• Epigenetic control of gene expression in

eukaryotes.
• Epigenetics involves heritable changes in gene
function, without changes to the base sequence
of DNA. These changes are caused by changes
in the environment that inhibit transcription by:
– increased methylation of the DNA or
– decreased acetylation of associated histones.
• The relevance of epigenetics on the
development and treatment of disease,
especially cancer.
 DNA - epigenetics
• Have a go at the 2 sheets to recap DNA
structure
 Epigenetics
• VERY new
• It looks at how environmental influences
alter genetic inheritance
Histones
DNA and histones
 DNA and histones
• Histones get covered in chemicals referred
to as tags.
• These tags are the epigenome
• They determine the shape of the histone-
DNA complex
• Inactive genes can
be wrapped tightly
and become
inaccessible –
ensuring they are
not read –
(epigenetic
silencing)
• Active genes are
‘looser’ and are
more easily read
and transcribed
Fixed
Epigenome - flexible
 Epigenome
• Accumulation of environmental signals
received in life – chemical tags
• From the moment of conception (or even
before)
 Dutch famine
• 1944–45
 Description/Abstract
The early embryo and periconceptional period is a window during which environmental
factors may cause permanent change in the pattern and characteristics of
development leading to risk of adult onset disease. This has now been demonstrated
across small and large animal models and also in the human. Most evidence of
periconceptional ‘programming’ has emerged from maternal nutritional models but also
other in vivo and in vitro conditions including assisted reproductive treatments, show
consistent outcomes. This short review first reports on the range of environmental in
vivo and in vitro periconceptional models and resulting long-term outcomes. Second, it
uses the rodent maternal low protein diet model restricted to the preimplantation period
and considers the stepwise maternal-embryonic dialogue that comprises the induction
of programming. This dialogue leads to cellular and epigenetic responses by the
embryo, mainly identified in the extra-embryonic cell lineages, and underpins an
apparently permanent change in the growth trajectory during pregnancy and
associates with increased cardiometabolic and behavioural disease in adulthood.

University of Southampton logo

Embryos, DOHaD and David Barker

Fleming, Thomas, Velazquez, Miguel and Eckert, Judith (2015), September 2014;
Prenatal Exposures and Health Outcomes] Journal of Developmental Origins of Health
and Disease, 6, (5), 377-383. (doi:10.1017/S2040174415001105). (PMID:25952250).
Maternal undernutrition affects neurogenesis in the foetal mouse brain
Airey, Chris J., Smith, Phoebe J., Restall, Katie, Marfy-Smith, Stephanie J., Fleming, Tom P. and Willaime-Morawek, Sandrine (2015) Maternal
undernutrition affects neurogenesis in the foetal mouse brain. [in special issue: ISDN 2014 Abstracts] International Journal of Developmental
Neuroscience, 47, part A, 72. (doi:10.1016/j.ijdevneu.2015.04.197).(PMID:26531520)

In the modern world, malnutrition is widespread and the World Health Organisation states it as the greatest
threat to the world’s public health. With this in mind many studies tried to investigate the consequences of
inadequate nutrition on human development and health. Abnormal anxiety-related behaviour and increased risk
of schizophrenia have been shown following a maternal malnutrition during pregnancy. Whilst previous studies
indicate maternal low protein diet (LPD) to have a negative effect on foetal brain development, we have
investigated specifically whether maternal LPD negatively affects neural stem cell derivation and neurogenesis
in the foetal mouse brain.

Female mice were fed different diets from conception: normal protein diet (NPD), low protein diet (LPD) or
embryonic LPD (Emb-LPD: LPD until E3.5, NPD thereafter).

The number of primary neurospheres generated from E14.5 ganglionic eminences was decreased in LPD vs
NPD. No overall difference in the formation of secondary neurospheres was detected between treatments
following bulk passaging of the primary spheres. Our results also show that in both LPD and Emb-LPD groups,
cell proliferation is decreased in vitro and in vivo, and this might explain the decrease in neurosphere numbers.

