kerstik. linask, ph.d. university of south florida ... · university of south florida children’s...
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Alcohol -Induced Birth Defects and Folate Supplementation in
Mouse Model
KERSTI K. LINASK, Ph.D.
University of South Florida
Children’s Research Institute
Fetal Alcohol Spectrum Disorders (FASD) is a serious health and social
problem which affects individuals, families, and societies worldwide.
FASD is characterized by language and motor delays, cognitive delays
including intellectual disabilities, facial abnormalities, heart defects, and
vision and hearing problems
FASD afflicts millions of people worldwide – about one in every
100 births (May et al., 2009)
FASD Occurs World-wide …
� FASD can include Fetal Alcohol Syndrome
(FAS), Fetal Alcohol Effects, Alcohol-Related
Neurodevelopmental Disorder (ARND) and
Central Nervous System (CNS) problems.
� Fewer than 10% of FAS individuals are able
to take care of themselves and live on their
own. The annual total cost to society for the
care of children and adults with Fetal Alcohol
Syndrome is huge: in the US it is between 1.4
billion and 9.7 billion.
Statistics on Drinking…
Are alcohol dependent (alcoholics)
Are dependent in that they put themselves or
others at risk: binge drinking
women 4 drinks/occasion
men 5 drinks/occasion
On average 8 drinks are reported/occasion and
4 times per month
Those who drink moderately or do not drink
WHO: number 37 with 8.77 liters of pure alcohol consumption per capita every year
Statistics on Drinking - Eestis: Based on World Health Organization Global
Information on Alcohol and Health. Shown is alcohol per capita consumption based in
liters of pure alcohol drank per year between 1990-2010
The top 15 countries of alcohol consumption:
1. Belarus 14.37
2. Andorra 13.31
3. Lithuania 12.9
4. Czech Republic 12.69
5. Grenada 12.4
6. Austria 12.1
7. Ireland 11.92
8. France 11.7
9. Saint Lucia 11.6
10. Estonia 11.36
11. Luxembourg 11.36
12. Germany 11.21
13. Russian Federation 11.12
14. Slovakia 10.96
15. Portugal 10.84
Beer 4.68 liters (41.2%)
Wine 1.26 l (11.1%)
Spirits 4.18 l (36.8%
Other 1.24 l (10.9 %)
What is a Standard Drink?
From: Langman’s Essential Medical Embryology by T.W.Sadler, Lippincott,
Williams, and Wilkins, 2005
3 Weeks
Moore and Persaud. 2003
16 days Human Embryo
(from Visual embryo
website)
21 days
5 to 6 weeks:
pregnancy is usually
confirmed.
During Pregnancy ….
ChickFish Human
Vertebrate Embryos at Early Stages are Similar
Critical Periods in Human Prenatal Period
Gastrulation: IngressionHuman 17 days Mouse (Mu) 6.75 – 7.5 days
Hu 18 days Mu 7.50 – 8.0 days
Hu 20-21 days Mu 8-8.5 days
EP
MES
ECT
ME
S
EN
D
Neural Development
During gastrulation,
signaling, including
Wnt/beta-catenin, induces
the neural compartment in
the ectoderm layer
overlying the mesoderm
layer and this induction
occurs in an
anterior/posterior
progression initiating cell
specification and cell
differentiation down the
neural tube.Stiles and Jernigan, 2010
Congenital Heart Disease
� Congenital heart malformations, the most common of human birth defects, occur in ~ 1% of the population worldwide.
� Valve and associated structures continue to be the most common subtype of CHDs accounting for 25-30% of all cardiovascular malformations.
� Cardiac anomalies appear to arise from abnormal
� cell lineage decisions of early progenitor cells, disrupting
� normal patterning of morphogenetic pathways.
A recent recommendation suggested that children born with
cardiac defects also be tested for neural-related problems and
developmental disabilities (Marino et al., 2012).
This underscores that common developmental processes are
involved in both cardiac and neural development occurring at
the same time of development during gastrulation.
Alcohol (ethanol) consumption during pregnancy is
linked to congenital heart defects and to neural-related
problems that are associated with Fetal Alcohol
Syndrome (FAS).
Gastrulation: E13 to E20
Cardiac progenitor cells Neural progenitor cells
Multiple axis, no cardiac tissueHeart anterior to head
Cardiabifida
Looping defects, wide hearts
Adverse effects are stage- and time- dependent
11-11.5 hrs / Early St.3
11.5 – 12 hrs / Mid St. 3
12 – 12.5 hrs / Late St. 3
to St. 4
Total embryos analyzed: 451
CONTROL
11 hrs after fertilization
(~ HH early St-3)
11.5 hrs after fertilization
(~ HH mid St-3)
By the end of the embryonic period,
week 8, the rudimentary structures
of the brain and central nervous
system are established and the
major compartments of the central
and peripheral nervous systems are
defined.
- Primitive patterning of
sensorimotor regions of the
neural compartment within the
neocortex is established (Bishop et
al., 2002);
- major compartments within
diencephalic and midbrain have
differentiated (Nakamura et al., 2005)
and &
- segmental organization of
hindbrain and spinal column are
specified (Gavalos et al., 2003).
Neural Development
Acute Ethanol Exposure on Day 9: Effects on Mouse Brain Development
High resolution
magnetic resonance
imaging study:
From: Parnell et al., 2013 Neurotox. Teratol. 39:77-83
Exposure: On day 9 (Hu D20)
of mouse gestation, pregnant
mouse given two i.p. injections
4 hrs apart of 2.9 g/kg/dose
ethanol (25% vol/vol) or of
Ringer’s solution.
Color-coding for significance:• yellow=slightly different p<0.05
• orange = more changes, p<0.01
• red = changes highly significant
p<0.005
Epidemiological studies suggest
that 54% of live-born children with
FAS have some kind of cardiac
anomaly. The malformations
include valvuloseptal defects,
stenosis of the pulmonary artery,
Tetralogy of Fallot, and d-
transposition of the great arteries.
Gastrulation: IngressionHuman 17 days Mouse (Mu) 6.75 – 7.5 days
Hu 18 days Mu 7.50 – 8.0 days
Hu 20-21 days Mu 8-8.5 days
EP
MES
ECT
MES
END
Neural Development
During gastrulation, signaling,
including Wnt/beta-catenin,
induces the neural
compartment in the ectoderm
layer overlying the mesoderm
layer and this induction occurs
in an anterior/posterior
progression initiating cell
specification and cell
differentiation down the neural
tube.
Stiles and Jernigan, 2010
Congenital Heart Disease
• Congenital heart malformations, the most common of human birth defects, occur in ~ 1% of the population worldwide.
• Valve and associated structures continue to be the most common subtype of CHDs accounting for 25-30% of all cardiovascular malformations.
• Cardiac anomalies appear to arise from abnormal
• cell lineage decisions of early progenitor cells, disrupting
• normal patterning of morphogenetic pathways.
A recent recommendation suggested that children born with cardiac
defects also be tested for neural-related problems and developmental
disabilities (Marino et al., 2012).
This underscores that common developmental processes are involved in
both cardiac and neural development occurring at the same time of
development during gastrulation.
Alcohol (ethanol) consumption during pregnancy is linked to
congenital heart defects and to neural-related problems that
are associated with Fetal Alcohol Syndrome (FAS).
