Phang Laohavirojana 1109 !

Level methylation of genetic factors and environmental factors on

schizophrenia

I. Background Information

Schizophrenia is one of the most chronic brain illness and metal disorder that should

concern. It effects peoples thinking, emotion, behaviour and perception of this word or people

surrounding. Its not a common disease comparing to other metal disorder but its not a rare

disease either. Studies shown that out of one hundred people, the probability that people will have

schizophrenia is one person and both of males and female having chance of getting schizophrenia

but males has slightly more chance developed this disease. People can get schizophrenia in ages

of 16 to 30 years old. For the symptoms of Schizophrenia is vary and variety such as difficult to

manage emotions, difficult to distinguish events that is real or unreal or struggle concentration.

However, schizophrenias symptom can be classified in three categories.

Positive symptoms, which refers to struggle in mind including hallucination, delusion,

movement disorder and non functional thought. Schizophrenias patient with positive symptoms

will hear someone speak and that voice doesnt actually exist or feeling insecure that

someones going to harm them.

Negative symptoms, which refers to abnormal of expression of emotion or behaviours included

Flat affect. Flat effect means people with this type of symptoms will reduce facial expression

or voice tone. Sometimes, people can be unhappy within reasons in everyday life.

Cognitive symptoms, refer to changing in memory or perspective of thinking, including difficult

to concentrate or pay attention on something or having a trouble on using information

immediately after learning it.

This case disadvantages to people with schizophrenia. It makes people who have schizophrenia

has difficulties in living, more likely to commit suicide and develop violences behaviour.
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Cause of Schizophrenia: Genetic factors Vs. Environment factors

The causes of schizophrenia is not fully unknown. However, there are several risk factor that can

cause schizophrenia and this all about nature versus nurture. For Nature, It is associated to

genetic factors. Many of researches shown that Schizophrenia runs in relative and family. Based

on the British Journal of Psychiatry about Genes, environment and schizophrenia, they did an

experiment on identical twins and adopted child who has parents or families that have

schizophrenia. The result showing that the identical twins have about 50 percent developing

schizophrenia. For adopted children that their parents has the disease, they also having a

schizophrenia too. This showing that Schizophrenia is partly of heredity and genes. However,

people who have schizophrenia not have family or relatives have this disease but they are affected

by the environment factors. Scientists think that some of the environment factors is partly make the

genes changing and as the result develop to disease. For schizophrenia, the environment factors

that might involve in is lack of nutrition during pregnancy, sexual abuse in childhood, anxiety,

neighbourhood and etc. These non-magnetic factors is partly of altered genes and epigenetic

marker.

DNA methylation, Schizophrenia and BDNF genes

DNA methylation is an epigenetic marks that involve in human development and brain

development too. To make Based on the Nature Neuroscience, they suggested that DNA

methylation which is an epigenetic markers which addition of CH3 group or methyl group can affect

gene sequencing influences the brain development whether it abnormal or not and it results people

have higher risk of schizophrenia. And for schizophrenia, there is no single specific genes that

cause this disease. However, in this experiment, we going to test the level of DNA methylation from

BDNF genes. BDNF(Brain-derived neurotrophic factor) genes is a protein that associated to

encode in human nerve growth. BSNF genes also plays a part of schizophrenia and this disease is

related to brain development and plasticity (the ability of brain that can modify its shape and

structure by the changes of the environment.).If we consider the role of BSFN which are synapses

regulation, brain plasticity regulation, learning, memory and social interaction then you can see that
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It related and if the BSNF genes is altered, it can say that it can cause the brain development

changes or synaptic of the brain can disconnect. This can be found in the brain of people with

schizophrenia.

II. Objectives

A. Define the which factors; genetic and environment can cause the schizophrenia.

B. Define the exactly cause of schizophrenia.

C. Can apply to the new treatment in the future.

Independent variables: 1. Environment factors 2. Genetic factors (Family history)

Dependent variables: Level of DNA methylation

Control variables: 1. Ages 2.Females 3. Ethic origin 4. Geographical area

III. Question

What factors that has a huge effect on getting schizophrenia between genetics

factor or environmental factors ?

IV. Hypothesis

If the we tested the level of DNA methylation then the environmental factors will

have more probability to increase and altered DNA methylation in BSFN genes.

V. Materials

1. Group of one hundred females human

Can be classified by diagram below:

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Environmental
factors:
sexual abuse in
Family childhood
Background:
with
schizophrenia

Family No
Background: environment
with factors
Females schizophrenia

100 (Age: 20-30

people years old)

Environmental
Family factors:
Background sexual abuse in
without SZ childhood

No
no family
environment
history of
factors
schizophrenia

2. Clotted blood sample

3. Proteinase K digestion

4. Salt (6N Saturated NaCl)

6. 90% ethanol

7. Methylation-specific PCR

8. Primers

9. UV light

10. Ethidium Bromide

11. polypropylene tube

12. Lysis buffer

13. Benchmark BlotBoy 3D Low Speed Rockers

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14. Centrifuge

15. Luminometric Methylation Assay (LUMA)

VI. Procedures

1. Take clotted blood sample of all the group of people and put it in polypropylene tube.

2. Extracted BDNF genes from blood clotted samples by using the salt-out method.

Bring the blood sample that contained in polypropylene tube.

If you want to isolate DNA the same day that you do a salt-chloroform method or if

you want to use in future, keep it at - 20 celsius.

Add a lysis buffer to the blood sample.

Keep on ice for 10 to 15 minutes.

After that, centrifuge the sample at 10,000 revolution per minutes and about 4

celsius.

Put Proteinase K in the blood sample contained polypropylene tube.

Leave it overnight at 56 celsius on Benchmark BlotBoy 3D Low Speed Rockers.

Adding salt and mixed vigorously for 15 minutes.

After that use centrifuge to spin the blood sample again in 10,000 revolution per

minute in 10 minutes at 4 celsius.

Precipitated the DNA by mixing the cold 90% alcohol.

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3. Determine the DNA methylation status in extracted BFSN DNA in every group of blood

sample by using Methylation-specific PCR (MSP).

Methylation-specific PCR with two primers which can help defined that which one

is methylated or unmethylated DNA.

4. After the sample was passed the Methylation-specific PCR method, use ethidium

bromide stained the product in order to visualise the result.

5. After that illustrated the product by using UV light.

6. Use Luminometric Methylation Assay (LUMA) tools to determine the level of methylation

of each samples.

7. Record the result of each categories and Compare the level of DNA methylation

changes in each group.

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VII. Implication

People with schizophrenia can struggle in their daily life. Its difficult to people

with schizophrenia to get a job, have good work, or friends. As you can see that the

causes of schizophrenia is still fully unknown. The treatments of this disease only

help maintain the disease and focused on the eliminating the symptoms. If we can

define whether it causes from genetic factor or environmental factor more, we can

find the prevention and also the treatment that can inhibit the symptoms so

schizophrenias patient can spend their life like normal people and open opportunity

to schizophrenias patient.
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