Aboriginal FASD and CNP Workshop Toronto, 6th. March, 2013

Aboriginal FASD and CNP Workshop
Toronto, 6th. March, 2013

Slide – 1
I was very pleased when I was told that you wanted the topic to be epigenetics - it indicated that you
were moving on from knowing about the clinical presentations of FASD. I have urged others here in
Ontario, to address FASD and Epigenetics, without success. As far as I know this is the first conference
that it has been presented. I congratulate you on your insight.
A search of the internet showed that up to now there have been two conferences on epigenetics in
Ontario. One was about epigenetics in general and the other related to epigenetics and cancer. In both
cases there was no indication that the role of alcohol or FASD was considered.
I have made many presentations on FASD. This is my first on epigenetics and it has been the most
difficult to do. I am not a geneticist or a neurologist, and no doubt like you, I find publications on
epigentics difficult to understand. We must not let this prevent us from learning and commenting on
its significance. To do this we have to pick out the information relevant to all of us and which we can
understand, because the implications of epigenetics are very significant.
This presentation will give a brief overview of the history of alcohol, the impact of epigenetics and its
relevance to FASD, a brief review of DNA/RNA, review of some of the areas of the brain and how PAE
effects their function. It will move on to describe the process of epigenetics , methylation and
acetylation. Lastly the brain as a complex system and the unintended consequences of epigenetic
intervention are discussed with final conclusions.

Slide – 2
Prenatal - Traditionally the term “prenatal” covers the moment of fertilization to birth. It is now
apparent that through epigenetics the sperm of the father may contribute to developmental
abnormalities which may show in the child or adult. In addition the epigenetic effects on the male and
female may be passed on to future generations. Consequently we require a word to cover this
generational effect of alcohol. Therefore, in this presentation, prenatal will mean all the generations
preceding any one pregnancy, and prenatal alcohol exposure will mean the immediate alcohol
exposure of the one pregnancy plus all the epigenetic effects of alcohol consumed by both parents in
all the previous generations.

Phenotype – This word is used although the requirements are not met. We see this in the DSM, more
so in the past when the heredity of mental illness was suspected but unproven.
Often the word is used as requiring a set of characteristics unique to one condition although that is not
actually the definition. This requirement of uniqueness is used to argue that FASD and ARND are not
diagnoses, although ARND and FASD fulfill the requirements for a Phenotype.

Epigenetics – Genes have functions, they determine everything, from the color of our eyes to the
intricate and delicate function of brain cells. These functions are called gene expressions.
Until recently it was thought that only gross changes of genes i.e. mutations - changes of DNA
sequence within the gene - could cause changes in gene expression.
We now understand that gene expression can change without changes of DNA sequence. This process
is referred to as epigenetic changes of gene expression.

Slide – 3
Alcohol – There is archeological evidence that alcohol has been available for 12,000 years, in all
ancient communities. References to alcohol have been recorded by all communities and civilizations

over the last 4,000 years. Such records have consistently described its use in religious ceremonies,
and what are seen as its benefits. In addition these records have consistently referred to the disruptive
consequences of alcohol on the individual and society. Nothing has changed over all these centuries.
The descriptions of the disruptive effect of alcohol over recorded history have described the effect of
PAE on the fetus and offspring, but only to a relatively lesser degree. Never the less those references
have been recorded in all societies.
If we take that a new generation in humans occurs every twenty years, we all will have approximately
600 hundred previous generations over the last 12,000 years and 200 over the last 4,000 years.
Each of these generations had the potential to pass on the epigenetic effects of alcohol.
The questions are what kind of humans would we be, what kind of societies would we have, if there
had never been any prenatal alcohol?

Slide – 4
Epigenetics, Alcohol and FASD
Increasingly from the 17th.century references were made to alcohol effecting the offspring. Those
accounts would no doubt not satisfy the rules of research today. Never the less those repeated
accounts pointed the way to our description of FASD today.
In 1899 Dr W. C. Sullivan, a prison doctor, showed that of 6oo children born of 120 alcoholic families
55% died before the age of two. He showed that when the mother drank the child had an increased
chance of being abnormal than when the father drank.
He also showed that when the same mothers were confined in prison for the duration of their
pregnancy, and could not drink alcohol, their offspring were not effected in the way they were when
the same mothers drank out of prison.
In 1912 Dr Charles Stockard showed that chickens, fish and guinea pigs exposure to alcohol prior to
mating sired offspring with defective nervous systems, and that this applied to both parents.- to the
male prior to fertilization and to the female during the pregnancy. In addition he showed that those
defects became less in succeeding generations if there was no further exposure to alcohol. Both of
these observations were subsequently ignored by researchers. Perhaps now with the progress of
epigenetics more attention will be directed to these two significant observations by Stockard.

Slide – 5
Alcohol - 2013
So where are we today. The situation regarding PAE and FASD is the same as it has been over the
centuries. It true that more research is being done but they are still the poor cousins. Any worker or
parent who can compare the time and money given to PAE and FASD to that given to other conditions
will understand what I am saying.
Reiview of pubmed – see slide.
It is difficult to find statistics re. money spent . However, it is apparent that the amount of research and
money spent on alcoholism as it effects the adult has to be in the multiple millions per annum. Far
more than is spent on PAE. This would not be so bad if the drug and alcoholism research community
acknowledged PAE- but they rarely do. Here in Canada, CAMH has no treatment or diagnostic
programs specifically for FASD.
In the U.S.A. for 2012 the National Institutes of Health estimates of funding for Alcoholism was $452
million and $34 million for FAS.
One would hope that now we understand the universal epigenetic effect of alcohol more research
would be applied to PAE. A review of conferences on epigenetics sadly shows the same pattern. PAE is
very much a poor cousin. A list of 23 conferences for 2013 contains no reference to PAE or FASD,
similarly for 2012.

A list of 10 conferences on epigenetics showed no reference to PAE or FASD.
This is a matter of great concern considering that in the real world PAE is possibly the main cause
of dysfunctional changes in gene expression.
Slide – 6
Genetics – DNA. A gene is a group of DNA molecules that have a recognizable hereditary function.
Deoxyribonucleic acid (DNA) contains the characteristics of species and individuals that are passed
on to future generations. This DNA information is passed on at the time of reproduction i.e. when a cell
divides [ replicates ]. The DNA that is passed on remains the same with the same function- unless this
process is damaged [ mutation ] e.g. as by radiation, when the order [ sequence ] of the DNA molecules
is changed.
The DNA also passes on instructions to other cells, telling them what they should do. e.g. produce
more of a certain neurotransmitter.
DNA is located in the nucleus of the cell. A smaller number of DNA molecules is in RNA.
In order to contain the many DNA molecules in the small nucleus they are arranged in tight spiral
formation, rather like a metal spring is reduced in length by have many coils.
Each complete set of DNA molecules, arranged in this tight formation is called a chromosome.
The chromosome unwinds when cells divide and the DNA has to be copied and passed on.
The chromosome also unwinds when the DNA gives its instructions to other cells, throughout the life
of the person. Humans have 22 pairs of chromosomes plus one pair of sex chromosomes that
determine our sex.
The chromosome, with all its DNA molecules, is kept in it spiral form by a special proteins called
Chromatin. Chromatin actually plays a part in activating and directing the specific action of each set of
DNA molecules that make up each gene.
All the DNA of a person is together called the Genome.
The order [ sequence ] in which the DNA collections of Genes are arranged in the chromosome is now
known, but the exact number of genes has not been determined- opinions vary from 20,000 to 75,000.
So sequences of DNA which we call genes provide the instructions that we need in order to reproduce
and develop from a fetus, to a child and adult, including our sex. The DNA instructions also help us to
deal with the environment we live in.
We are built with protein. Protein is formed from the amino acids in our diet. The way the different
proteins are formed to provide all the individual and specialized parts of our body and body function
is determined by the DNA in our genes.
These instructions in the DNA are passed on to their destination [i.e. other cells] through RNA.
Together DNA and RNA are called Nucleic Acids.

[National Human Genome Research Institute ][U.S.]
“DNA is made of chemical building blocks called nucleotides. These building blocks are made of three
parts: a phosphate group, a sugar group and one of four types of nitrogen bases. To form a strand of
DNA, nucleotides are linked into chains, with the phosphate and sugar groups alternating.The four
types of nitrogen bases found in nucleotides are: adenine (A), , thymine (T), guanine (G) and cytosine
(C). The order, or sequence, of these bases determines what biological instructions are contained in a
strand of DNA. For example, the sequence ATCGTT might instruct for blue eyes, while ATCGCT might

instruct for brown. Each DNA sequence that contains instructions to make a protein is known as a
gene. The size of a gene may vary greatly, ranging from about 1,000 bases to 1 million bases in
humans.The complete DNA instruction book, or genome, for a human contains about 3 billion bases
and about 20,000 genes on 23 pairs of chromosomes.”