Gastrulation: E13 to E20
Cardiac progenitor cells Neural progenitor cells
Multiple axis, no cardiac tissueHeart anterior to head
Cardiabifida
Looping defects, wide hearts
Adverse effects are stage- and time- dependent
11-11.5 hrs / Early St.3
11.5 – 12 hrs / Mid St. 3
12 – 12.5 hrs / Late St. 3
to St. 4
Total embryos analyzed: 451
CONTROL
11 hrs after fertilization
(~ HH early St-3)
11.5 hrs after fertilization
(~ HH mid St-3)
By the end of the embryonic period,
week 8, the rudimentary structures of
the brain and central nervous system
are established and the major
compartments of the central and
peripheral nervous systems are
defined.
- Primitive patterning of sensorimotor
regions of the neural compartment
within the neocortex is established
(Bishop et al., 2002);
- major compartments within
diencephalic and midbrain have
differentiated (Nakamura et al., 2005)
and …
- segmental organization of hindbrain
and spinal column are specified (Gavalos et al., 2003).
Neural Development
Acute Ethanol Exposure on Day 9: Effects on Mouse Brain Development
High resolution magnetic
resonance imaging study:
From: Parnell et al., 2013 Neurotox. Teratol. 39:77-83
Exposure: On day 9 (Hu D20) of
mouse gestation, pregnant mouse
given two i.p. injections 4 hrs
apart of 2.9 g/kg/dose ethanol
(25% vol/vol) or of Ringer’s
solution.
Color-coding for significance:• yellow=slightly different p<0.05
• orange = more changes, p<0.01
• red = changes highly significant
p<0.005
Epidemiological studies suggest that
54% of live-born children with FAS have
some kind of cardiac anomaly. The
malformations include valvuloseptal
defects, stenosis of the pulmonary
artery, Tetralogy of Fallot, and d-
transposition of the great arteries.
Moore and Persaud. 2003
16 days Human Embryo
(from Visual embryo
website)3 Weeks
49% of pregnancies are unintended! Walker, D., et al., Fetal Alcohol Spectrum Disorders Prevention: An exploratory
study of women's use of, attitudes toward, and knowledge about alcohol. J Amer Acad Nurse Practitioners, 2005. 17(5): p. 187-193.
Gastrulation (between embryonic days E 13 to E 20) should be considered a high risk
period of pregnancy, as a woman most likely is not yet unaware of her pregnancy and
is not taking precautionary measures to reduce exposure risks to the embryo, as for
example, by not taking certain drugs, not drinking alcohol, and not smoking.
The mother also may not have started folic acid dietary supplementation known to
decrease risk of birth defects.
Early cardiac
progenitor
cells begin to
differentiate in
the heart
fields.
21 days
During gastrulation, Wnt/beta-
catenin signaling and the
induction of the heart
compartment occur in an
anterior/posterior
progression initiating cardiac
cell specification and cell
sorting across the
bilateral regions
of the ventral mesoderm layer.
Linask et al., 1992; 1997;
2003
Heart Organogenesis
Environmental exposures
induce similar cardiac defects
in vertebrate animal models as
in the human depending upon
developmental timing of
exposure and dose.
Defining the effects of alcohol on heart development
in the mouse model.
• Pregnant mice were exposed
acutely to an accepted binge
drinking level of EtOH (two i.p. injections of 2.9 g EtOH/kg maternal
weight, given 4 hrs apart).
• Exposure targeted ED 6.75
(gastrulation).
METHODOLOGY:
ED6.75
ED15.5
On ED 15.5 of gestation doppler echocardiography was used to
determine effects of alcohol on embryonic mouse heart
function in control and ethanol exposed embryos.
A binge drinking exposure of ethanol on ED 6.75 resulted in
abnormal fetal cardiac function and valve defects.
Arrhythmia Atrioventricular Valve Regurgitation
Semilunar Valve Regurgitation Semilunar Valve Stenosis
Blue signal is active Wnt/β-catenin signaling using the TopGal transgenic line.
Question: Does folic acid when provided early in
gestation on morning after conception reverse the
adverse effects of EtOH on mouse heart
development,?
Serrano, M., Brinez, P., Han, M., Lastra-Vicente, R., and K.K. Linask. 2010. Fetal Alcohol
Syndrome: Cardiac birth defects and prevention with folate. Amer. J. Obst. Gyn. 203(1):75.e7-
75.e15.
Folic Acid Supplementation
Mouse Model: Dietary folate supplementation (10
mg/kg or 6.2 mg/kg) was started on morning of
vaginal plug detection.
Only the high dose of folate provided complete protection
of cardiac defects.
A combination of folate and myo-inositol is more effective
in embryonic protection than folate alone.
Microarray Analysis of Cardiac Gene Expression after Hcys or Li Environmental Exposure and Folate Protection:
46,000 genes analyzed- partial list shown
Total RNA extracted
cDNA
Hybridization to Affymetrix
Gene Chips
Right ventricle
and
outflow tract
isolated
Gender-Associated Changes: Gene Ontology of Specific Biological Processes
Fatty acid β-oxidation
Organic acid catabolic proc
Carboxylic acid catab proc
ATP biosynthesis
Fatty acid catab processes
Lipid oxidation
Fatty acid metabolic proc
Coenzyme metabolic proc
Energy coupled proton trans
Electron transport chain
Cofactor metabolic proc
The bioinformatic
analyses indicated
lipid metabolism is
most affected and the
misexpression can
have a gender bias for
specific genes.
