Biological Determinants:

Developmental Origins

Peter Nathanielsz, Peter Nathanielsz,

Center for Pregnancy and Center for Pregnancy and

Newborn Research,Newborn Research,

University of Texas Health University of Texas Health

Sciences Center.Sciences Center.

San AntonioSan Antonio

Biological Determinants:

Developmental Origins

1. Overview of developmental programming and evidence for the importance of the developmental environment in determining offspring – particularly female offspring phenotype.

2. Mechanisms whereby a female offspring’s phenotype is changed by her developmental environment thus putting her own later pregnancy at risk.

3. Transgenerational effects.

4. Action.

ProgrammingThe process through which a stimulus or

insult establishes a persistent response.

Developmental programming

hypothesisExposure during a critical period in

development may influence phenotype -

life time function and health.

Developmental ProgrammingDevelopmental Programming

The idea that our lifeThe idea that our life--time health is as much time health is as much

impacted by the conditions under which we impacted by the conditions under which we

develop in the womb and early life as by our develop in the womb and early life as by our

genes is a paradigm shift in our thinking of genes is a paradigm shift in our thinking of

what needs to be done to improve the health of what needs to be done to improve the health of

the nation. We need to think in terms of genethe nation. We need to think in terms of gene--

environment interactions. environment interactions.

We pass more biological milestones

before we are born and in the early

months as a baby than at any other

time in our lives.

How we pass these milestones will

be greatly affected by our very first

environments, the womb and

immediately after birth.

Biological Determinants:

Developmental Origins

1. Overview of developmental programming and evidence Overview of developmental programming and evidence for the importance of the developmental environment for the importance of the developmental environment in determining offspring in determining offspring –– particularly female offspring particularly female offspring phenotype.phenotype.

2. Mechanisms whereby a female offspring’s phenotype is changed by her developmental environment thus putting her own later pregnancy at risk.

3. Transgenerational effects.

4. Action.

Blood cholesterol in men and women at the age

of 50 can be related to the extent to which they

had to protect their brain when they were fetuses

How do we know that

nutrition in pregnancy

is important?

A

young

Pima

woman

High Type 2

Diabetes

Low Type 2

Diabetes

Biological Determinants:

Developmental Origins1. Overview of developmental programming and evidence

for the importance of the developmental environment in determining offspring – particularly female offspring phenotype.

2. Mechanisms whereby offspring’s phenotype is changed 2. Mechanisms whereby offspring’s phenotype is changed by her developmental environment thus putting her by her developmental environment thus putting her own later pregnancy at risk.own later pregnancy at risk.

3. Transgenerational effects.

4. Action.

The couch potato rat.The couch potato rat.

Severe maternal undernutrition (30% normal diet) induces

increased fat mass and reduces offspring locomotor activity. Vickers MH et al., 2000;2001; 2003

Voluntary locomotor activity

Cardiovascular variables allometry and plasma

analytes in six month old offspring of fat mice.Samuelsson, A.-M. et al. Hypertension 2008;51:383-392

Immunohistochemistry in 0.9 G fetal baboon

pancreas CTR (A-C) and MNR (D-F). IGF-1 (A,D);

IGF-2 (B,E) and insulin (C,F).

A B C

D E F

0

0.5

1

1.5

2

2.5

3

CTR MNR

% o

f are

a s

tain

ed

0

4000000

8000000

12000000

16000000

CTR MNRD

en

sit

y

Fetal pancreatic Insulin fraction and density from ad lib fed controls baboons (CTR, n=7) or nutrient

restricted (MNR; fed 70% CTR; n=7 ) from 0.16 to 0.9G. Data M ± SEM. *p< 0.05.

Insulin Protein in 0.9G Baboon Fetal Pancreas

*

*

Effects of maternal obesity on fetal Effects of maternal obesity on fetal

development in the sheep.development in the sheep.

The sheep is widely used as a

biomedical model for human

pregnancy, and is the only model

large enough to study the challenged fetus directly combined

with an evaluation of post-natal effects.

Methods

• Multiparous Western Whiteface ewes carrying singleton fetuses were fed a high energy diet at 100% (controls) or 150% (Obese) of NRC recommendations from 60 days before conception to day 75 of gestation.

• A subset of ewes were subjected to a GTT on day 75.

• A second set of ewes were necropsied on day 75 of gestation and the fetus recovered.

• A third set of ewes were allowed to deliver.

The first half of gestation is critical for:

1) Placental growth and vascularization

2) Fetal organogenesis: fetal organs shown

to be most affected by early gestational under nutrition include the pancreas.

0.00%

1.00%

2.00%

3.00%

4.00%

5.00%

6.00%

Cot Car

Va

sc

ula

r a

rea

de

ns

ity

Con OB

*

*

Vascularity in both CAR and COT tissues of OB and C ewes at d 75 of gestation

Relative Content of FATP1,FATP4 and CD36 protein in COT of Relative Content of FATP1,FATP4 and CD36 protein in COT of

Control and OB Ewes on Day 75 of GestationControl and OB Ewes on Day 75 of Gestation

FATP1

FATP4

Tubulin

C C OB OB

CD36

:control fed ewes; obese over-nourished ewes:

Mean ± SEM; **: p<0.01; n=5 in each group.

Placental macrophages in obese women. A: placental sections from a lean (pre-gravid BMI: 22.5) woman

-rare Macrophage staining; an obese (pre-gravid BMI: 31.7) woman with increased numbers of

macrophages (C) and (D). Right : Placental macrophage number. M ± SE. *: p < 0.001 obese vs lean.

Lower panel: macrophages (thick arrows) in the villous stroma Thin arrow: syncytiotrophoblast nuclei,

#: fetal blood space, *: maternal blood space. Scale 20 µm. J.C. Challier et al Placenta 29 274-281

Concentration of Glucose and Insulin in

Maternal Blood on 0.5 Gestation in the obese

sheep model.