Slide – 7
Genetics RNA. -RNA is a large molecule that enables any cell to take the DNA instructions and use
them to create proteins [ groups, sequences, of amino acids ] that are special for that living cell. It acts
in the coding, decoding, regulation and expression of the cell and all the cells that replicate from it.
Unlike DNA, which is a double helix [spiral] of nucleotides, RNA is a single spiral of nucleotides.
Messenger RNA [mRNA] carries the instructions from the DNA to the Ribosome where proteins,
specific to that cell, are produced [ protein synthesis, translation]
Transfer RNA [tRNA ] transfers the amino acids to the Ribosome.
RNA in the Ribosome [rRNA ]combines the instructions with the amino acids to form the protein.
There are other forms of RNA with specific roles.
RNA is far less stable than DNA and can be altered with positive and negative consequences.
In viruses RNA can carry genetic instructions and reverse transcription also occurs.
Sets of DNA molecules [ genes] are found in the Mitochondria of our cells. RNA molecules are
transferred to the Mitochondria.
The DNA in the Mitochondria is only inherited from the mother because during the process of
fertilization the male Mitochondria lose their DNA.
Mitochondria provide create and transfer energy for the cells. They are also play important roles in
the cell cycle, differentiation, growth and death. Other functions are storage of Calcium Ions,
regulation of membrane potential, apoptosis, calcium signaling, regulation of cellular metabolism and
steroid synthesis.

Slide 8
Before and Now- Before the explosion of epigenetics –
-PAE meant the drinking of alcohol during the pregnancy, from conception to birth.
- Only the mother was considered to be responsible for the effects of PAE which we term FASD, FAS,
ARND.
- The processes of fertilization, implantation and early gastrulation were considered not to be effected
by alcohol. [ PAE ]
- The father was not thought to contribute to the effects of alcohol [PAE] on the developing fetus and
child.
- Our knowledge and assessments of FASD was based upon the clinical presentation and psychological
assessments, together with gross anatomy and traditional imaging.

While these remain important

Since the explosion of epigenetics and the development of more advanced forms of research –
-It is now known that alcohol [PAE] effects the sperm prior to fertilization, implantation and
throughout gastrulation.
- The consequences of alcohol consumption can no longer be solely “blamed” on the mother.
- Alcohol consumption by the father, prior to conception, also contributes to its deleterious effect on
the fetus and child.

- The definition of PAE needs to include all alcohol consumed by both parents in all preceding
generations.
- The indications are that these developments are not receiving the attention that is so necessary for
the future of the individual and society in general.

Slide –9
Some of the regions of the brain affected by PAE. - The evidence that PAE affects many parts of the
brain and brain function is overwhelming, but largely ignored. There are non so blind as those who
choose not to see. When we review these regions we need to keep in mind the behavior and
difficulties of the children in our care and see how the one explains the other.
Plasticity refers to the ability of the brain to adapt and compensate to injury and environmental
insults, and to positive processes. It can be seen as the positive side of epigenetics.
It is a normal continuous process, to varying degrees, throughout life.
The question we need to ask for each person affected by PAE is – has this person’s brain the capacity
to compensate to the degree that it can be demonstrated both on neuropsychological examination and
in the person’s daily affairs i.e. executive and adaptive functioning?
At this time the answer is mostly no to this question, with the exceptions of intensive sensory motor
integration and exercise for the infant. Certain vitamins have a similar affect at this age.
As always with FASD and PAE there are unintended consequences.
Changing the affects of PAE from “more affected to less affected” brings different problems.
The more intelligent person with FASD suffers in different ways. Negative accusations are made
against them when they are not able to reach or maintain the standards expected of them. Also,
because of their greater abilities they get into more complicated negative situations that bring them
into conflict with the law etc.
If a region of an animal brain is exposed to alcohol and as a result that animal acts violently towards
others would we say it chose to behave that way- no we would not. So why do we hold humans
responsible for similar behavior when they have the brain damage of PAE?

Slide – 10
Brain size and cortical thickness – these are two simple but significant observations.
Brain [head] size is used in the screening and diagnosis of FASD.
They both represent the gross impact of PAE on the developing brain. It is important to note however
that normal brain size does not exclude serious neurological dysfunction and the diagnosis of FASD
[ARND]
Small brain size is associated with decreased white and gray matter, white more than gray, and
abnormalities of the cerebellum, basal ganglia and corpus callosum associated with deficits of the
overall I.Q., executive and motor functioning. Not only is it significant that both are reduced but the
fact that the ratio of one to another is different to normal may be of significance.
White matter relays and coordinates communication between gray, nerve cell bodies, in different
parts of the brain. It consists of bundles of axons which are covered in myelin. The myelin insulates
the axon and increases the speed of transmission along the axon. These myelinated axons [ seen
grossly as white matter] connect gray matter [ cell bodies ] from cortex to cortex, hemisphere to
hemisphere as well as other structures one to another.
Besides the cell bodies gray matter includes the cell dendrites and Glial cells. Glial cells are not nerve
cells. They give physical support and regulate the fluid surrounding their adjacent nerve cells. In this
way they regulate the synapses between cells and cell nutrition. They are responsible for the
migration of neurons, modify the growth of axons and dendrites, and contribute to the regulation of
synaptic function. They also have an important role in the plasticity and repair.

The small size is not the cause of these findings but rather the gross sign of their presence as a
consequence of PAE – just as chemical imbalance is not the cause of mood disorders of the DSM. For
years, and is still the case for some, we have made a “small head” and “chemical imbalance” as the
cause and therefore no need to look any further.

Slide – 11
CORPUS CALLOSUM– From Wikipedia, The posterior portion of the corpus callosum is called the splenium; the anterior is
called the genu (or "knee"); between the two is the truncus, or "body", of the corpus callosum. The part between the body and the
splenium is often markedly thinned and thus referred to as the "isthmus". The rostrum is the part of the corpus callosum that
projects posteriorly and inferiorly from the anteriormost genu, as can be seen on the sagittal image of the brain displayed on the
right. The rostrum is so named for its resemblance to a bird's beak. Thinner axons in the genu connect the prefrontal cortex
between the two halves of the brain. Thicker axons in the midbody of the corpus callosum and in the splenium interconnect areas
of the premotor and supplementary motor regions and motor cortex, with proportionally more corpus dedicated to supplementary
motor regions. The posterior body of the corpus communicates somatosensory information between the two halves of the parietal
lobe and the visual center at the occipital lobe.

The Corpus Callosum lies between the two hemispheres of the brain. It consists of tracts or bundles of
white matter, i.e. myelinated axons, that arise from the cell bodies of one hemisphere and pass to the
other hemisphere where they communicate with cell bodies of the second hemisphere.
The Corpus Callosum was one of first major structures of the brain to be identified as damaged by
PAE. Because it is related to all parts of each hemisphere it is expected that the effects will be varied.
Agenesis occurs in extreme cases- 6.8% in children exposed to prenatal alcohol compared to 0.3% in
the general population.
The posterior region is most affected on imaging. Variations of shape occur in the absence of changes
in size, in association with impairments of executive functioning and motor deficits. Other functions of
the brain that are affected may include verbal learning, bimanual coordination, accuracy of
interpretation, visual and auditory memory.
With the more advanced diffusion tensor imaging it has been shown that these changes, with
impairments of function, can occur in the absence of the facial features of FAS.

Slide – 12
Amygdalae – They are part of the limbic system and include a number of nuclei with different
functions. The Amygdala and Pyriform Cortex are significant for processing memory related to
emotional events and control and learning of emotional and social behaviors [ adaptive functioning ].
Abnormal, excessive apoptosis occurs with PAE in these regions.
Other functions involve the olfactory bulb and cortex [smell], the activation of the sympathetic system
[ hypothalamus ] and fear response. Pavlovian fear conditioning is impaired. Similarly positive
conditioning can be impaired.
The Amygdala is not a long term memory site but does modulate and regulate memory consolidation
in the other memory sites.
Clinical behavior associated with Amygdalae dysfunction include overreaction, hypoemotionality,
[Alexithymia], loss of fear, difficulty with facial recognition, impaired inhibitory avoidance,
hypersexuality, hyperorality, reduction in maternal instinctive behavior, anxiety disorders, obsessive
compulsive disorders, post traumatic stress disorder, borderline personality disorder.
Variations in size of the right and left Amygdalae are associated with some of these dysfunctions.
Alternative sexual orientation is related to differences in the Amygdalae.

The Amygdalae are also involved with aggression. Stimulation of the Amygdalae decreases aggression
while brain lesions involving the Amygdalae increases aggression. Alcoholism is associated with
reduced activity of the Amygdala.

Publications
- Epigenetic alterations are critical for fear memory consolidations and synaptic plasticity in the
lateral Amygdala.- Melissa S. Monsey,1 Kristie T. Ota,1 Irene F. Akingbade,1 Ellie S. Hong,1 and Glenn E. Schafe1,2,*
The role of alcohol was not mentioned.

- Low Dose Prenatal Ethanol Exposure Induces Anxiety- Like Behaviour and Alters Dendritic
Morphology in the Basolateral Amygdala of Rat Offspring
Carlie L. Cullen1, Thomas H. J. Burne2, Nickolas A. Lavidis1, Karen M. Moritz1*
This study showed that low doses of PAE caused detectible changes in the Amygdala of rats associated
with increased anxiety like behavior.
-
Sexually Dimorphic Effects of Alcohol Exposure during Development on the Processing of Social Cues
Sandra J. Kelly,* Darnica C. Leggett, and Kim Cronise
This study shows that PAE interferes with recognition memory in both sexes although there are differences
which are thought to be due to the influence of oxytocin in the female. In females the oxytocin receptor
binding in the amygdala was reduced by PAE.
Slide – 13
Thalamus – The Thalamus is a bi-lobed midline structure on top of the brain stem. It receives sensory
information from the spinal cord about pain, temperature and touch. It has connections to the
hippocampus.
All sensory systems, except olfactory, are relayed to their appropriate cortical areas.
The Thalamus also helps regulate sleep, wakefulness, arousal and level of awareness. The Thalamus
also contributes to spatial, recollective and familiarity memory.