Table 2. Gender -Associated Changes: Gene Ontology (GO) Specific Biological Processes
GO ID Description hcys_f1_
vs_cont_f
1
hcys_f1_
vs_fol_f1
lith_f1_vs
_cont_f1
lith_f1_vs
_fol_f1
lith_f1_vs
_lith_fol_
f1
lith_f1_vs
_lith_fol_
f2
fol_f1_vs_
cont_f1
hcys_m1
_vs_cont
_m1
hcys_m1
_vs_fol_
m1
hcys_m1
_vs_hcys
_fol_m1
hcys_m1
_vs_hcys
_fol_m2
lith_m1_
vs_cont_
m1
lith_m1_
vs_fol_m
1
fol_m1_vs
_cont_m1
Number
of
instances
GO:0007186 G-protein coupled receptor signaling pathwayNS INCONSISTENTUP UP NS NS NS UP UP NS UP UP UP DOWN 9
GO:0006754 ATP biosynthetic process UP UP NS UP NS NS DOWN NS DOWN DOWN DOWN NS NS UP 8
GO:0003012 muscle system process UP UP UP UP NS NS DOWN NS NS NS DOWN NS NS UP 7
GO:0006635 fatty acid beta-oxidation NS NS NS DOWN NS NS NS DOWN DOWN DOWN DOWN DOWN DOWN NS 7
GO:0007155 cell adhesion NS DOWN NS NS DOWN NS NS UP NS UP UP NS DOWN UP 7
GO:0007218 neuropeptide signaling pathway UP DOWN UP NS NS NS UP NS NS NS UP NS UP DOWN 7
GO:0009201 ribonucleoside triphosphate biosynthetic processUP UP NS NS NS NS DOWN NS DOWN DOWN DOWN NS NS UP 7
GO:0009206 purine ribonucleoside triphosphate biosynthetic processUP UP NS NS NS NS DOWN NS DOWN DOWN DOWN NS NS UP 7
GO:0015985 energy coupled proton transport, down electrochemical gradientUP UP NS UP NS NS DOWN NS NS DOWN DOWN NS NS UP 7
GO:0015986 ATP synthesis coupled proton transportUP UP NS UP NS NS DOWN NS NS DOWN DOWN NS NS UP 7
GO:0016054 organic acid catabolic process NS NS NS DOWN NS NS NS DOWN DOWN DOWN DOWN DOWN DOWN NS 7
GO:0022610 biological adhesion NS DOWN NS NS DOWN NS NS UP NS UP UP NS DOWN UP 7
GO:0046395 carboxylic acid catabolic process NS NS NS DOWN NS NS NS DOWN DOWN DOWN DOWN DOWN DOWN NS 7
GO:0055114 oxidation-reduction process UP UP NS NS NS NS NS DOWN DOWN DOWN DOWN NS DOWN NS 7
GO:0071804 cellular potassium ion transport NS UP UP UP NS NS DOWN UP UP NS NS UP NS NS 7
GO:0071805 potassium ion transmembrane transportNS UP UP UP NS NS DOWN UP UP NS NS UP NS NS 7
GO:0006082 organic acid metabolic process NS NS NS DOWN NS NS NS DOWN DOWN DOWN DOWN NS DOWN NS 6
GO:0006412 translation UP NS NS NS NS NS NS NS DOWN DOWN DOWN NS DOWN UP 6
GO:0006457 protein folding NS NS NS NS NS NS NS DOWN DOWN DOWN DOWN DOWN DOWN NS 6
GO:0006818 hydrogen transport UP UP NS UP NS NS DOWN NS NS DOWN DOWN NS NS NS 6
GO:0006936 muscle contraction UP UP NS UP NS NS DOWN NS NS NS DOWN NS NS UP 6
GO:0009062 fatty acid catabolic process NS NS NS NS NS NS NS DOWN DOWN DOWN DOWN DOWN DOWN NS 6
GO:0009145 purine nucleoside triphosphate biosynthetic processUP UP NS NS NS NS NS NS DOWN DOWN DOWN NS NS UP 6
GO:0019395 fatty acid oxidation NS NS NS NS NS NS NS DOWN DOWN DOWN DOWN DOWN DOWN NS 6
GO:0019752 carboxylic acid metabolic process NS NS NS DOWN NS NS NS DOWN DOWN DOWN DOWN NS DOWN NS 6
GO:0021545 cranial nerve development NS DOWN NS NS NS DOWN DOWN UP NS UP UP NS NS NS 6
GO:0030198 extracellular matrix organization NS DOWN UP NS NS NS UP NS NS UP UP NS NS UP 6
GO:0032787 monocarboxylic acid metabolic processNS NS NS DOWN NS NS NS DOWN DOWN DOWN DOWN NS DOWN NS 6
GO:0034440 lipid oxidation NS NS NS NS NS NS NS DOWN DOWN DOWN DOWN DOWN DOWN NS 6
GO:0043062 extracellular structure organization NS DOWN UP NS NS NS UP NS NS UP UP NS NS UP 6
GO:0043436 oxoacid metabolic process NS NS NS DOWN NS NS NS DOWN DOWN DOWN DOWN NS DOWN NS 6
GO:0044281 small molecule metabolic process NS NS NS DOWN NS NS NS DOWN DOWN DOWN DOWN NS DOWN NS 6
GO:0044282 small molecule catabolic process NS NS NS DOWN NS NS NS DOWN DOWN DOWN DOWN NS DOWN NS 6
GO:0044712 single-organism catabolic process NS NS NS DOWN NS NS NS DOWN DOWN DOWN DOWN NS DOWN NS 6
GO:0060349 bone morphogenesis NS DOWN UP NS NS NS UP UP UP NS UP NS NS NS 6
GO:0061053 somite development NS DOWN NS NS NS NS NS UP UP UP UP NS UP NS 6
GO:0072329 monocarboxylic acid catabolic processNS NS NS NS NS NS NS DOWN DOWN DOWN DOWN DOWN DOWN NS 6
GO:0001973 adenosine receptor signaling pathwayNS NS NS NS NS NS DOWN UP UP NS NS UP UP NS 5
GO:0006091 generation of precursor metabolites and energyUP UP NS NS NS NS NS NS DOWN DOWN DOWN NS NS NS 5
GO:0006310 DNA recombination NS NS NS NS NS NS NS DOWN DOWN DOWN DOWN DOWN NS NS 5
GO:0006631 fatty acid metabolic process NS NS NS DOWN NS NS NS DOWN DOWN DOWN DOWN NS NS NS 5
GO:0006732 coenzyme metabolic process NS NS DOWN NS NS NS NS DOWN DOWN DOWN DOWN NS NS NS 5
GO:0006941 striated muscle contraction UP UP NS UP NS NS DOWN NS NS NS NS NS NS UP 5
GO:0009108 coenzyme biosynthetic process DOWN NS DOWN DOWN DOWN NS NS NS NS NS DOWN NS NS NS 5
GO:0009142 nucleoside triphosphate biosynthetic processUP UP NS NS NS NS NS NS DOWN DOWN DOWN NS NS NS 5
GO:0015672 monovalent inorganic cation transportUP UP NS UP NS NS DOWN NS NS NS NS NS NS UP 5
GO:0022900 electron transport chain UP UP NS NS NS NS NS NS DOWN DOWN DOWN NS NS NS 5
GO:0030317 sperm motility UP UP NS NS DOWN DOWN DOWN NS NS NS NS NS NS NS 5
GO:0033539 fatty acid beta-oxidation using acyl-CoA dehydrogenaseNS NS NS NS NS NS NS DOWN DOWN DOWN DOWN DOWN NS NS 5
GO:0035282 segmentation NS DOWN NS NS NS NS NS UP UP UP UP NS NS NS 5
GO:0044710 single-organism metabolic process NS NS NS DOWN NS NS NS NS DOWN DOWN DOWN NS DOWN NS 5
GO:0051186 cofactor metabolic process NS NS NS DOWN NS NS NS DOWN DOWN DOWN DOWN NS NS NS 5
GO:0055085 transmembrane transport UP UP NS UP NS NS DOWN NS NS NS NS UP NS NS 5
GO ID Description hcys_f1_
vs_cont_f
1
hcys_f1_
vs_fol_f1
lith_f1_vs
_cont_f1
lith_f1_vs
_fol_f1
lith_f1_vs
_lith_fol_
f1
lith_f1_vs
_lith_fol_
f2
fol_f1_vs_
cont_f1
hcys_m1
_vs_cont
_m1
hcys_m1
_vs_fol_
m1
hcys_m1
_vs_hcys
_fol_m1
hcys_m1
_vs_hcys
_fol_m2
lith_m1_
vs_cont_
m1
lith_m1_
