ml)
*** *** *** *** *** *** *
B
1 2 3 4 5 6 7 2-8 9-11 13-18 Leptin mRNA expression (Copy # per geomean x 1000)
**
Postnatal day
† † † † # # # # # # †
2 7 8 9 11 13 14 15 18 10 20 30 40 50 Offspring of Obese Dams Offspring of Control Dams
*** *** *** *** *** *** *
Post natal Day Plasma Leptin Concentration (ng/ml) 2 7 8 9 11 13 14 15 18 10 20 30 40 50 Offspring of Obese Dams Offspring of Control Dams
*** *** *** *** *** *** *
Post natal Day Plasma Leptin Concentration (ng/ml)
A
Serum leptin (ng/ml)
*** *** *** *** *** *** *
A
Serum leptin (ng/ml) Serum leptin (ng/ml)
*** *** *** *** *** *** *
B
1 2 3 4 5 6 7 2-8 9-11 13-18 Leptin mRNA expression (Copy # per geomean x 1000)
**
Postnatal day
B
1 2 3 4 5 6 7 2-8 9-11 13-18 Leptin mRNA expression (Copy # per geomean x 1000)
**
Postnatal day 1 2 3 4 5 6 7 2-8 9-11 13-18 Leptin mRNA expression (Copy # per geomean x 1000)
**
Postnatal day
† † † † # # # # # # †
Postnatal Day Postnatal Day Serum Leptin (ng/ml) Serum Insulin (ng/ml) †
1 2 3 4 5 6 2 7 8 9 11 13 14 15
A B
SLIDE 19
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
- f
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
*
5 10 15 20 Food intake (g) S-Tx L-Tx
** *
5 10 15 Weight gain (g) S-Tx L-Tx
Saline injection Leptin bolus injection
Male
S-Lx L-Tx 5 10 15 20 25 Weight gain (g)
*
Female Male Female Male
**
S-Tx L-Tx 10 20 30 40 Food intake (g)
Female Male
S-Tx L-Tx 5 10 15 Weight gain (g)
**
5 10 15 20 Food intake (g) S-Tx L-Tx
*
Female
5 10 15 20 25 Weight gain (g) S-Tx L-Tx S-Tx L-Tx 10 20 30 40 Food intake (g)
** *
Leptin Challenge in L-Tx neonates at 30 days and 5 months
SLIDE 20
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.
SLIDE 21
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.
SLIDE 22
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 % Sucrose Preference Vehicle-treated Leptin-treated *
SLIDE 23
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
SLIDE 24
5 10 15 20 25 30 35 NAcc Sh NAcc Core CPu Med CPu Lat Olf Tub Specific [3H]-Raclopride binding nCi/mg
D2 Binding Males
Vehicle treated Leptin-treated
*
5 10 15 20 25 30 35 NAcc Sh NAcc Core CPu Med CPu Lat Olf Tub Specific [3H]-Raclopride binding nCi/mg Brain Region
D2 Binding Females
* * * *
D2 receptor binding
Total binding: [3H]-Raclopride Non specific [3H]-Raclopride binding: +Butaclamol
SLIDE 25
2 4 6 8 10 12 NAcc Sh NAcc Core CPu Patches Cpu Matrix PMCO VTA
Specfic [3H]-DAMGO binding nCi/mg
Mu Opioid Binding Males
Vehicle-treated leptin-treated
*
2 4 6 8 10 12 NAcc Sh NAcc Core CPu Patches Cpu Matrix PMCO VTA
Specfic [3H]-DAMGO binding nCi/mg
Mu Opioid Binding Females
Mu Opioid Receptor Binding
*Total Binding: [3H]-DAMGO *Non Specific [3H]-DAMGO Binding: +Naloxone
SLIDE 26
- Leptin receptors on dopaminergic neurons in the VTA and also
- n 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
SLIDE 27
- 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
- ffspring 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
- besity.