Publications
Brain diffusion abnormalities in children with FASD Lebel C, Rasmussen C, Wyper K, Walker L, Andrew
G, Yager J, Beaulieu C.
-“Diffusion tensor imaging revealed significant differences of diffusion parameters in several areas
of the brain, including the genu and splenium of the corpus callosum, cingulum, corticospinal
tracts, inferior fronto-occipital fasciculus, inferior and superior longitudinal fasciculi, globus pallidus,
putamen, and thalamus. Reduced white and gray matter volumes, as well as total brain volume,
were observed in the FASD group.”
Brain metabolic alterations in adolescents and young adults with fetal alcohol spectrum
disorders.
Fagerlund A, Heikkinen S, Autti-Rämö I, Korkman M, Timonen M, Kuusi T, Riley EP, Lundbom N.
In patients with FASD, lower NAA/Cho and/or NAA/Cr compared with controls were found in
parietal and frontal cortices, frontal white matter, corpus callosum, thalamus, and cerebellar
dentate nucleus. There was an increase in the absolute intensity of the glial markers Cho and Cr
but no change in the neuronal marker NAA.

Extensive deep gray matter volume reductions in children and adolescents with fetal
alcohol spectrum disorders.
Nardelli A, Lebel C, Rasmussen C, Andrew G, Beaulieu C.

Significant reductions of volume in FASD were observed for the intracranial vault (7.6%), total white matter
(8.6%), total cortical gray matter (7.8%), and total deep gray matter (13.1%). All 6 deep gray matter
structures showed significant volume reductions bilaterally with the caudate (approximately 16%) and
globus pallidus (approximately 18%) being most affected. The hippocampus, thalamus, and globus pallidus
showed reductions in all 3 age subgroups (6 to 9, 10 to 13, and 14 to 17 years) but the caudate and putamen
had smaller volumes for FASD only within the 2 youngest subgroups; the amygdala was only smaller for
FASD in the 2 oldest subgroups.
Slides – 14 -15
Hypothalamus – connects the nervous system to the endocrine system through the pituitary gland.
It controls hunger, thirst, body temperature, sleep, fatigue and the circadian cycle. It also has a role
in parenting and attachment. It produces and secretes neurohormones that can stimulate or inhibit
pituitary hormones. Unlike the rest of the brain the hypothalamus is directly sensitive to hormones
in the blood, such as thyroid hormones.
The Hypothalamus is grouped with the pituitary and adrenal glands to form the HPA Axis.
Most studies relate to this Axis. Dysfunction of this Axis affects the immune system.
Publications
Prenatal alcohol exposure: fetal programming and later life vulnerability to stress,
depression and anxiety disorders. Hellemans KG, Sliwowska JH, Verma P, Weinberg J.
the research suggests that the stress-diathesis model provides a powerful approach for elucidating
mechanisms underlying the increased vulnerability to mental illness [ depression/anxiety ] among
individuals with FASD, and developing appropriate treatments for these individuals.
Prenatal alcohol exposure: foetal programming, the hypothalamic-pituitary-adrenal axis
and sex differences in outcome. Weinberg J, Sliwowska JH, Lan N, Hellemans KG.
Here, we review data demonstrating that alcohol exposure in utero programmes the foetal
HPA axis such that HPA tone is increased throughout life. Importantly, we show that,
although alterations in HPA responsiveness and regulation are robust phenomena,
occurring in both male and female offspring, sexually dimorphic effects of alcohol are
frequently observed.
Prenatal alcohol exposure and fetal programming: effects on neuroendocrine and immune
function. Zhang X, Sliwowska JH, Weinberg J.
We present data that demonstrate that maternal alcohol consumption increases HPA activity in
both the maternal female and the offspring. Increased exposure to endogenous glucocorticoids
throughout the lifespan can alter behavioral and physiologic responsiveness and increase
vulnerability to illnesses or disorders later in life. Alterations in immune function may be one of the
long-term consequences of fetal HPA programming. We discuss studies that demonstrate the
adverse effects of alcohol on immune competence and the increased vulnerability of ethanol-
exposed offspring to the immunosuppressive effects of stress. Fetal programming of HPA activity
may underlie some of the long-term behavioral, cognitive, and immune deficits that are observed

following prenatal alcohol exposure.
Neuroendocrine effects of fetal alcohol exposure.
Taylor AN, Branch BJ, Kokka N. [1981]
Our studies of the long-term effects of fetal and early postnatal exposure to alcohol on pituitary-
adrenal and body temperature responses to a challenge dose of ethanol in adult rats are
described. Both responses are enhanced in prenatally, but not in postnatally exposed rats,
indicating that the effects of fetal alcohol exposure on physiological systems, such as the
endocrine and thermoregulatory systems, persist to adulthood.
Slide – 16
Hippocampus – The Hippocampus is part of the cerebral cortex. It is present in both hemispheres.
It plays its role in short term and long term memory, known as episodic memory. Extensive loss of
the Hippocampi results in an inability to form new memories or recall memories of events after the
damage, but memories from before the damaged are recalled. Other functions are spatial
orientation, possibly inhibition and learning- all related to cell loss and impaired neurogenesis.
Imaging studies have found varying degrees of alterations of morphology of the Hippocampus with
PAE.
Publications
Persistent impairment of hippocampal neurogenesis in young adult rats
following early postnatal alcohol exposure.
Klintsova AY, Helfer JL, Calizo LH, Dong WK, Goodlett CR, Greenough WT.
CONCLUSIONS: These observations suggest that early postnatal binge alcohol exposure results
in long-term deficits of adult hippocampal neurogenesis, providing a potential basis for the deficits
of hippocampus-dependent behaviors reported for this model.
GABAergic Gene Expression in Postmortem Hippocampus from Alcoholics and Cocaine Addicts;
Corresponding Findings in Alcohol- Naïve P and NP Rats
Mary-Anne Enoch , Zhifeng Zhou, Mitsuru Kimura, Deborah C. Mash, Qiaoping Yuan, David
Goldman
Our study confirms the involvement of the GABAergic system in alcoholism but also reveals a
hippocampal GABA input in cocaine addiction. Congruent findings in human addicts and P rats
provide clues to predisposing factors for alcohol and drug addiction. Finally, the results of this
study have therapeutic implications.
Slide – 17
Prefrontal Cortex – The Prefrontal Cortex acts in modulation of social behavior, expression of
personality, planning and decision making i.e. it has an important contribution to executive
functioning.
Impaired executive functioning is universal, to varying degrees, in FASD. Impairment is
independent of overall intellectual functioning [ I.Q. ]
We will all recognize, by its absence, the following description of EF.
From Wikipedia- “The most typical psychological term for functions carried out by the prefrontal

cortex area is executive function. Executive function relates to abilities to differentiate among
conflicting thoughts, determine good and bad, better and best, same and different, future
consequences of current activities, working toward a defined goal, prediction of outcomes,
expectation based on actions, and social "control" (the ability to suppress urges that, if not
suppressed, could lead to socially unacceptable outcomes).”
PAE is associated with morphological changes to the cell dendrites of the Prefrontal Cortex and
cell loss in rodents.
Publications
Neuronal reduction in frontal cortex of primates after prenatal alcohol exposure.Burke MW,
Palmour RM, Ervin FR, Ptito M.
Children with fetal alcohol spectrum disorders (FASD) show behavioral and intellectual
impairments that indicate frontal lobe dysfunction, but the extent of damage to this region has not
been clarified by brain imaging studies. This study uses the St Kitts vervet monkey, a species that
voluntarily consumes beverage alcohol, to examine the effects of prenatal ethanol exposure.
Pregnant vervets were allowed to drink the equivalent of 3-5 standard drinks four times a week
during the third trimester. Using unbiased stereology, we estimated neuronal reduction and found
significantly fewer cells in the frontal lobes of FASD offspring as well as an increased density of
interstitial white matter neurons. These cytoarchitectonic effects are consistent with the behavioral
and cognitive changes observed in FASD.
Prenatal ethanol exposure in rats decreases levels of complexin proteins in the frontal
cortex. Barr AM, Hofmann CE, Phillips AG, Weinberg J, Honer WG.
These data indicate that prenatal exposure to ethanol is associated with a selective loss of
complexin proteins in the frontal cortex. These proteins are known to be important for activity-
dependent neurotransmission, and have previously been shown to mediate synaptic plasticity and
cognition. These combined findings suggest that further study of complexin proteins as a substrate
for cognitive impairment related to prenatal exposure to ethanol is warranted.
Prenatal alcohol exposure affects frontal-striatal BOLD response during inhibitory
control.Fryer SL, Tapert SF, Mattson SN, Paulus MP, Spadoni AD, Riley EP.
These data provide an account of response inhibition-related brain functioning in youth with FASD.
Furthermore, results suggest that the frontal-striatal circuitry thought to mediate inhibitory control is
sensitive to alcohol teratogenesis.
The neural basis of belief encoding and integration in moral judgment.
Young L, Saxe R.
Moral judgment in the mature state depends on "theory of mind", or the capacity to attribute mental
states (e.g., beliefs, desires, and intentions) to moral agents. The current study uses functional
magnetic resonance imaging (fMRI) to investigate the cognitive processes for belief attribution in
moral judgment. Participants read vignettes in a 2x2x2 design: protagonists produced either a
negative or neutral outcome, based on the belief that they were causing the negative outcome or
the neutral outcome; presentation of belief information either preceded or followed outcome
information. In each case, participants judged the moral permissibility of the action. The results
indicate that while the medial prefrontal cortex is recruited for processing belief valence, the