vs_fol_m
1
fol_m1_vs
_cont_m1
Number
of
instances
GO:0007186 G-protein coupled receptor signaling pathwayNS INCONSISTENTUP UP NS NS NS UP UP NS UP UP UP DOWN 9
GO:0006754 ATP biosynthetic process UP UP NS UP NS NS DOWN NS DOWN DOWN DOWN NS NS UP 8
GO:0003012 muscle system process UP UP UP UP NS NS DOWN NS NS NS DOWN NS NS UP 7
GO:0006635 fatty acid beta-oxidation NS NS NS DOWN NS NS NS DOWN DOWN DOWN DOWN DOWN DOWN NS 7
GO:0007155 cell adhesion NS DOWN NS NS DOWN NS NS UP NS UP UP NS DOWN UP 7
GO:0007218 neuropeptide signaling pathway UP DOWN UP NS NS NS UP NS NS NS UP NS UP DOWN 7
GO:0009201 ribonucleoside triphosphate biosynthetic processUP UP NS NS NS NS DOWN NS DOWN DOWN DOWN NS NS UP 7
GO:0009206 purine ribonucleoside triphosphate biosynthetic processUP UP NS NS NS NS DOWN NS DOWN DOWN DOWN NS NS UP 7
GO:0015985 energy coupled proton transport, down electrochemical gradientUP UP NS UP NS NS DOWN NS NS DOWN DOWN NS NS UP 7
GO:0015986 ATP synthesis coupled proton transportUP UP NS UP NS NS DOWN NS NS DOWN DOWN NS NS UP 7
GO:0016054 organic acid catabolic process NS NS NS DOWN NS NS NS DOWN DOWN DOWN DOWN DOWN DOWN NS 7
GO:0022610 biological adhesion NS DOWN NS NS DOWN NS NS UP NS UP UP NS DOWN UP 7
GO:0046395 carboxylic acid catabolic process NS NS NS DOWN NS NS NS DOWN DOWN DOWN DOWN DOWN DOWN NS 7
GO:0055114 oxidation-reduction process UP UP NS NS NS NS NS DOWN DOWN DOWN DOWN NS DOWN NS 7
GO:0071804 cellular potassium ion transport NS UP UP UP NS NS DOWN UP UP NS NS UP NS NS 7
GO:0071805 potassium ion transmembrane transportNS UP UP UP NS NS DOWN UP UP NS NS UP NS NS 7
GO:0006082 organic acid metabolic process NS NS NS DOWN NS NS NS DOWN DOWN DOWN DOWN NS DOWN NS 6
GO:0006412 translation UP NS NS NS NS NS NS NS DOWN DOWN DOWN NS DOWN UP 6
GO:0006457 protein folding NS NS NS NS NS NS NS DOWN DOWN DOWN DOWN DOWN DOWN NS 6
GO:0006818 hydrogen transport UP UP NS UP NS NS DOWN NS NS DOWN DOWN NS NS NS 6
GO:0006936 muscle contraction UP UP NS UP NS NS DOWN NS NS NS DOWN NS NS UP 6
GO:0009062 fatty acid catabolic process NS NS NS NS NS NS NS DOWN DOWN DOWN DOWN DOWN DOWN NS 6
GO:0009145 purine nucleoside triphosphate biosynthetic processUP UP NS NS NS NS NS NS DOWN DOWN DOWN NS NS UP 6
GO:0019395 fatty acid oxidation NS NS NS NS NS NS NS DOWN DOWN DOWN DOWN DOWN DOWN NS 6
GO:0019752 carboxylic acid metabolic process NS NS NS DOWN NS NS NS DOWN DOWN DOWN DOWN NS DOWN NS 6
GO:0021545 cranial nerve development NS DOWN NS NS NS DOWN DOWN UP NS UP UP NS NS NS 6
GO:0030198 extracellular matrix organization NS DOWN UP NS NS NS UP NS NS UP UP NS NS UP 6
GO:0032787 monocarboxylic acid metabolic processNS NS NS DOWN NS NS NS DOWN DOWN DOWN DOWN NS DOWN NS 6
GO:0034440 lipid oxidation NS NS NS NS NS NS NS DOWN DOWN DOWN DOWN DOWN DOWN NS 6
GO:0043062 extracellular structure organization NS DOWN UP NS NS NS UP NS NS UP UP NS NS UP 6
GO:0043436 oxoacid metabolic process NS NS NS DOWN NS NS NS DOWN DOWN DOWN DOWN NS DOWN NS 6
GO:0044281 small molecule metabolic process NS NS NS DOWN NS NS NS DOWN DOWN DOWN DOWN NS DOWN NS 6
GO:0044282 small molecule catabolic process NS NS NS DOWN NS NS NS DOWN DOWN DOWN DOWN NS DOWN NS 6
GO:0044712 single-organism catabolic process NS NS NS DOWN NS NS NS DOWN DOWN DOWN DOWN NS DOWN NS 6
GO:0060349 bone morphogenesis NS DOWN UP NS NS NS UP UP UP NS UP NS NS NS 6
GO:0061053 somite development NS DOWN NS NS NS NS NS UP UP UP UP NS UP NS 6
GO:0072329 monocarboxylic acid catabolic processNS NS NS NS NS NS NS DOWN DOWN DOWN DOWN DOWN DOWN NS 6
GO:0001973 adenosine receptor signaling pathwayNS NS NS NS NS NS DOWN UP UP NS NS UP UP NS 5
GO:0006091 generation of precursor metabolites and energyUP UP NS NS NS NS NS NS DOWN DOWN DOWN NS NS NS 5
GO:0006310 DNA recombination NS NS NS NS NS NS NS DOWN DOWN DOWN DOWN DOWN NS NS 5
GO:0006631 fatty acid metabolic process NS NS NS DOWN NS NS NS DOWN DOWN DOWN DOWN NS NS NS 5
GO:0006732 coenzyme metabolic process NS NS DOWN NS NS NS NS DOWN DOWN DOWN DOWN NS NS NS 5
GO:0006941 striated muscle contraction UP UP NS UP NS NS DOWN NS NS NS NS NS NS UP 5
GO:0009108 coenzyme biosynthetic process DOWN NS DOWN DOWN DOWN NS NS NS NS NS DOWN NS NS NS 5
GO:0009142 nucleoside triphosphate biosynthetic processUP UP NS NS NS NS NS NS DOWN DOWN DOWN NS NS NS 5
GO:0015672 monovalent inorganic cation transportUP UP NS UP NS NS DOWN NS NS NS NS NS NS UP 5
GO:0022900 electron transport chain UP UP NS NS NS NS NS NS DOWN DOWN DOWN NS NS NS 5
GO:0030317 sperm motility UP UP NS NS DOWN DOWN DOWN NS NS NS NS NS NS NS 5
GO:0033539 fatty acid beta-oxidation using acyl-CoA dehydrogenaseNS NS NS NS NS NS NS DOWN DOWN DOWN DOWN DOWN NS NS 5
GO:0035282 segmentation NS DOWN NS NS NS NS NS UP UP UP UP NS NS NS 5
GO:0044710 single-organism metabolic process NS NS NS DOWN NS NS NS NS DOWN DOWN DOWN NS DOWN NS 5
GO:0051186 cofactor metabolic process NS NS NS DOWN NS NS NS DOWN DOWN DOWN DOWN NS NS NS 5
GO:0055085 transmembrane transport UP UP NS UP NS NS DOWN NS NS NS NS UP NS NS 5
Moore and Persaud. 2003
16 days Human Embryo
(from Visual embryo
website)3 Weeks
49% of pregnancies are unintended! Walker, D., et al., Fetal Alcohol Spectrum Disorders Prevention:
An exploratory study of women's use of, attitudes toward, and knowledge about alcohol. J Amer Acad Nurse Practitioners, 2005. 17(5): p. 187-193.
Gastrulation (between embryonic days E 13 to E 20) should be considered
a high risk period of pregnancy, as a woman most likely is not yet unaware
of her pregnancy and is not taking precautionary measures to reduce
exposure risks to the embryo, as for example, by not taking certain drugs,
not drinking alcohol, and not smoking.