Immunostaining of E14.5 brain sections revealed that in both LPD and Emb-LPD groups, the number of neural
stem cells (Sox2 positive) decreased compared to NPD. However, immature neurons (Beta-III-tubulin positive)
are increased in Emb-LPD and decreased in LPD, compared to NPD, suggesting that LPD affects
neurogenesis.

Analysis of adult offspring brains by Western-blots revealed trends toward a decrease of Beta-III-tubulin and an
increase of NeuN (mature neurons) in the cortex of LPD, compared to NPD.

Overall, our work suggests a profound effect of maternal protein diet on neural stem cells and neurogenesis in
the foetus, which may persist in the adult offspring brain.
 epigenome
• They can activate or inhibit particular sets
of genes

• Different genes are more or less sensitive

to particular signals
 The environmental signals
• They stimulate proteins to chaperone its
messages to the cell
• Signals in chaperoned through a variety of
proteins and makes it into the nucleus

• It only has two effects

 Acetlyation
• Acetylation of histones leads to activation
or inhibition
 Methylation
• Methylation of DNA by attraction of
enzymes that can add or remove methyl
groups
 The DNA histone complex
• A weak association with histone = loosely
packed DNA
• DNA is accessible by transcription factors
– mRNA can be made
• Strong association with histone = tightly
packed
• Not accessible by transcription factors
 Decreased acetlyation
• Acetylation = acetyl group is transferred to
a molecule
• Group donating is acetylcoenzyme A
• Deacetlyation = acetyl group is removed
 Decreased acetlyation
Increases the positive charges on histones
and therefore increases their attraction to
the negative phosphate groups.

Association between histone and DNA is

stronger and therefore no transcription

i.e. gene is switched off

 Increased methylation
• Methylation = addition of methyl group to
molecules
• It is added to cytosine bases
• Normally is inhibitory

• 1) prevents binding of transcription factors

• 2) attracting proteins that condense the
DNA – histone complex (by inducing
deactylation if the histones). Therefore is
inaccessible to transcription factors.
 Why do we say that DNA is a fixed
Epigenetics
What is epigenetics?
code?

Why do we say that the epigenome is

flexible?
What is the epigenome?

What kind of environmental signals

What are histones and what do they do? affect the epigenome?

How do they affect the epigenome?

Label this diagram

What affects can these signals have?

How does it occur?
 Table – inaccessible
Histones

DNA

DNA-histone complex

Chromatin type

Transcription factors

Genes
 Table – accessible
Histones

DNA

DNA-histone complex

Chromatin type

Transcription factors

Genes
 Task
• Draw 5 diagrams
1.Normal histone structure
2.Acetlyation
3.Deacetylation
4.Methylation
5.Demethylation
• Epigenetic control of gene expression in
eukaryotes.
• Epigenetics involves heritable changes in gene
function, without changes to the base sequence
of DNA. These changes are caused by changes
in the environment that inhibit transcription by:
– increased methylation of the DNA or
– decreased acetylation of associated histones.
• The relevance of epigenetics on the
development and treatment of disease,
especially cancer.
 Relevance
• Periconceptional issues – although
thought that they could be erased
research shows this to be either not true or
not effective
 Description/Abstract
The early embryo and periconceptional period is a window during which environmental
factors may cause permanent change in the pattern and characteristics of
development leading to risk of adult onset disease. This has now been demonstrated
across small and large animal models and also in the human. Most evidence of
periconceptional ‘programming’ has emerged from maternal nutritional models but also
other in vivo and in vitro conditions including assisted reproductive treatments, show
consistent outcomes. This short review first reports on the range of environmental in
vivo and in vitro periconceptional models and resulting long-term outcomes. Second, it
uses the rodent maternal low protein diet model restricted to the preimplantation period
and considers the stepwise maternal-embryonic dialogue that comprises the induction
of programming. This dialogue leads to cellular and epigenetic responses by the
embryo, mainly identified in the extra-embryonic cell lineages, and underpins an
apparently permanent change in the growth trajectory during pregnancy and
associates with increased cardiometabolic and behavioural disease in adulthood.