Overall Summary
26
SLIDE 28
Maternal Obesity
Glucose, insulin, leptin, lipids, inflammatory response increased neonatal adiposity Persistently altered energy balance
Offspring Obesity Transgenerational ‘Acceleration ‘ of Obesity?
SLIDE 29
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.
SLIDE 30
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
- ccur 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.
SLIDE 31
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
SLIDE 32
Page 31
SLIDE 33
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.
SLIDE 34
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
SLIDE 35
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
SLIDE 36
Milk Contents (Pre-weaning Stomach Contents)
A B C D E
Leptin (pg/ml) 50 100 150 200 250 2 7-8 9-11 13-14 15-18
**
** **
# # #
FFA (umol/g) 0.0 100.0 200.0 300.0 400.0 500.0 600.0 2 7-8 9-11 13-14 15-18
* **
† † # †
Cholesterol (umol/g) 5.0 0.0 1.0 2.0 3.0 4.0 2 7-8 9-11 13-14 15-18
* ** **
† † #
Triglycerides (umol/g) 50 100 150 200 250 300 2 7-8 9-11 13-14 15-18
** ** * *
† † Postnatal Day
Glucose (umol/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
Leptin (pg/ml) 50 100 150 200 250 2 7-8 9-11 13-14 15-18
**
** **
# # #
Leptin (pg/ml) 50 100 150 200 250 2 7-8 9-11 13-14 15-18
**
** **
50 100 150 200 250 2 7-8 9-11 13-14 15-18
**
** **
# # #
FFA (umol/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 (umol/g) 0.0 100.0 200.0 300.0 400.0 500.0 600.0 2 7-8 9-11 13-14 15-18
* **
† † # †
Cholesterol (umol/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
* ** **
† † #
Triglycerides (umol/g) 50 100 150 200 250 300 2 7-8 9-11 13-14 15-18
** ** * *
† †
Triglycerides (umol/g) 50 100 150 200 250 300 2 7-8 9-11 13-14 15-18
** ** * *
† † Postnatal Day
Glucose (umol/g) 0.0 2.0 4.0 6.0 8.0 2 7-8 9-11 13-14 15-18
** *
# # † Postnatal Day
Glucose (umol/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
SLIDE 37
Milk Contents (Pre-weaning Stomach Contents)
A B C D E
Leptin (pg/ml) 50 100 150 200 250 2 7-8 9-11 13-14 15-18
**
** **
# # #
FFA (umol/g) 0.0 100.0 200.0 300.0 400.0 500.0 600.0 2 7-8 9-11 13-14 15-18
* **
† † # †
Cholesterol (umol/g) 5.0 0.0 1.0 2.0 3.0 4.0 2 7-8 9-11 13-14 15-18
* ** **
† † #
Triglycerides (umol/g) 50 100 150 200 250 300 2 7-8 9-11 13-14 15-18
** ** * *
† † Postnatal Day
Glucose (umol/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
Leptin (pg/ml) 50 100 150 200 250 2 7-8 9-11 13-14 15-18
**
** **
# # #
Leptin (pg/ml) 50 100 150 200 250 2 7-8 9-11 13-14 15-18
**
** **
50 100 150 200 250 2 7-8 9-11 13-14 15-18
**
** **
# # #
FFA (umol/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 (umol/g) 0.0 100.0 200.0 300.0 400.0 500.0 600.0 2 7-8 9-11 13-14 15-18
* **
† † # †
Cholesterol (umol/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
* ** **
† † #
Triglycerides (umol/g) 50 100 150 200 250 300 2 7-8 9-11 13-14 15-18
** ** * *
† †
Triglycerides (umol/g) 50 100 150 200 250 300 2 7-8 9-11 13-14 15-18
** ** * *
† † Postnatal Day
Glucose (umol/g) 0.0 2.0 4.0 6.0 8.0 2 7-8 9-11 13-14 15-18
** *
# # † Postnatal Day
Glucose (umol/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
SLIDE 38
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.
SLIDE 39
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