The mother also may not have started folic acid dietary supplementation
known to decrease risk of birth defects.
Early cardiac
progenitor
cells begin to
differentiate in
the heart
fields.
21 days
During gastrulation,
Wnt/beta-catenin signaling
and the induction of the
heart compartment occur in
an anterior/posterior
progression initiating
cardiac cell specification
and cell sorting across the
bilateral regions
of the ventral mesoderm
layer.
Linask et al., 1992; 1997;
2003
Heart Organogenesis
Environmental exposures
induce similar cardiac defects
in vertebrate animal models as
in the human depending upon
developmental timing of
exposure and dose.
Defining the effects of alcohol on heart development
in the mouse model.
• Pregnant mice were exposed
acutely to an accepted binge
drinking level of EtOH (two i.p. injections of 2.9 g EtOH/kg maternal
weight, given 4 hrs apart).
• Exposure targeted ED 6.75
(gastrulation).
METHODOLOGY:
ED6.75
ED15.5
On ED 15.5 of gestation doppler echocardiography
was used to determine effects of alcohol on
embryonic mouse heart function in control and
ethanol exposed embryos.
A binge drinking exposure of ethanol on ED 6.75
resulted in abnormal fetal cardiac function and valve
defects.
Arrhythmia Atrioventricular Valve Regurgitation
Semilunar Valve Regurgitation Semilunar Valve Stenosis
Blue signal is active Wnt/β-catenin signaling using the TopGal transgenic line.
Question: Does folic acid when provided
early in gestation on morning after conception
reverse the adverse effects of EtOH on
mouse heart development,?
Serrano, M., Brinez, P., Han, M., Lastra-Vicente, R., and K.K. Linask. 2010. Fetal
Alcohol Syndrome: Cardiac birth defects and prevention with folate. Amer. J. Obst.
Gyn. 203(1):75.e7-75.e15.
Folic Acid Supplementation
Mouse Model: Dietary folate supplementation
(10 mg/kg or 6.2 mg/kg) was started on
morning of vaginal plug detection.
Only the high dose of folate provided complete
protection of cardiac defects.
A combination of folate and myo-inositol is more
effective in embryonic protection than folate
alone.
Microarray Analysis of Cardiac Gene Expression after Hcys or Li Environmental Exposure and Folate Protection:
46,000 genes analyzed- partial list shown
Total RNA extracted
cDNA
Hybridization to Affymetrix
Gene Chips
Right
ventricle and
outflow tract
isolated
Gender-Associated Changes: Gene Ontology of Specific Biological Processes
Fatty acid β-oxidation
Organic acid catabolic proc
Carboxylic acid catab proc
ATP biosynthesis
Fatty acid catab processes
Lipid oxidation
Fatty acid metabolic proc
Coenzyme metabolic proc
Energy coupled proton trans
Electron transport chain
Cofactor metabolic proc
The bioinformatic
analyses indicated
lipid metabolism is
most affected and
the misexpression
can have a gender
bias for specific
genes.
Table 2. Gender -Associated Changes: Gene Ontology (GO) Specific Biological Processes
GO ID Description hcys_f1_
vs_cont_f
1
hcys_f1_
vs_fol_f1
lith_f1_vs
_cont_f1
lith_f1_vs
_fol_f1
lith_f1_vs
_lith_fol_
f1
lith_f1_vs
_lith_fol_
f2
fol_f1_vs_
cont_f1
hcys_m1
_vs_cont
_m1
hcys_m1
_vs_fol_
m1
hcys_m1
_vs_hcys
_fol_m1
hcys_m1
_vs_hcys
_fol_m2
lith_m1_
vs_cont_
m1
lith_m1_
vs_fol_m
1
fol_m1_vs
_cont_m1
Number
of
instances
GO:0007186 G-protein coupled receptor signaling pathwayNS INCONSISTENTUP UP NS NS NS UP UP NS UP UP UP DOWN 9
GO:0006754 ATP biosynthetic process UP UP NS UP NS NS DOWN NS DOWN DOWN DOWN NS NS UP 8
GO:0003012 muscle system process UP UP UP UP NS NS DOWN NS NS NS DOWN NS NS UP 7
GO:0006635 fatty acid beta-oxidation NS NS NS DOWN NS NS NS DOWN DOWN DOWN DOWN DOWN DOWN NS 7
GO:0007155 cell adhesion NS DOWN NS NS DOWN NS NS UP NS UP UP NS DOWN UP 7
GO:0007218 neuropeptide signaling pathway UP DOWN UP NS NS NS UP NS NS NS UP NS UP DOWN 7
GO:0009201 ribonucleoside triphosphate biosynthetic processUP UP NS NS NS NS DOWN NS DOWN DOWN DOWN NS NS UP 7
GO:0009206 purine ribonucleoside triphosphate biosynthetic processUP UP NS NS NS NS DOWN NS DOWN DOWN DOWN NS NS UP 7
GO:0015985 energy coupled proton transport, down electrochemical gradientUP UP NS UP NS NS DOWN NS NS DOWN DOWN NS NS UP 7
GO:0015986 ATP synthesis coupled proton transportUP UP NS UP NS NS DOWN NS NS DOWN DOWN NS NS UP 7
GO:0016054 organic acid catabolic process NS NS NS DOWN NS NS NS DOWN DOWN DOWN DOWN DOWN DOWN NS 7
GO:0022610 biological adhesion NS DOWN NS NS DOWN NS NS UP NS UP UP NS DOWN UP 7
GO:0046395 carboxylic acid catabolic process NS NS NS DOWN NS NS NS DOWN DOWN DOWN DOWN DOWN DOWN NS 7
GO:0055114 oxidation-reduction process UP UP NS NS NS NS NS DOWN DOWN DOWN DOWN NS DOWN NS 7
GO:0071804 cellular potassium ion transport NS UP UP UP NS NS DOWN UP UP NS NS UP NS NS 7
GO:0071805 potassium ion transmembrane transportNS UP UP UP NS NS DOWN UP UP NS NS UP NS NS 7
GO:0006082 organic acid metabolic process NS NS NS DOWN NS NS NS DOWN DOWN DOWN DOWN NS DOWN NS 6
GO:0006412 translation UP NS NS NS NS NS NS NS DOWN DOWN DOWN NS DOWN UP 6
GO:0006457 protein folding NS NS NS NS NS NS NS DOWN DOWN DOWN DOWN DOWN DOWN NS 6
GO:0006818 hydrogen transport UP UP NS UP NS NS DOWN NS NS DOWN DOWN NS NS NS 6
GO:0006936 muscle contraction UP UP NS UP NS NS DOWN NS NS NS DOWN NS NS UP 6
GO:0009062 fatty acid catabolic process NS NS NS NS NS NS NS DOWN DOWN DOWN DOWN DOWN DOWN NS 6
GO:0009145 purine nucleoside triphosphate biosynthetic processUP UP NS NS NS NS NS NS DOWN DOWN DOWN NS NS UP 6
GO:0019395 fatty acid oxidation NS NS NS NS NS NS NS DOWN DOWN DOWN DOWN DOWN DOWN NS 6