University of Southampton logo

Embryos, DOHaD and David Barker

Fleming, Thomas, Velazquez, Miguel and Eckert, Judith (2015), September 2014;
Prenatal Exposures and Health Outcomes] Journal of Developmental Origins of Health
and Disease, 6, (5), 377-383. (doi:10.1017/S2040174415001105). (PMID:25952250).
• Foetal issues (gestational diabetes)
Maternal undernutrition affects neurogenesis in the foetal mouse brain
Airey, Chris J., Smith, Phoebe J., Restall, Katie, Marfy-Smith, Stephanie J., Fleming, Tom P. and Willaime-Morawek, Sandrine (2015) Maternal
undernutrition affects neurogenesis in the foetal mouse brain. [in special issue: ISDN 2014 Abstracts] International Journal of Developmental
Neuroscience, 47, part A, 72. (doi:10.1016/j.ijdevneu.2015.04.197).(PMID:26531520)

In the modern world, malnutrition is widespread and the World Health Organisation states it as the greatest
threat to the world’s public health. With this in mind many studies tried to investigate the consequences of
inadequate nutrition on human development and health. Abnormal anxiety-related behaviour and increased risk
of schizophrenia have been shown following a maternal malnutrition during pregnancy. Whilst previous studies
indicate maternal low protein diet (LPD) to have a negative effect on foetal brain development, we have
investigated specifically whether maternal LPD negatively affects neural stem cell derivation and neurogenesis
in the foetal mouse brain.

Female mice were fed different diets from conception: normal protein diet (NPD), low protein diet (LPD) or
embryonic LPD (Emb-LPD: LPD until E3.5, NPD thereafter).

The number of primary neurospheres generated from E14.5 ganglionic eminences was decreased in LPD vs
NPD. No overall difference in the formation of secondary neurospheres was detected between treatments
following bulk passaging of the primary spheres. Our results also show that in both LPD and Emb-LPD groups,
cell proliferation is decreased in vitro and in vivo, and this might explain the decrease in neurosphere numbers.

Immunostaining of E14.5 brain sections revealed that in both LPD and Emb-LPD groups, the number of neural
stem cells (Sox2 positive) decreased compared to NPD. However, immature neurons (Beta-III-tubulin positive)
are increased in Emb-LPD and decreased in LPD, compared to NPD, suggesting that LPD affects
neurogenesis.

Analysis of adult offspring brains by Western-blots revealed trends toward a decrease of Beta-III-tubulin and an
increase of NeuN (mature neurons) in the cortex of LPD, compared to NPD.

Overall, our work suggests a profound effect of maternal protein diet on neural stem cells and neurogenesis in
the foetus, which may persist in the adult offspring brain.
 Cancer
• Incorrect activation of a normally inactive
gene linked with cancer
• Patients with colorectal cancer had less
DNA methylation that normal = higher than
normal gene activity (more genes turned
on)
 Cancer
• DNA in promoter regions have no
methylation normally.
• In cancer they can become highly
methylated causing genes to be “switched
off”
• This is seen in patients who develop
cancer
 Cancer
• In inherited cancer, increased methylation
of those specific genes has led to some
“protective” genes being switched off

• Can therefore lead to cancer

 Treating diseases
• Drugs to inhibit enzymes involved in
acetylation or methylation
• Inhibit enzymes that cause methylation =
reactivating silenced genes
• MUST be specific to the cancer genes
though – could cause cancer elsewhere
 Diagnosing disease
• Development of diagnostics to tests early
for cancer, brain disorders and arthritis
• Identify methylation and acetylation
relative to ‘normal’ levels.
• Can look at particular genes
• =early detection=early treatment= higher
survival rate
 Task
• How do we think the epigenome
contributes to causes of disease?
• How can we utilise our knowledge of
epigenetic causes of disease to produce
treatments for them?
• How might an understanding of
epigenetics be useful for diagnosing
disease?
 P.P style questions
1. Define what is meant by epigenetics?
(2marks)
2. Oestrogen, methyl groups and acetyl
groups are control factors that can play a
role in initiating transcription. For each
one, state how it affects transcription. (3
marks)