temporo-parietal junction and precuneus are recruited for processing beliefs in moral judgment via
two distinct component processes: (1) encoding beliefs and (2) integrating beliefs with other
relevant features of the action (e.g., the outcome) for moral judgment.
Investigating emotion in moral cognition: a review of evidence from functional
neuroimaging and neuropsychology.
Young L, Koenigs M.
INTRODUCTION:
Human moral decision-making has long been a topic of philosophical debate, and, more recently,
a topic for empirical investigation. Central to this investigation is the extent to which emotional
processes underlie our decisions about moral right and wrong. Neuroscience offers a unique
perspective on this question by addressing whether brain regions associated with emotional
processing are involved in moral cognition.
METHOD:
We conduct a narrative review of neuroscientific studies focused on the role of emotion in morality.
Specifically, we describe evidence implicating the ventromedial prefrontal cortex (VMPC), a brain
region known to be important for emotional processing.
RESULTS:
Functional imaging studies demonstrate VMPC activation during tasks probing moral cognition.
Studies of clinical populations, including patients with VMPC damage, reveal an association
between impairments in emotional processing and impairments in moral judgement and behaviour.
CONCLUSIONS:
Considered together, these studies indicate that not only are emotions engaged during moral
cognition, but that emotions, particularly those mediated by VMPC, are in fact critical for human
morality.
Not all false memories are created equal: the neural basis of false recognition. Garoff-Eaton
RJ, Slotnick SD, Schacter DL.
False recognition, a type of memory distortion where one claims to remember something that
never happened, can occur in response to items that are similar but not identical to previously
seen items (i.e., related false recognition) or in response to novel items (i.e., unrelated false
recognition). It is unknown whether these 2 types of memory errors arise from the same or distinct
neural substrates. Using functional magnetic resonance imaging, we compared the neural activity
associated with true recognition, related false recognition, and unrelated false recognition for
abstract shapes. True recognition and related false recognition were associated with similar
patterns of neural activity, including activity in the prefrontal cortex, the parietal cortex, and the
medial temporal lobe. By contrast, unrelated false recognition was associated with activity in
language-processing regions. These results indicate that false recognition is not a unitary
phenomenon, but rather can reflect the operation of 2 distinct cognitive and neural processes.
Choline supplementation and DNA methylation in the hippocampus and prefrontal cortex of
rats exposed to alcohol during development. Otero NK, Thomas JD, Saski CA, Xia X, Kelly SJ.
This study is the first to show changes in global DNA methylation of the hippocampal region and
PFC after neonatal alcohol exposure. Choline supplementation impacts global DNA methylation in

these 2 brain regions in alcohol-exposed and control animals in a differential manner. The current
findings suggest that both alcohol and choline have substantial impact on the epigenome in the
PFC and hippocampus.
Slide – 18
Cerebellum- The Cerebellum helps modulates motor activity and learning- precision and timing. It
also contributes to attention, language and regulation of emotions.
From Wikipedia - The cerebellum (Latin for little brain) is a region of the brain that plays an
important role in motor control. It may also be involved in some cognitive functions such as
attention and language, and in regulating fear and pleasure responses, but its movement-related
functions are the most solidly established. The cerebellum does not initiate movement, but it
contributes to coordination, precision, and accurate timing. It receives input from sensory systems of
the spinal cord and from other parts of the brain, and integrates these inputs to fine tune motor
activity. Because of this fine-tuning function, damage to the cerebellum does not cause paralysis,
but instead produces disorders in fine movement, equilibrium, posture, and motor learning.
The cerebellum also is necessary for several types of motor learning, the most notable one being
learning to adjust to changes in sensorimotor relationships. Thus, the general conclusion reached
decades ago is that the basic function of the cerebellum is not to initiate movements, or to decide
which movements to execute, but rather to calibrate the detailed form of a movement [modulation ]
Functional imaging studies have shown cerebellar activation in relation to language, attention, and
mental imagery; correlation studies have shown interactions between the cerebellum and non-
motoric areas of the cerebral cortex
There is considerable evidence that the cerebellum plays an essential role in some types of motor
learning. The tasks where the cerebellum most clearly comes into play are those in which it is
necessary to make fine adjustments to the way an action is performed.
Publications
Diffusion Tensor Imaging of the Cerebellum and Eyeblink Conditioning in Fetal Alcohol
Spectrum Disorder
B.S. Spottiswoode, Ph.D.1,2, E.M. Meintjes, Ph.D1, A.W. Anderson, Ph.D.3,4, C.D. Molteno,
M.D.5, M.E. Stanton, Ph.D.6, N.C. Dodge7, J.C. Gore, Ph.D.4,10, B.S. Peterson, M.D.8, J.L.
Jacobson, Ph.D.5,7,9, and S.W. Jacobson, Ph.D.5,7,9
This study extends recent findings that have used DTI to reveal microstructural deficits in white matter in
corpus callosum and throughout the brain in children with FASD to show reduced FA and increased
diffusivity in the cerebellar peduncles, a white matter structure that has been shown in animal models to be
critically important in EBC. The strongest group differences were seen on the perpendicular diffusivity
measure, which suggests poorer axon packing density and/or myelination. The inference of a myelination
deficit is supported by the animal model studies linking prenatal alcohol exposure to impairment in the
oligodendrocytes and expression of myelin basic protein involved in producing the myelin sheath.
Effects of Prenatal Alcohol Exposure on Cerebellum Development The Embryo Project, Arizona
State University
renatal exposure to alcohol (ethanol) results in a continuum of physical, neurological, behavioral,
and learning defects collectively grouped under the heading fetal alcohol spectrum disorders

(FASD). Fetal alcohol syndrome (FAS) is the most severe combination of these defects under this
heading, and is characterized by pre- and postnatal growth deficiencies, facial abnormalities, and
defects of the central nervous system (CNS). The developing brain is particularly vulnerable to the
toxicity of ethanol, given the broad time frame of susceptibility from neurulation, when the neural
tube is formed, all the way through to birth. The cerebellum is an area of the brain particularly
vulnerable to prenatal ethanol exposure. Mechanisms proposed for this drastic reduction in brain
cells include apoptosis, oxidative stress, and damage to the radial glia stem cell progenitor pool.
Physical dexterity, coordination, and visuospatial processing are all affected by these stressors, and
eyeblink classical conditioning tests have proven that ethanol-induced damage goes beyond motor
coordination by permanently impacting learning and memory.
Mechanisms of Ethanol-induced Death of Cerebellar Granule Cells
Jia Luo
The cerebellum is one of the brain areas that is most susceptible to ethanol during development. Ethanol
exposure causes a loss of both cerebellar Purkinje cells and granule cells
The mechanisms of ethanol-induced CGC death are complex and likely reflect the combined outcomes of
promoting intrinsic apoptotic pathways and inhibiting anti-apoptotic signaling. Multiple mechanisms may
interplay; these include inhibition of NMDA receptors, interference with signaling by neurotrophic factors,
induction of oxidative stress, modulation of retinoid acid signaling, disturbance of potassium channel
currents, thiamine deficiency, and disruption of translational regulation.
Resveratrol Restores Nrf2 Level and Prevents Ethanol-Induced Toxic Effects in the
Cerebellum of a Rodent Model of Fetal Alcohol Spectrum Disorders
Ambrish Kumar, Chandra K. Singh, Holly A. LaVoie, Donald J. DiPette and Ugra S. Singh
n humans, ethanol exposure during pregnancy produces a wide range of abnormalities in
infants collectively known as fetal alcohol spectrum disorders (FASD). Neuronal
malformations in FASD manifest as postnatal behavioral and functional disturbances. The
cerebellum is particularly sensitive to ethanol during development. In a rodent model of
FASD, high doses of ethanol (blood ethanol concentration 80 mM) induces neuronal cell
death in the cerebellum. However, information on potential agent(s) that may protect the
cerebellum against the toxic effects of ethanol is lacking. Growing evidence suggests that
a polyphenolic compound, resveratrol, has antioxidant and neuroprotective properties.
Here we studied whether resveratrol (3,5,4′-trihydroxy-trans-stilbene), a phytoalexin
found in red grapes and blueberries, protects the cerebellar granule neurons against
ethanol-induced cell death. In the present study, we showed that administration of
resveratrol (100 mg/kg) to postnatal day 7 rat pups prevents ethanol-induced apoptosis
by scavenging reactive oxygen species in the external granule layer of the cerebellum
and increases the survival of cerebellar granule cells studies indicate that resveratrol
exhibits neuroprotective effects in cerebellum by acting at redox regulating proteins in a
rodent model of FASD.