GO:0019752 carboxylic acid metabolic process NS NS NS DOWN NS NS NS DOWN DOWN DOWN DOWN NS DOWN NS 6
GO:0021545 cranial nerve development NS DOWN NS NS NS DOWN DOWN UP NS UP UP NS NS NS 6
GO:0030198 extracellular matrix organization NS DOWN UP NS NS NS UP NS NS UP UP NS NS UP 6
GO:0032787 monocarboxylic acid metabolic processNS NS NS DOWN NS NS NS DOWN DOWN DOWN DOWN NS DOWN NS 6
GO:0034440 lipid oxidation NS NS NS NS NS NS NS DOWN DOWN DOWN DOWN DOWN DOWN NS 6
GO:0043062 extracellular structure organization NS DOWN UP NS NS NS UP NS NS UP UP NS NS UP 6
GO:0043436 oxoacid metabolic process NS NS NS DOWN NS NS NS DOWN DOWN DOWN DOWN NS DOWN NS 6
GO:0044281 small molecule metabolic process NS NS NS DOWN NS NS NS DOWN DOWN DOWN DOWN NS DOWN NS 6
GO:0044282 small molecule catabolic process NS NS NS DOWN NS NS NS DOWN DOWN DOWN DOWN NS DOWN NS 6
GO:0044712 single-organism catabolic process NS NS NS DOWN NS NS NS DOWN DOWN DOWN DOWN NS DOWN NS 6
GO:0060349 bone morphogenesis NS DOWN UP NS NS NS UP UP UP NS UP NS NS NS 6
GO:0061053 somite development NS DOWN NS NS NS NS NS UP UP UP UP NS UP NS 6
GO:0072329 monocarboxylic acid catabolic processNS NS NS NS NS NS NS DOWN DOWN DOWN DOWN DOWN DOWN NS 6
GO:0001973 adenosine receptor signaling pathwayNS NS NS NS NS NS DOWN UP UP NS NS UP UP NS 5
GO:0006091 generation of precursor metabolites and energyUP UP NS NS NS NS NS NS DOWN DOWN DOWN NS NS NS 5
GO:0006310 DNA recombination NS NS NS NS NS NS NS DOWN DOWN DOWN DOWN DOWN NS NS 5
GO:0006631 fatty acid metabolic process NS NS NS DOWN NS NS NS DOWN DOWN DOWN DOWN NS NS NS 5
GO:0006732 coenzyme metabolic process NS NS DOWN NS NS NS NS DOWN DOWN DOWN DOWN NS NS NS 5
GO:0006941 striated muscle contraction UP UP NS UP NS NS DOWN NS NS NS NS NS NS UP 5
GO:0009108 coenzyme biosynthetic process DOWN NS DOWN DOWN DOWN NS NS NS NS NS DOWN NS NS NS 5
GO:0009142 nucleoside triphosphate biosynthetic processUP UP NS NS NS NS NS NS DOWN DOWN DOWN NS NS NS 5
GO:0015672 monovalent inorganic cation transportUP UP NS UP NS NS DOWN NS NS NS NS NS NS UP 5
GO:0022900 electron transport chain UP UP NS NS NS NS NS NS DOWN DOWN DOWN NS NS NS 5
GO:0030317 sperm motility UP UP NS NS DOWN DOWN DOWN NS NS NS NS NS NS NS 5
GO:0033539 fatty acid beta-oxidation using acyl-CoA dehydrogenaseNS NS NS NS NS NS NS DOWN DOWN DOWN DOWN DOWN NS NS 5
GO:0035282 segmentation NS DOWN NS NS NS NS NS UP UP UP UP NS NS NS 5
GO:0044710 single-organism metabolic process NS NS NS DOWN NS NS NS NS DOWN DOWN DOWN NS DOWN NS 5
GO:0051186 cofactor metabolic process NS NS NS DOWN NS NS NS DOWN DOWN DOWN DOWN NS NS NS 5
GO:0055085 transmembrane transport UP UP NS UP NS NS DOWN NS NS NS NS UP NS NS 5
GO ID Description hcys_f1_
vs_cont_f
1
hcys_f1_
vs_fol_f1
lith_f1_vs
_cont_f1
lith_f1_vs
_fol_f1
lith_f1_vs
_lith_fol_
f1
lith_f1_vs
_lith_fol_
f2
fol_f1_vs_
cont_f1
hcys_m1
_vs_cont
_m1
hcys_m1
_vs_fol_
m1
hcys_m1
_vs_hcys
_fol_m1
hcys_m1
_vs_hcys
_fol_m2
lith_m1_
vs_cont_
m1
lith_m1_
vs_fol_m
1
fol_m1_vs
_cont_m1
Number
of
instances
GO:0007186 G-protein coupled receptor signaling pathwayNS INCONSISTENTUP UP NS NS NS UP UP NS UP UP UP DOWN 9
GO:0006754 ATP biosynthetic process UP UP NS UP NS NS DOWN NS DOWN DOWN DOWN NS NS UP 8
GO:0003012 muscle system process UP UP UP UP NS NS DOWN NS NS NS DOWN NS NS UP 7
GO:0006635 fatty acid beta-oxidation NS NS NS DOWN NS NS NS DOWN DOWN DOWN DOWN DOWN DOWN NS 7
GO:0007155 cell adhesion NS DOWN NS NS DOWN NS NS UP NS UP UP NS DOWN UP 7
GO:0007218 neuropeptide signaling pathway UP DOWN UP NS NS NS UP NS NS NS UP NS UP DOWN 7
GO:0009201 ribonucleoside triphosphate biosynthetic processUP UP NS NS NS NS DOWN NS DOWN DOWN DOWN NS NS UP 7
GO:0009206 purine ribonucleoside triphosphate biosynthetic processUP UP NS NS NS NS DOWN NS DOWN DOWN DOWN NS NS UP 7
GO:0015985 energy coupled proton transport, down electrochemical gradientUP UP NS UP NS NS DOWN NS NS DOWN DOWN NS NS UP 7
GO:0015986 ATP synthesis coupled proton transportUP UP NS UP NS NS DOWN NS NS DOWN DOWN NS NS UP 7
GO:0016054 organic acid catabolic process NS NS NS DOWN NS NS NS DOWN DOWN DOWN DOWN DOWN DOWN NS 7
GO:0022610 biological adhesion NS DOWN NS NS DOWN NS NS UP NS UP UP NS DOWN UP 7
GO:0046395 carboxylic acid catabolic process NS NS NS DOWN NS NS NS DOWN DOWN DOWN DOWN DOWN DOWN NS 7
GO:0055114 oxidation-reduction process UP UP NS NS NS NS NS DOWN DOWN DOWN DOWN NS DOWN NS 7
GO:0071804 cellular potassium ion transport NS UP UP UP NS NS DOWN UP UP NS NS UP NS NS 7
GO:0071805 potassium ion transmembrane transportNS UP UP UP NS NS DOWN UP UP NS NS UP NS NS 7
GO:0006082 organic acid metabolic process NS NS NS DOWN NS NS NS DOWN DOWN DOWN DOWN NS DOWN NS 6
GO:0006412 translation UP NS NS NS NS NS NS NS DOWN DOWN DOWN NS DOWN UP 6
GO:0006457 protein folding NS NS NS NS NS NS NS DOWN DOWN DOWN DOWN DOWN DOWN NS 6
GO:0006818 hydrogen transport UP UP NS UP NS NS DOWN NS NS DOWN DOWN NS NS NS 6
GO:0006936 muscle contraction UP UP NS UP NS NS DOWN NS NS NS DOWN NS NS UP 6
GO:0009062 fatty acid catabolic process NS NS NS NS NS NS NS DOWN DOWN DOWN DOWN DOWN DOWN NS 6
GO:0009145 purine nucleoside triphosphate biosynthetic processUP UP NS NS NS NS NS NS DOWN DOWN DOWN NS NS UP 6
GO:0019395 fatty acid oxidation NS NS NS NS NS NS NS DOWN DOWN DOWN DOWN DOWN DOWN NS 6
GO:0019752 carboxylic acid metabolic process NS NS NS DOWN NS NS NS DOWN DOWN DOWN DOWN NS DOWN NS 6