0

50

100

150

200

250

-20 0 20 40 60 80 100 120 140

Time (min)

glu

co

se (

mg

/dL

)

OB

C

Area Under the Curve differs by dietary treatment (P<0.05)

0.00

0.50

1.00

1.50

2.00

2.50

3.00

-20 0 20 40 60 80 100 120 140

Time (min)

Insu

lin

(n

g/m

L)

OB

C

C OB C OB0

10

20

30

40

50

60

70

Maternal Fetal

*

*

Glu

co

se (

mg

/dl)

Glucose Content in Both Maternal and Fetal Blood

on D75 of Gestation

*Means ± SEM differ (P<0.05)

Fetal beta-cells in

- obese mother

Fetal beta-cells in

control mother

Beta cells in the pancreas

of the fetal sheep

Insulin Content in Both Maternal and Fetal Blood

on D75 of Gestation

*Means ± SEM differ (P<0.05)

C OB C OB0.0

0.5

1.0

1.5

Maternal Fetal

*

*Insu

lin

(n

g/m

L)

Con fetal muscle OB fetal muscle

Effect of obesogenic diet on fetal muscle Effect of obesogenic diet on fetal muscle

histologyhistology

AA B

Cort

isol (n

g/m

l)

0

5

10

15

20

Cort

isol (n

g/m

l)

0

5

10

15

20

CTR NR Fat CTR NR Fat

*

Cortisol in Control (CTR, n=6) maternal nutrient restricted (NR, n=6)and overfed (Fat, N=6) ewes and fetuses at 78 days of gestation.

* p< 0.05 compared to CTR group.

*

*

MATERNAL FETAL

Fetal plasma cortisol at 0.5 G

* *

Post natal data

We have evaluated glucose We have evaluated glucose

handling at two months of age.handling at two months of age.

Concentration of Glucose and

Insulin in Lambs at 2 Months of Age

Time (min)

BL 2 5 10 15 30 60

Glu

co

se

(m

g/d

L)

0

50

100

150

200

250

Compared to Control group (C): *P<0.05; + p=0.07.

0

50

100

150

COB

C OB

AU

C (

mg/d

l/h)

*

Time (min)

BL 2 5 10 15 30 60

Insulin

(µ U

/mL)

0

10

20

30

0

10

20

30

40

COB

*

AU

C (

mg/d

l/h)

C OB

+

Gillman MW et al, JAMA 2001 285, 2461-2467

Risk of Overweight in Adolescence By Duration of

Breastfeeding in Infancy

What are the possible

mechanisms

for the programming of a

predisposition to obesity?

Yura et al Cell

Metab. 2005

Jun;1(6):371-8.

Effects of maternal dietary protein restriction and folic acid

supplementation during pregnancy in rats on gene promoter methylation

and RNA expression in the offspring 34 d after birth

Diets: C 18% protein, 1mg/kg folic acid ( ); R – 9% protein, 1 mg/kg folic acid ( );

RF – 9% protein, 5mg/kg folic acid ( ). Values are mean ± SD, n=10 offspring/

maternal dietary group. P< 0.05: a) vs C; b) vs RF.

0

20

40

60

80

100

120

% C

ontr

ol

0

200

400

1000

1200

1400

GR PPARα PPARγ GR PPARα PPARγ ACOX

ab ab

ab

ab

ab

DNA methylation mRNA concentration

Lillycrop K et al.

Biological Determinants:

Developmental Origins1. Overview of developmental programming and evidence for

the importance of the developmental environment in determining offspring – particularly female offspring phenotype.

2. Mechanisms whereby a female offspring’s phenotype is changed by her developmental environment thus putting her own later pregnancy at risk.

3. 3. TransgenerationalTransgenerational effects.effects.

4. Action.

Windows of Development

CC RR CR RC

At birth litters were adjusted to 12 pups/litter

Zambrano et al. (2005) Journal of Physiology 566: 225-236

GrowthGestation Lactationday0 22 0 21 ~130

Control ControlCC

RR Restricted

CR

RC

~270

Time line

~90

Control

Control

Control

Restricted

Restricted

Restricted

Control

Control

Control

Puberty

Mothers Pups

Adult life

Programming of Obesity

Zambrano et al. (2005) Journal of Physiology 566: 225-236

CC

RC

CC RC

22 month old female offspring from dams fed control (CC) and

restricted (RC) diets

IRI

0

2

4

6

8

a,b

a

b

CC RR CR RC

a*

*

IRI

0

5

10

15

20

CC RR CR RC

aa

a

b

*

*

Insulin Resistance Index (IRI) in grand daughters and

grandsons of rats fed either control (C - 20%)protein or restricted (R – 10%) protein diet during

pregnancy (first letter) or lactation (second letter); mean + SEM; n=12 -16.

Granddaughters Grandsons

Transgenerational effects of maternal under nutritionon insulin resistance in granddaughters and grandsons.

Biological Determinants:

Developmental Origins1. Overview of developmental programming and evidence for

the importance of the developmental environment in determining offspring – particularly female offspring phenotype.

2. Mechanisms whereby a female offspring’s phenotype is changed by her developmental environment thus putting her own later pregnancy at risk.

3. Transgenerational effects.

4. Action.

ACKNOWLEDGEMENTSACKNOWLEDGEMENTSTo all members of the Center for Pregnancy and To all members of the Center for Pregnancy and

Newborn Research, University of Texas Health Newborn Research, University of Texas Health

Science Center, San Antonio.Science Center, San Antonio.

• Steve Ford

• Elena Zambrano

• Natalia Schlabritz

• Cun Li

• Thomas McDonald