Teratogenic Effects of Alcohol on Brain and Behavior
Sarah N. Mattson Ph. D. ; Amy M. Schoenfeld; and Edward P. Riley, Ph. D.
erebellum. Another area of the brain that is affected by prenatal alcohol exposure is the
cerebellum, which is involved in both motor and cognitive skill sand is located at the base of the
brain. For example, damage to the cerebellum has been implicated in learning deficits as well as in
balance and coordination, all of which are impaired by prenatal alcohol exposure. A recent study
found that the overall volume of the cerebellum was disproportionately reduced relative to overall
brain size in people with FAS compared with control subjects ( Archibald et al. 2001) . These
findings partially replicate previous reports of reduced cerebellar size in FAS and PEA children (
Sowell et al. 1996) . In addition to the overall reductions in the size of the cerebellum, studies
conducted in both humans and animals suggest that a specific region of the cerebellum the
anterior portion of the cerebellar vermis is particularly affected by
alcohol exposure before or shortly after birth1 (Goodlett et al. 1990; Sowell et al. 1996) (1The
studies in animals, primarily rodents, were conducted shortly after birth, a period that corresponds
to the third trimester of gestation in humans with respect to brain development).
Slide – 19
From Wikipedia - The basal ganglia (or basal nuclei) are a group of nuclei of varied origin in the
brains of vertebrates that act as a cohesive functional unit. They are situated at the base of the
forebrain and are strongly connected with the cerebral cortex, thalamus and other brain areas. The
basal ganglia are associated with a variety of functions, including voluntary motor control,
procedural learning relating to routine behaviors or "habits" such as bruxism, eye movements, and
cognitive,[1] emotional functions. Currently popular theories implicate the basal ganglia primarily
in action selection, that is, the decision of which of several possible behaviors to execute at a given
time. Experimental studies show that the basal ganglia exert an inhibitory influence on a number of
motor systems, and that a release of this inhibition permits a motor system to become active. The
"behavior switching" that takes place within the basal ganglia is influenced by signals from many
parts of the brain, including the prefrontal cortex, which plays a key role in executive functions.
The neurotransmitter dopamine, plays an important role in basal ganglia function.
There are also many indications that it is involved in the control of behavior in a more fundamental
way, at the level of motivation.
Brain dysmorphology in individuals with severe prenatal alcohol exposure.
Archibald SL, Fennema-Notestine C, Gamst A, Riley EP, Mattson SN, Jernigan TL.
The present study used sMRI to examine in detail the regional pattern of brain hypoplasia resulting
from prenatal exposure to alcohol using a higher resolution imaging protocol and larger sample
sizes than reported previously. Fourteen participants (mean 11.4 years; eight females, six males)
with fetal alcohol syndrome (FAS) and 12 participants (mean 14.8 years; four females, eight
males) with prenatal exposure to alcohol (PEA) but without the facial features of FAS were
compared to a group of 41 control participants (mean 12.8 years, 20 females, 21 males). Findings
of significant microcephaly and disproportionately reduced basal ganglia volumes in the FAS
group were confirmed.
Fetal alcohol syndrome and the developing socio-emotional brain.
Niccols A.
Researchers investigating neuropsychological functioning have identified deficits in learning,
memory, executive functioning, hyperactivity, impulsivity, and poor communication and social skills

in individuals with FAS and fetal alcohol effects (FAE). Investigators using autopsy and brain
imaging methods have identified microcephaly and structural abnormalities in various regions of
the brain (including the basal ganglia, corpus callosum, cerebellum, and hippocampus) that may
account for the neuropsychological deficits.
Neuroimaging and Fetal Alcohol Spectrum Disorders
Andrea D. Spadoni,1,* Christie L. McGee,1 Susanna L. Fryer,1 and Edward P. Riley2
additionally, this pattern of abnormalities is thought to be closely related to many of the functional
deficits seen in FASD individuals, for instance, deficits in spatial memory, perseveration, and
difficulty set shifting.
Slide – 20
From Wikipedia - The thymus is a specialized organ of the immune system. The thymus "educates"
T-lymphocytes (T cells), which are critical cells of the adaptive immune system.
Each T cell attacks a foreign substance which it identifies with its receptor. T cells have receptors
which are generated by randomly shuffling gene segments. Each T cell attacks a different antigen. T
cells that attack the body's own proteins are eliminated in the thymus. Thymic epithelial cells
express major proteins from elsewhere in the body, and T cells that respond to those proteins are
eliminated through programmed cell death (apoptosis).
The thymus is largest and most active during the neonatal and pre-adolescent periods. By the early
teens, the thymus begins to atrophy and thymic stroma is replaced by adipose (fat) tissue.
Nevertheless, residual T lymphopoiesis continues throughout adult life.
In the two thymic lobes, hematopoietic precursors from the bone-marrow, referred to as thymocytes,
mature into T-cells. Once mature, T-cells emigrate from the thymus and constitute the peripheral T-
cell repertoire responsible for directing many facets of the adaptive immune system. Loss of the
thymus at an early age through genetic mutation (as in DiGeorge Syndrome) results in severe
immunodeficiency and a high susceptibility to infection.
As the thymus is the organ of T-cell development, any congenital defect in thymic genesis or a
defect in thymocyte development can lead to a profound T cell primary immunodeficiency
Publications
Prenatal Alcohol Exposure and Fetal Programming: Effects on Neuroendocrine and Immune Function
INGQI ZHANG, JOANNA H. SLIWOWSKA, AND JOANNE WEINBERG1
Impaired Immunity in Children with FAS. Children prenatally exposed to alcohol have an increased
incidence of bacterial infections, such as meningitis, pneumonia, otitis media, gastroenteritis, and
sepsis, as well as urinary tract and upper respiratory tract infections (60, 61). Early studies reported
that these children also had lower cell counts of eosinophils and neutrophils, decreased circulating E-
rosette–forming lymphocytes, reduced mitogen-stimulated proliferative responses by peripheral
blood leukocytes, and hypogammaglobulinemia .
Deficits in Cell-Mediated Immunity. Prenatal ethanol exposure alters thymus development in both rats
and mice. Delayed ontogeny of the thymus , decreased total numbers of thymocytes, and diminished
mitogen-induced cell proliferation have been reported in 18- to 19-day-old fetuses . Decreased thymus
weight, size, and cell counts have also been observed at birth . One study in mice found that total
thymocyte numbers returned to control levels as early as postnatal Day 6 (69). However, other studies
reported that changes in thymus measures persist throughout the preweaning period and even into
adolescence . Similarly, mitogen-induced proliferative responses of thymic cells appears to be
suppressed in ethanol-exposed males at weaning but may be increased during the adolescent period.

The adverse effects of ethanol on development of the thymus are confirmed by data from in vitro
studies that used organ culture to assess the direct effects of ethanol.

Slide – 21
Cell Structure, Function and Neurotransmitters
Publications
Effects of prenatal ethanol exposure on voltage-dependent calcium entry into neonatal
whole brain-dissociated neurons.
Lee YH, Spuhler-Phillips K, Randall PK, Leslle SW.
Prenatal ethanol exposure resulted in a significant reduction of calcium entry into K(+)-depolarized
cells, compared with AL and PF control treatments.
Thus, these results suggest that prenatal ethanol exposure decreases voltage-dependent calcium
entry into neonatal- dissociated neurons in a manner that does not seem to involve the selective
inhibition of any individual N-, P-, or L-type calcium channel.

Neural activity and survival in the developing nervous system.
Mennerick S, Zorumski CF.
Recent evidence suggests that blockade of normal excitation in the immature nervous system may
have profound effects on neuronal survival during the period of natural cell death. Cell loss
following depression of electrical activity in the central nervous system (CNS) may explain the
neuropsychiatric deficits in humans exposed to alcohol or other CNS depressants during
development. Thus, understanding the role of electrical activity in the survival of young neurons is
an important goal of modern basic and clinical neuroscience. Here we review the evidence from in
vivo and in vitro model systems that electrical activity participates in promoting neuronal survival.
We discuss the potential role of moderate elevations of intracellular calcium in promoting survival,
and we address the possible ways in which activity and conventional trophic factors may interact.
Synaptic membrane alterations in rats exposed to alcohol.
Guerri C.
These findings indicate that prenatal alcohol exposure probably delays synaptic development,
whereas continued alcohol exposure during lactation may, in addition, alter the physicochemical
structure of synaptic membranes.
Transmembrane Regulation of Intracellular Calcium by a Plasma Membrane
Sodium/Calcium Exchanger in Mouse Ova1
J.R. Pepperell,2,3 K. Kommineni,3 S. Buradagunta,3 P.J.S. Smith,4 and D.L. Keefe3
Intracellular calcium is a ubiquitous element that is implicated in many cell processes, including secretion,
mitosis, and meiosis . Furthermore, sustained elevated intracellular calcium concentrations induce cell death
Hence, all mammalian cells precisely control intracellular levels of calcium during normal cell function.
Slide – 22
Methylation and Acetylation
From Wikipedia- DNA methylation is a biochemical process involving the addition of a methyl
group to the cytosine or adenine DNA nucleotides. DNA methylation stably alters the expression of
genes in cells as cells divide and differentiate from embryonic stem cells into specific tissues. The
resulting change is normally permanent and unidirectional, preventing one organism from reverting
back to a stem cell or converting into another type of tissue. DNA methylation is typically removed