GO:0021545 cranial nerve development NS DOWN NS NS NS DOWN DOWN UP NS UP UP NS NS NS 6
GO:0030198 extracellular matrix organization NS DOWN UP NS NS NS UP NS NS UP UP NS NS UP 6
GO:0032787 monocarboxylic acid metabolic processNS NS NS DOWN NS NS NS DOWN DOWN DOWN DOWN NS DOWN NS 6
GO:0034440 lipid oxidation NS NS NS NS NS NS NS DOWN DOWN DOWN DOWN DOWN DOWN NS 6
GO:0043062 extracellular structure organization NS DOWN UP NS NS NS UP NS NS UP UP NS NS UP 6
GO:0043436 oxoacid metabolic process NS NS NS DOWN NS NS NS DOWN DOWN DOWN DOWN NS DOWN NS 6
GO:0044281 small molecule metabolic process NS NS NS DOWN NS NS NS DOWN DOWN DOWN DOWN NS DOWN NS 6
GO:0044282 small molecule catabolic process NS NS NS DOWN NS NS NS DOWN DOWN DOWN DOWN NS DOWN NS 6
GO:0044712 single-organism catabolic process NS NS NS DOWN NS NS NS DOWN DOWN DOWN DOWN NS DOWN NS 6
GO:0060349 bone morphogenesis NS DOWN UP NS NS NS UP UP UP NS UP NS NS NS 6
GO:0061053 somite development NS DOWN NS NS NS NS NS UP UP UP UP NS UP NS 6
GO:0072329 monocarboxylic acid catabolic processNS NS NS NS NS NS NS DOWN DOWN DOWN DOWN DOWN DOWN NS 6
GO:0001973 adenosine receptor signaling pathwayNS NS NS NS NS NS DOWN UP UP NS NS UP UP NS 5
GO:0006091 generation of precursor metabolites and energyUP UP NS NS NS NS NS NS DOWN DOWN DOWN NS NS NS 5
GO:0006310 DNA recombination NS NS NS NS NS NS NS DOWN DOWN DOWN DOWN DOWN NS NS 5
GO:0006631 fatty acid metabolic process NS NS NS DOWN NS NS NS DOWN DOWN DOWN DOWN NS NS NS 5
GO:0006732 coenzyme metabolic process NS NS DOWN NS NS NS NS DOWN DOWN DOWN DOWN NS NS NS 5
GO:0006941 striated muscle contraction UP UP NS UP NS NS DOWN NS NS NS NS NS NS UP 5
GO:0009108 coenzyme biosynthetic process DOWN NS DOWN DOWN DOWN NS NS NS NS NS DOWN NS NS NS 5
GO:0009142 nucleoside triphosphate biosynthetic processUP UP NS NS NS NS NS NS DOWN DOWN DOWN NS NS NS 5
GO:0015672 monovalent inorganic cation transportUP UP NS UP NS NS DOWN NS NS NS NS NS NS UP 5
GO:0022900 electron transport chain UP UP NS NS NS NS NS NS DOWN DOWN DOWN NS NS NS 5
GO:0030317 sperm motility UP UP NS NS DOWN DOWN DOWN NS NS NS NS NS NS NS 5
GO:0033539 fatty acid beta-oxidation using acyl-CoA dehydrogenaseNS NS NS NS NS NS NS DOWN DOWN DOWN DOWN DOWN NS NS 5
GO:0035282 segmentation NS DOWN NS NS NS NS NS UP UP UP UP NS NS NS 5
GO:0044710 single-organism metabolic process NS NS NS DOWN NS NS NS NS DOWN DOWN DOWN NS DOWN NS 5
GO:0051186 cofactor metabolic process NS NS NS DOWN NS NS NS DOWN DOWN DOWN DOWN NS NS NS 5
GO:0055085 transmembrane transport UP UP NS UP NS NS DOWN NS NS NS NS UP NS NS 5
Genes Appearing in Multiple Gene Ontology Categories
Pertaining to Lipid Metabolism
Analyzing genes that appear in all of the classes as (i) cellular processes of fatty
acid oxidation, (ii) fatty acid beta oxidation, (iii) lipid oxidation or (iv) fatty acid
beta oxidation using acyl CoA dehydrogenase, in both Li- and HCy-exposed heart
tissue, the genes appearing in all classes included:
acyl CoA dehydrogenase 11 (Acad11)
acyl CoA dehydrogenase long chain (Acadl)
acyl CoA dehydrogenase medium chain (Acadm)
acyl CoA dehydrogenase short chain (Acads)
acyl CoA dehydrogenase very long chain (Acadvl) and
electron transferring flavoprotein dehydrogenase (Etfdh)
Notably all of the protein products of these genes localize to the mitochondria.
Lipids essentially serve three critical cell functions: energy storage, structural
components of membranes, and lipid modifications for activity of cell signaling
factors.
Alcohol alters gene expression similarly to that of elevated HCy
exposure: RT-PCR Analyses of the ED 15.5 mouse heart
Alcohol Exposure Homocysteine
Exposure
Importantly, our RT-PCR data
analyzing the alcohol-exposed
fetal heart indicate the fatty
acid metabolism- and folate-
related genes are upregulated
within the embryonic and fetal
heart tissues, seemingly to
overcome folate metabolism-
related deficiency.
Folate protects gene
expression by maintaining
expression close to control
levels.
RA LA
RVLV
Infer. Vena Cava
Umbilical Artery
Ductus Venosus
PLACENTA
HEARTLIVER
Umbilical Vein
Cross-Talk between Utero-Placental and Fetal Circulation
When genes are knock-out only in the placenta, heart defects can arise. If those
placental genes are re-expressed, the cardiac defects are prevented.(Barak et al., 1999; Adams et al., 2000)
Heart-Placenta Axis: IUGR and Other Effects
Oil Red O staining for neutral lipids (triglycerides, diacylglycerols, and cholesterol esters)
Organization of the labyrinth layer is significantly altered with alcohol exposure
and lipid synthesis is decreased. Folate supplementation returns cell organization and
lipid metabolism nearer to control levels.
Heart-Placenta Axis: IUGR and Ethanol Effects
NMHC-IIA has a unique, but necessary
role in placentation. Function is unknown.
If knocked out, embryonic lethality occurs.
NMHC-IIB appears to be involved in
cell motility. If knocked out, embryos
develop, but die from cardiac defects at
mid-gestation.
Han, et al., 2012. Am J Obstet Gynecol 207, 140 e147-119.
Changes in Fatty Acid Metabolism within the Embryo:
An emerging common theme in...
• Alcohol abuse
• Diabetes
• Obesity
• Lithium exposure
• Elevated homocysteine (folate deficiency)
• ...and they all relate to a higher risk of birth defects
Genes involved in fatty acid metabolism can be epigenetically regulated by specific
methylation patterns within promoter and intragenic regions.
These patterns can be transmitted from one generation to another.
