Kayson Nguyen - Friday 10:00 Lab

Gene Expression Analysis of Zebrafish Heart Regeneration

Introduction : In Nature, injured hearts of mammals are unable to regenerate. Scarring

is what the hearts do when they are injured for mammals. In comparison with

Zebrafish, Zebrafish are able to regenerate after having heart injury and/or amputation.

The Zebrafishs heart clots up to stop bleeding. After about 2 months the heart heals

almost fully. Analyzing the mechanics behind the regeneration of Zebrafish can

possibly lead the way to therapeutic treatments for humans with heart injury. Focus

analysis on the secreted molecules from the wound and analyze them through gene

expression. Many genes were found to be in the secreted molecules so they must find

out which ones were in common between stages of 3, 7, and 14 days in the experiment.

Reasons for Research : About a million people die in the U.S. due to heart related

injury. Heart disease in particular kills 610,000 people per year in the United States.

That is 1 in 4 deaths if our country related to the heart. With the knowledge of Zebrafish

being able to regenerate their own heart, researchers have embarked on the task to find

out the mechanisms behind it. In order to possibly create methods to help with heart

treatments so that they can help prevent the deaths of thousands of people in the

future.
Method and Materials : Zebrafish were raised to become healthy adults. They were

then used for heart amputation. The Researchers took data at the points of 3 days, 7

days, and 14 days to see the progress of heart regeneration. Then they removed the

hearts to analyze. They analyzed by extracting the total amount of RNA produced in

that time. They extracted it by using a tool called TRIzol reagent. Once that was done,

they ran it through absorbances of 260 and 280. As well as running it through gel

electrophoresis to find out the amount and caliber of the RNA extracted. They then

looked at these proteins to compare them to genes.

Results : Around 662 genes were found to be expressed during the 3, 7, and 14 day

time periods. These genes ranged vastly and they were there to do various tasks.

Such as wound response factors like : Anti-inflammatory factors, growth factors, and

tissue remodeling factors. Tissue remodeling factors occurred in the later stages of

heart regeneration, around the 7 day-14 day time period. MMP2, MMP14a, and

MMP14b are the molecules that helped form and keep the structure together during the

tissue remodeling. Things like pdgf-b, pdgf-a, and granulin-a were only some of the

upregulated that were related to the growth factor. It was concluded that these growth

factors that appeared signalled for the initiation of heart regeneration. So they then

tested the increase of these molecules in Zebrafish with injured hearts. As well as

testing the decrease of those molecules.

Discussion : This study has examined the Zebrafishs genes that were involved in

heart regeneration. Their hearts are able to regenerate due to cardiomyocyte

proliferation. How it is able to know that it needs to regenerate would be signals found

in the secreted molecules of the wound itself. Experiments of lowering and increasing

the PDGF amounts showed clear differences.

Conclusion : It was found out that in the end, the Zebrafish with the PDGF-B and

PDGF-A treatments had a upregulated increase of heart regeneration. While lowered

amounts of it decreased heart regeneration. This means that they play a significant role

in the heart regeneration of Zebrafish.

Limitations of the Study : Due to the vast amount of genes that were found to take

part in the regeneration of Zebrafish hearts, more study must be taken for each of these

genes. It was only found out that PDGF-B and PDGF-A helped with the heart

regeneration. This is only a discovery that helped Zebrafish who already have the

mechanisms for heart regeneration. This does not help solve the mystery of how to get

mammal hearts such as us humans to regenerate.

Bibliography :

http://journals.plos.org/plosbiology/article?id=10.1371/journal.pbio.0040260#s2