during zygote formation and re- established through successive cell divisions during development.
However, the latest research shows that hydroxylation of methyl groups occurs rather than complete
removal of methyl groups in zygote.
In addition, DNA methylation suppresses the expression of
endogenous retroviral genes and other harmful stretches of
DNA that have been incorporated into the genome of the host over time. DNA methylation also
forms the basis of chromatin structure, which enables a single cell to grow into multiple organs or
perform multiple functions.
methylation can be involved in modification of heavy metals, regulation of gene expression,
regulation of protein function, and RNA metabolism
DNA methylation also plays a crucial role in the development of nearly all types of cancer
DNA methylation is essential for normal development and is associated with a number of key
processes including genomic imprinting, X-chromosome inactivation, suppression of repetitive
elements, and carcinogenesis
Research has suggested that long-term memory storage in humans may be regulated by DNA
methylation.
Methylation contributing to epigenetic inheritance can occur through either DNA
methylation or protein methylation.
Acetylation (or in IUPAC nomenclature ethanoylation) describes a reaction that introduces an
acetyl functional group into a chemical compound. (Deacetylation is the removal of the acetyl
group.)
Acetylation is important in cell (biology) because acetyl groups can turn proteins and genes on and
of
Publications
. Alteration of gene expression by alcohol exposure at early neurulation
Feng C Zhou1*, Qianqian Zhao2, Yunlong Liu2, Charles R Goodlett4, Tiebing Liang2, Jeanette N
McClintick3, Howard J Edenberg3, Lang Li2
In summary, alcohol exposure during the period of early neurulation at ~E8-E10, is predominantly
inhibitory to gene expression, particularly the neural developmental genes. We found major
reductions in gene sets involved in neurospecification, neural growth factors, cell growth and
hematopoiesis. These effects on gene expression parallel the growth delay and developmental
abnormalities including brain, neural tube, eye, heart, blood cells, and embryonic vascularization
which are major targets in FASD. Our study, in conjunction with others that use different
developmental periods of alcohol exposure, provides an important portfolio of alcohol-induced
changes in gene expression associated with altered development. Together, these gene profiles should
contribute to the generation of testable new hypotheses concerning the mechanistic path from gene
expression changes to embryonic structural deficits, and for causal mechanisms of alcohol-induced
teratogenesis (e.g., brain growth retardation, neural tube midline deficit, craniofa- cial
dysmorphology) in fetal alcohol spectrum disorder. Two such hypotheses emerge from the current
study. The first is that alcohol causes a delay in development of the nervous system by inhibiting
specific sets of genes involved in neural development (Ngn, Nhlh, Sox, Igf, Ntf, and Egf). The second is
that neural tube defects are mediated by the inhibition of genes in the epidermal growth factor
signaling pathway and genes encoding histone variants.

Fetal Alcohol Spectrum Disorders: The Epigenetic Perspective - Philip C Haycock
Ethanol is a classic teratogen capable of inducing a wide range of developmental abnormalities. Studies in
animal models suggest that differences in timing and dosage underlie this variability, with three particularly
important developmental periods: preconception, preimplantation and gastrulation. These periods of
teratogenesis correlate with peak periods of epigenetic reprogramming which, together with the evidence
that ethanol interferes with one carbon metabolism, DNA-methylation, histone modifications and noncoding
RNA (ncRNA), suggests an important role for epigenetic mechanisms in the aetiology of fetal alcohol
spectrum disorders (FASD). In addition to a number of testable hypotheses, an epigenetic model suggests
that the concept of a ‘‘fetal alcohol spectrum’’ should be expanded to include ‘‘preconceptional
effects’’. This proposal has important public health implications, highlighting the urgency of research into
the epigenetic basis of FASD.
Mechanisms of Alcohol-Induced Damage to the Developing Nervous System Charles R.
Goodlett, Ph. D. , and Kristin H. Horn
Recent lterature reviews confirm that no single putative mechanism can account for all the
components and variations of the anatomical and behavioral characteristics ( i. e. , phenotypes)
found in children prenatally exposed to alcohol ( Abel and Hannigan 1995; Guerri 1998; Maier et
al. 1996; Michaelis 1990; Michaelis and Michaelis 1994; Phillips et al. 1989; Schenker et al. 1990;
West et al. 1994)
Some of alcohol s harmful effects on the developing organism probably result from alcohol-
induced changes in gene expression the processes through which the genetic information
encoded in the genes is converted into gene products ( i. e. , proteins) . Because a large number
of genes are active at any given time in a cell, analyses of alcohol-induced changes were
previously considered both difficult and time consuming. However, new genomic technologies,
such as gene microchip arrays, provide a means to screen the expression of large numbers of
genes and identify alcohol-induced changes. This approach allows researchers to evaluate
simultaneously changes in the expression of thousands of genes, thereby providing snapshots of
alcohol-induced increases or decreases in gene expression. This technique may reveal otherwise
undetected patterns of changes across many groups of related genes ( i. e. , gene families) .
Slide - 23
Neurotransmitters, Neuromodulator Systems and Hormones
From Wikipedia - Acetylcholine (often abbreviated ACh) acts as a neurotransmitter in both the
peripheral nervous system (PNS) and central nervous system (CNS)
Acetylcholine is one of many neurotransmitters in the autonomic nervous system (ANS) and is the
only neurotransmitter used in the motor division of the somatic nervous system (sensory neurons
use glutamate and various peptides at their synapses). Acetylcholine is also the principal
neurotransmitter in all autonomic ganglia. In cardiac tissue acetylcholine neurotransmission has an
inhibitory effect, which lowers heart rate. However, acetylcholine also behaves as an excitatory
neurotransmitter at neuromuscular junctions in skeletal muscle. In the central nervous system,
acetylcholine and the associated neurons form a neurotransmitter system, the cholinergic system,
which tends to cause anti-excitatory actions. In the central nervous system, ACh has a variety of
effects as a neuromodulator upon plasticity, arousal and reward. ACh has an important role in the
enhancement of sensory perceptions when we wake up and in sustaining attention. ACh has also
been shown to promote REM sleep.
Some forms of learning and plasticity in cortex appear dependent on the presence of acetylcholine.

Dopamine (abbreviated as DA) plays a major role in the brain system that is responsible for reward-
driven learning. a variety of highly addictive drugs, including stimulants such as cocaine and
methamphetamine, act directly on the dopamine system. Personality traits such as extraversion and
reward seeking have been linked to higher sensitivity to rewarding stimuli of the mesolimbic
dopamine system.
Dopamine has many functions in the brain, including important roles in behavior and cognition,
voluntary movement, motivation, punishment and reward, inhibition of prolactin production
(involved in lactation and sexual gratification), sleep, dreaming, mood, attention, working memory,
and learning. Dopaminergic neurons (i.e., neurons whose primary neurotransmitter is dopamine) are
present chiefly in the ventral tegmental area (VTA) of the midbrain, the substantia nigra pars
compacta, and the arcuate nucleus of the hypothalamus.
Dopamine reduces the influence of the indirect pathway while increasing the actions of the direct
pathway within the basal ganglia. Insufficient dopamine biosynthesis in the dopaminergic neurons
can cause Parkinson's disease, a condition in which one loses the ability to execute smooth,
controlled movements.
Dopamine is the primary neuroendocrine inhibitor of the secretion of prolactin from the anterior
pituitary gland.
In the frontal lobes, dopamine controls the flow of information from other areas of the brain.
Dopamine disorders in this region of the brain can cause a decline in neurocognitive functions,
especially memory, attention, and problem-solving. Reduced dopamine concentrations in the
prefrontal cortex are thought to contribute to attention deficit disorder.
γ-Aminobutyric - GABA is the chief inhibitory neurotransmitter in the mammalian central
nervous system. It plays a role in regulating neuronal excitability throughout the nervous
system. In humans, GABA is also directly responsible for the regulation of muscle tone.
GABA acts at inhibitory synapses in the brain by binding to specific transmembrane
receptors in the plasma membrane of both pre- and postsynaptic neuronal processes. This
binding causes the opening of ion channels to allow the flow of either negatively charged
chloride ions into the cell or positively charged potassium ions out of the cell. This action
results in a negative change in the transmembrane potential, usually causing
hyperpolarization.
While GABA is an inhibitory transmitter in the mature brain, its actions are primarily
excitatory in the developing brain.
GABA regulates the proliferation of neural progenitor cells the migration and differentiation[
the elongation of neurites[ and the formation of synapses.
GABA also regulates the growth of embryonic and neural stem cells. GABA can inﬂuence
the development of neural progenitor cells via brain-derived neurotrophic factor (BDNF)
expression.[24] GABA activates the GABAA receptor, causing cell cycle arrest in the S-
phase, limiting growth.
Publications
Effects of ethanol, given during pregnancy, on the offspring dopaminergic system
Laura Lucchi, Vito Covelli2, Vesselin V. Petkov3, Pier-Franco Spano, Marco Trabucchi [1983]