In conclusion…
How much alcohol is safe to drink during
pregnancy?
• No known amount of alcohol is safe for a
growing embryo and fetus.
In the United States 1 in 5 of over half a million women
who report drinking alcohol during pregnancy also admit
to binge drinking. It is known that even low levels of
alcohol (ethanol; EtOH) prenatally can produce human
birth defects.
Severity of defects depends on dose and timing of
alcohol intake and seemingly gender of fetus.
Critical Periods in Human Prenatal Period
2008 2009 2010 2011 2012 2013
EU (28
countries)5469434 5412572 5411129 5266162 5231177
5075691 (e
p)
EU (27
countries)5425681 5367995 5367768 5224965 5189406
5035752 (e
p)
Euro area
(18
countries)
3492593 3435284 3442269 3363927 33195573244394 (e
p)
Euro area
(17
countries)
3468196 3413240 3422488 3345102 32996603223798 (e
p)
Belgium 127205 127198 130100 128705 128051 125606 (p)
Bulgaria 77712 80956 75513 70846 69121 66578
Czech
Republic119570 118348 117153 108673 108576 106751
Denmark 65038 62818 63411 58998 57916 55873
Germany 682514 665126 677947 662685 673544 685000 (e)
Estonia 16028 15763 15825 14679 14056 13531
Ireland 75173 75554 75174 74033 72225 68930 (p)
Greece 118302 117933 114766 106428 100371 94146 (p)
Spain 518503 493717 485252 470553 453348 424494 (p)
France 829311 825564 833654 824263 821844 810820 (e)
Live births are the births of
children that showed any
sign of life (total births minus
stillbirths).Additional information on Eurostat web
page
Live Births In Estonia and Other EU Countries:
What could drinking
during pregnancy mean
for Estonia in relation to
livebirths?
If one considers FASD occurs about one in
every 100 live births (May et al., 2009),
the prevalence of FASD in Estonia
between 2008-2013 (6 years) would be
expected to be ~899 individuals, with
most having varying degrees of
developmental disabilities, with ~54%
(495) possibly having some type of cardiac
defect, or many more being among the
stillbirths.
Mis tähendaks alkahooli joomine raseduse ajal
Eesti ühiskonnale?
Genes Appearing in Multiple Gene Ontology Categories
Pertaining to Lipid Metabolism
Analyzing genes that appear in all of the classes as (i) cellular processes of
fatty acid oxidation, (ii) fatty acid beta oxidation, (iii) lipid oxidation or
(iv) fatty acid beta oxidation using acyl CoA dehydrogenase, in both Li-
and HCy-exposed heart tissue, the genes appearing in all classes included:
acyl CoA dehydrogenase 11 (Acad11)
acyl CoA dehydrogenase long chain (Acadl)
acyl CoA dehydrogenase medium chain (Acadm)
acyl CoA dehydrogenase short chain (Acads)
acyl CoA dehydrogenase very long chain (Acadvl) and
electron transferring flavoprotein dehydrogenase (Etfdh)
Notably all of the protein products of these genes localize to the
mitochondria.
Lipids essentially serve three critical cell functions: energy storage,
structural components of membranes, and lipid modifications for
activity of cell signaling factors.
Alcohol alters gene expression similarly to that of
elevated HCy exposure: RT-PCR Analyses of the ED
15.5 mouse heart
Alcohol Exposure Homocysteine
Exposure
Importantly, our RT-PCR
data analyzing the alcohol-
exposed fetal heart indicate
the fatty acid metabolism-
and folate-related genes are
upregulated within the
embryonic and fetal heart
tissues, seemingly to
overcome folate metabolism-
related deficiency.
Folate protects gene
expression by
maintaining expression
close to control levels.
RA LA
RVLV
Infer. Vena Cava
Umbilical Artery
Ductus Venosus
PLACENTA
HEARTLIVER
Umbilical Vein
Cross-Talk between Utero-Placental and Fetal Circulation
When genes are knock-out only in the placenta, heart defects can arise. If
those placental genes are re-expressed, the cardiac defects are prevented.(Barak et al., 1999; Adams et al., 2000)
Heart-Placenta Axis: IUGR and Other Effects
Oil Red O staining for neutral lipids (triglycerides, diacylglycerols, and cholesterol esters)
Organization of the labyrinth layer is significantly altered with alcohol exposure
and lipid synthesis is decreased. Folate supplementation returns cell
organization and lipid metabolism nearer to control levels.
Heart-Placenta Axis: IUGR and Ethanol Effects
NMHC-IIA has a unique, but necessary
role in placentation. Function is
unknown. If knocked out, embryonic
lethality occurs.
NMHC-IIB appears to be involved in
cell motility. If knocked out, embryos
develop, but die from cardiac defects at
mid-gestation.
Han, et al., 2012. Am J Obstet Gynecol 207, 140 e147-119.
Changes in Fatty Acid Metabolism within the
Embryo: An emerging common theme in...
� Alcohol abuse
� Diabetes
� Obesity
� Lithium exposure
� Elevated homocysteine (folate deficiency)
� ...and they all relate to a higher risk of birth
defectsGenes involved in fatty acid metabolism can be epigenetically regulated
by specific methylation patterns within promoter and intragenic regions.
These patterns can be transmitted from one generation to another.
In conclusion/
How much alcohol is safe to drink during
pregnancy?
� No known amount of alcohol is safe for a
growing embryo and fetus.
In the United States 1 in 5 of over half a million
women who report drinking alcohol during pregnancy
also admit to binge drinking. It is known that even low
levels of alcohol (ethanol; EtOH) prenatally can
produce human birth defects.
Severity of defects depends on dose and timing of
alcohol intake and seemingly gender of fetus.
Critical Periods in Human Prenatal Period
2008 2009 2010 2011 2012 2013
EU (28
countries
)
5469434 5412572 5411129 5266162 52311775075691 (
ep)
EU (27
countries
)
5425681 5367995 5367768 5224965 51894065035752 (
ep)
Euro
area (18
countries
)
3492593 3435284 3442269 3363927 33195573244394 (
ep)
Euro
area (17
countries
)
3468196 3413240 3422488 3345102 32996603223798 (
ep)
Belgium 127205 127198 130100 128705 128051125606 (p
)
Bulgaria 77712 80956 75513 70846 69121 66578
Czech
Republic119570 118348 117153 108673 108576 106751
Denmark 65038 62818 63411 58998 57916 55873
Germany 682514 665126 677947 662685 673544685000 (e
)
Estonia 16028 15763 15825 14679 14056 13531
Ireland 75173 75554 75174 74033 72225 68930 (p)
Greece 118302 117933 114766 106428 100371 94146 (p)
Spain 518503 493717 485252 470553 453348424494 (p
)
France 829311 825564 833654 824263 821844810820 (e
)
Live births are the births of
children that showed any
sign of life (total births
minus stillbirths).Additional information on Eurostat web
page
Live Births In Estonia and Other EU Countries:
What could drinking
during pregnancy mean
for Estonia in relation to
livebirths?
If one considers FASD occurs about
one in every 100 live births (May et al.,
2009), the prevalence of FASD in
Estonia between 2008-2013 (6 years)
would be expected to be ~899
individuals, with most having varying
degrees of developmental disabilities,
with ~54% (495) possibly having
some type of cardiac defect, or many
more being among the stillbirths.
Mis tähendaks alkahooli joomine raseduse
ajal Eesti ühiskonnale?
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