Our work demonstrates long-lasting modifications of dopaminergic neuronal function after
exposure of the experimental animal to ethanol during fetal life. In particular, a decreased
receptor function has been observed in rats exposed to ethanol only during the perinatal
period. In the same group of rats, diminished receptor activity leads to an enhancement in
DOPAC content still detectable after a long period from cessation of ethanol treatment.
Neurochemical data are reinforced by behavioral observations. In fact, a significant
decrease of spontaneous locomotor activity in the rats chronically treated with ethanol
during fetal life was observed. In addition, the altered response of locomotor activity after
drug administration may be ascribed to the modified dopaminergic function. With this
experimental approach we assume that the action of ethanol on the central nervous
system may be a marker of its teratogenic effect.
FOCUS ON: NEUROTRANSMITTER SYSTEMS
C. Fernando Valenzuela, M.D., Ph.D.; Michael P. Puglia; and Stefano Zucca, M.Sc.
These findings suggest that daily consumption of small amounts of ethanol (such as a glass of wine
with meals) during the first and second trimesters of pregnancy could have significant effects on the
development of GABAergic neurons in the fetus. Given the prominent role of GABA during
development, this could significantly affect the normal development of cortical neuronal circuits.
Collectively, these studies emphasize that ethanol can affect the function of the GABA
neurotransmitter system even before synapses have been formed and that neurochemical imbalances
can have profound consequences on early neuronal development.
Potent effects of ethanol exposure on GABA transmission during the third trimester–equivalent
period also havebeendocumentedinthehippocampus—another brain region that is important for
learning and memory processes.
Impact of Ethanol on the Developing GABAergic System
RICARDO NOBORO ISAYAMA,1 PAULO EMILIO CORREA LEITE,2 JEAN PIERRE MENDES LIMA,1,3
DANIELA UZIEL,1*
AND EDNA NANAMI YAMASAKI3,4*
FAS and FASD are multifactorial entities with functional and structural abnormalities affecting not
only the nervous system but also the whole organism. As such, we cannot conclusively establish which
of the different factors (neurotransmitters, adhesion molecules, cell death, transcription factors,
trophic factors, etc.) reported in the literature are the major players in the syndrome. More likely, the
final result is a complex interaction, with some of the players being up or down- regulated to
compensate for the loss of function. These compensatory changes can be maladaptative, resulting in
FAS or FASD.
We present here a review on the effects of ethanol on the GABAergic system. GABA is the major
inhibitory neurotransmitter in the nervous system, with the peculiarity of being an excitatory
molecule during early stages of development. GABA, as with other neurotransmitters, also has
nonsynaptic functions, being involved in several developmental steps such as proliferation, migration,
differentiation, and synaptogenesis. The GABAergic system seems to respond to the ethanol insult
with compensatory changes, but we do not know if the compensatory mechanisms in place allow for
normal function. An imbalance in the precise anatomical and functional inhibitory connectivity will
lead to physiological and cognitive deficits that could contribute to the findings in FAS.

Serotonin - 5- hydroxytryptamine (5-HT) is a monoamine neurotransmitter. Biochemically
derived from tryptophan, serotonin is primarily found in the gastrointestinal (GI) tract, platelets, and
in the central nervous system (CNS) of animals including humans. It is popularly thought to be a
contributor to feelings of well-being and happiness.
Approximately 90% of the human body's total serotonin is located in the enterochromaffin cells in
the alimentary canal (gut), where it is used to regulate intestinal movements. The remainder is
synthesized in serotonergic neurons of the CNS, where it has various functions. These include the
regulation of mood, appetite, and sleep. Serotonin also has some cognitive functions, including
memory and learning. Modulation of serotonin at synapses is thought to be a major action of several
classes of pharmacological antidepressants. Serotonin decreases appetite. Serotonin is known to
regulate aging, learning and memory.
Slide - 24
From Wikipedia - Neurotrophins are a family of proteins that induce the survival, development, and
function of neurons.
They belong to a class of growth factors, secreted proteins that are capable of signaling particular
cells to survive, differentiate, or grow. Growth factors such as neurotrophins that promote the
survival of neurons are known as neurotrophic factors. Neurotrophic factors are secreted by target
tissue and act by preventing the associated neuron from initiating programmed cell death - thus
allowing the neurons to survive. Neurotrophins also induce differentiation of progenitor cells, to
form neurons.
Although the vast majority of neurons in the mammalian brain are formed prenatally, parts of the
adult brain (for example, the hippocampus) retain the ability to grow new neurons from neural stem
cells a process known as neurogenesis. Neurotrophins are chemicals that help to stimulate and
control neurogenesis.
During the development of the vertebrate nervous system, many neurons become redundant
(because they have died, failed to connect to target cells, etc.) and are eliminated. At the same time,
developing neurons send out axon outgrowths that contact their target cells. Such cells control their
degree of innervation (the number of axon connections) by the secretion of various specific
neurotrophic factors that are essential for neuron survival.
Epigenetic regulation of brain-derived neurotrophic factor: implications in
neurodevelopment and behavior.
Schanker BD.
Several recent research findings have implicated brain-derived neurotrophic factor (BDNF) as a
mediator of neuronal plasticity. The BDNF gene is under extensive epigenetic regulation, which
modulates how much or how little environmental experiences become encoded within neurons and
neural circuits.
Epigenetic regulation of reelin and brain-derived neurotrophic factor genes in long-term
potentiation in rat medial prefrontal cortex.
Sui L, Wang Y, Ju LH, Chen M.
These findings suggest that epigenetic regulation of DNA demethylation and histone acetylation of
target genes, such as reelin and bdnf, might underlie the mechanisms of synaptic plasticity and
memory retention in the mPFC.

Hippocampal cell loss and neurogenesis after fetal alcohol exposure:
insights from different rodent models.
Gil-Mohapel J, Boehme F, Kainer L, Christie BR.
Prenatal ethanol exposure is invariably detrimental to the developing central nervous
system and the hippocampus is particularly sensitive to the teratogenic effects of ethanol.
Prenatal ethanol exposure has been shown to result in hippocampal cell loss, altered
neuronal morphology and impaired performance on hippocampal-dependent learning and
memory tasks in rodents. The dentate gyrus (DG) of the hippocampus is one of the few
brain regions where neurogenesis continues into adulthood. This process appears to have
functional significance and these newly generated neurons are believed to play important
functions in learning and memory. Recently, several groups have shown that adult
hippocampal neurogenesis is compromised in animal models of fetal alcohol spectrum
disorders (FASD). The direction and magnitude of any changes in neurogenesis, however,
appear to depend on a variety of factors that include: the rodent model used; the blood
alcohol concentration achieved; the developmental time point when alcohol was
administered; and the frequency of ethanol exposure.
Slide – 25
When all the evidence is examined we have to ask what relationship PAE has to these
conditions?
Attention, Mood and Personality Disorders
Addictions
Cancer
Infectious Diseases
We know since 1965 that 95% of those diagnosed FASD will have diagnoses from the
DSM, often multiple.
We have seen how those afflicted with FASD are at risk for infections, cancer and
addictions as a result of the PAE and Epigenetics
Slide – 26
The ubiquitous effects of PAE on the developing fetus have for many years been puzzling
and even a source of skepticism. Ten years ago there was little discussion about
epigentics and changes of gene expression. Today there is an epigenetic industry being
developed, searching for the cure of all our illnesses.
For the foreseeable future only the few who are knowledgeable and concerned will be the
ones advocating for epigenetic research to be applied to FASD and PAE.
Publications
FOCUS ON: Epigenetics And Fetal Alcohol Spectrum Disorders -Michael S. Kobor,
Ph.D., and Joanne Weinberg, Ph.D. National Institute on Alcohol Abuse and
Alcoholism
Perhaps the best understood epigenetic mechanism involves the regulation of gene

expression—that is, the process through which the genetic information encoded in
the DNA directs the production of RNA and proteins. In general, epigenetic
modifications serve to establish and maintain different gene expression programs
in different cell types. Differential gene expression ensures that the more than 200
different cell types that make up mammalian tissues differ in phenotype or
appearance, even though almost all cells in an organism essentially share the
same genetic information. Epigenetic mechanisms contribute, at least in part, to
this differential gene expression. Investigation of the role of epigenetic mechanisms
in the adverse effects of prenatal alcohol exposure in both humans and animal
models still is at an early stage, although its potential role is becoming more widely
recognized (Haycock 2009).
However, once our child is an adult then he/she will benefit from the research into [the
resulting] adult conditions

Epigenetic regulation in alcoholic liver disease - Natalia A Osna, MD, PhD,
The interplay of epigenetic mechanisms and their influence on gene transcription in ALD is
evolving.
Epigenetic alterations associated with acute and chronic alcohol exposure of hepatocytes and
immune cells in relation to ALD is discussed in this review. Studies thus far have shown that
alcohol exposure, probably via oxidative stress, exhibits differential regulation of acetylation,
phosphory- lation and methylation of histones that regulate chromatin remodeling and gene
expression. The effects of alcohol on DNA methylation in hepatocytes and miRNA regulation
have been elucidated. An integrative approach of the various mechanisms that lead to genomic
imprinting during alcohol exposure will identify novel pathways in the alcoholic liver and
support epigenetic therapeutic interventions.
Slide – 28
Publications
Do we deal with it now >
Ethanol Exposure Affects Glial Fibrillary Acidic Protein Gene Expression and
Transcription During Rat Brain Development
S. Vallés, J. Pitarch, ~J. Renau-Piqueras, and C. Guerri
Glial cells are one of the most abundant cell types in the vertebrate CNS, and they are
essential for normal development of the nervous system. They provide the structure and
nutritive support for developing neurons and are involved in many functions including
stimulation of neurite out- growth and guidance of migrating neurons, synaptogenesis and
synaptic plasticity, regulation of ions and transmitters in the microenvironment, production
of several growth factors, and regulation of water, energy. and nutrient support of neurons.

Therefore, the present data support the hypothesis that ethanol-induced GFAP gene
expression and astroglial damage could be a potentially important mechanism involved in the
neurological and neurobehavioral dysfunctions associated with fetal alcohol syndrome and
with prenatal alcohol exposure.
Or wait until later >
The effect of chronic alcoholism on epigenetic patterns regulating gene
expression and neurodegeneration in the
human brain - SOFIA JOHANSSON
Chronic alcoholism causes about 3.2% of all deaths worldwide. It has an
association to about 60 different diseases and injuries with enormous
social and economical impact.
Slide – 29
Preconception, Preimplantation, Gastrulation.
From Wikipedia - Gastrulation is a phase early in the embryonic development of most
animals, during which the single-layered blastula is reorganized into a trilaminar ("three-
layered") structure known as the gastrula. These three germ layers are known as the
ectoderm, mesoderm, and endoderm. Each layer gives rise to specific tissues andorgans in
the developing embryo. The ectoderm gives rise toepidermis, and to the neural crest and
other tissues that will later form the nervous system. The mesoderm is found between the
ectoderm and the endoderm and gives rise to somites, which form muscle; the cartilage of the
ribs and vertebrae; the dermis, the notochord, blood and blood vessels, bone, and connective
tissue. The endoderm gives rise to the epithelium of the digestive system and respiratory
system, and organs associated with the digestive system, such as the liver and pancreas.
Publications
FOCUS ON: EPIGENETICS AND FETAL ALCOHOL SPECTRUM DISORDERS
Michael S. Kobor, Ph.D., and Joanne Weinberg, Ph.D.
Preconception>
More recent studies support these findings of preconception effects mediated by
paternal alcohol consumption. For example, reduced birth weight and length, reduced
litter size, and increased malformations in the offspring have been reported.
Transmission of these epigenetic changes to the offspring through fertilization possibly
could alter gene expression in the fetus, thus affecting prenatal development.
Preconception effects of maternal alcohol consumption also have been reported,
including effects on birth weight and growth.

Preimplantation>
The time after the egg cell has been fertilized and before it implants in the uterus—
encompasses the first 4 to 6 days of mouse and rat development, which corresponds
roughly to the first 2 weeks of human pregnancy. The investigators found that although
placentas and embryos of alcohol-exposed mothers showed severe growth retardation
compared with those of control animals, alcohol exposure did not alter overall DNA
methylation in the embryos.
Gastrulation >
Gastrulation is the stage early during embryonic development that follows implantation
of the embryo into the uterine wall, which in mice is completed by day 6. Gastrulation
encompasses the onset of organ development, which occurs on days 7 to 14 of gestation
in the mouse and during weeks 3 to 8 in humans. During this period, extensive cellular
differentiation occurs and the various organs begin to form. As mentioned above, the
gastrulation period generally is considered the most sensitive to teratogenic insult
because differentiating cells appear to be particularly vulnerable to the teratogenic
effects of alcohol. Further support for the role of epigenetic mechanisms during
gastrulation comes from a recent study by Liu and colleagues (2009), who cultured
whole mouse embryos and exposed them to a high dose of alcohol3 for 44 hours,
beginning at embryonic day 8.25. Overall, the investigators observed delayed and
reduced growth, as well as numerous abnormalities, including alterations in the heart,
the caudal neural tube (i.e., tail end of the developing central nervous system), brain
vesicles, optic system, and limb buds.

Role of Epigenetics in Developmental Biology and Transgenerational Inheritance
Michael K. Skinner*
The inheritance of environmen- tally induced phenotypes is the origin of the concept of
epigenetics . In the event these environmental factors modify the epigenome of the
germ line and this becomes permanently programmed (imprinted) then the altered
epigenome and phenotype become transgenerational and appear in subsequent
progeny and generations in the absence of any further environmental exposures . The
mechanism involves the actions of an environmental factor at a critical time of gonadal
sex determination in the mammalian fetus when the germ line cell fate is determined
and the primordial germ cell differentiates into a male or female germ lineage . During
this critical period of development of the germ line an erasure of DNA methylation
occurs and then upon gonadal sex determination the germ line DNA is remethylated in
a male or female specific manner.

Slide – 30
It is suggested that the process of epigenetic changes of gene expression was designed
to protect the fetus from negative, although perhaps natural, environmental insults, but

that the changes resulted in negative in later years as an adult. Also, the process makes
the gene [expression] vulnerable to agents such as alcohol, not a natural environmental
insult.

Publications
Epigenetics and fetal adaptation to perinatal events: Diversity through fidelity -
L. A. Joss-Moore, D. B. Metcalfe, K. H. Albertine, R. A. McKnight, and R. H. Lane
The ability to adapt to the in utero environment provides a means of increasing the
odds of fetal survival under a variety of conditions. However, this short-term gain
appears to be accompanied by a long-term cost, and it is now well established that fetal
insults are associated with an increased susceptibility to several adult- onset metabolic
disorders, including cardiovascular disease, insulin resistance, and obesity . Insults
generally tend to involve an interruption of nutrients to the fetus, with subsequent in
utero growth restriction. Common insults include fetal undernutrition, hypoxia, or both,
resulting from maternal malnutrition or utero- placental insufficiency.
Another important concept is that traditional epigenetics tends to involve gene
expression in terms of “on” or “off”; that is, genes are either silent or activated. In
contrast, epigenetic mechanisms regulating perinatal adaptation tend to be associated
with an adjustment in the expression of genes that are already being transcribed,
somewhat akin to a rheostat or volume effect.

Slide - 31
From Wikipedia - A genetic marker is a gene or DNA sequence with a known location on a
chromosome that can be used to identify individuals or species. It can be described as a
variation (which may arise due to mutation or alteration in the genomic loci) that can be
observed. A genetic marker may be a short DNA sequence, such as a sequence
surrounding a single base-pair change (single nucleotide polymorphism, SNP), or a long
one, like minisatellites.
Epigenetics and fetal adaptation to perinatal events: Diversity through fidelity -
L. A. Joss-Moore, D. B. Metcalfe, K. H. Albertine, R. A. McKnight, and R. H. Lane
We have also focused on genes that are likely to be particularly important in the context
of perinatal adaptation. Several genes have emerged as important candidate genes, one
of which is IGF-1. In multiple models of perinatal disease, IGF-1 displays epigenetic
modifications that are common, critical, and correlated with IGF-1 expression. These
are likely to be mechanistically important in the epigenetic regulation of perinatal
adaptability

Slide - 32
A possible definition of a complex system
Acknowledged features of a complex system are the following:
-- the system is composed of a large number of elements;
-- the elements are often of different types and have an essential internal
structure;
-- the elements are related by nonlinear interactions, often of several
different types;
-- the system experiences inputs at several scales.
But I argue that the main hallmark of complex systems is circular
causality, namely, the presence of feedbacks of (macroscopic) collective
properties and emergent features on the behavior of (microscopic)
elements. Elements collectively modify the surroundings, which in turn
exerts constraints on them and endow them with different possible states
or behaviors. In complex systems, knowing the features and behavior of
the single components in isolation is not sufficient to predict the behavior
of the system as a whole. Annick LESNE (CNRS UMR 7600 & IHÉS)
From Wikipedia - In chaos theory, the butterfly effect is the sensitive dependence on initial
conditions, where a small change at one place in a deterministic nonlinear system can result
in large differences to a later state . In the social sciences, unintended consequences
(sometimes unanticipated consequences or unforeseen consequences) are outcomes that
are not the ones intended by a purposeful action. Unintended consequences can be roughly
grouped into three types:
A positive, unexpected benefit (usually referred to as luck, serendipity or a windfall).
A negative, unexpected detriment occurring in addition to the desired effect of the policy
(e.g., while irrigation schemes provide people with water for agriculture, they can increase
waterborne diseases that have devastating health effects, such as schistosomiasis).
A perverse effect contrary to what was originally intended (when an intended solution makes
a problem worse)
Publication
The Law of Unintended Consequences
by Alex Tabarrok on January 24, 2008 at 7:47 am in Economics | Permalink
The law of unintended consequences is what happens when a simple system tries
to regulate a complex system

Aboriginal FASD and CNP Workshop Toronto, 6th. March, 2013

Aboriginal FASD and CNP Workshop Toronto, 6th. March, 2013

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Aboriginal FASD and CNP Workshop Toronto, 6th. March, 2013