William

Meng

During my senior year of high school, I conducted a guided research project with Mr.
Simon Holdaway, Mr. Neil Chaudhary, and a couple other students on oxidative
stress in chick forebrain cells. The research was never finished because Mr. Simon
Holdaway left the school in the middle of the project. However, I learned a great deal
in the process and would like to provide a brief summary of what we had done.

The specific question we were trying to answer is, "What chemical will offer
neuroprotective effects against oxidative stress induced by hydrogen peroxide in
chick forebrain neurons?" In the process, we wanted to determine a specific, and
validated methodology, to look for answers to that question. In 2015, the previous
group discovered that Hesperidin, an antioxidant found in orange peels, has
neuroprotective traits1. In 2016, we aimed to find another chemical that offers
similar neuroprotective properties. We spent this fall learning cell culture
techniques to grow and harvest neurons in a petri dish without contamination by
bacteria or yeast. We researched, with the help of Pubmed and other databases, for
chemicals that may have neuroprotective properties. One of our top candidates
included myricetin that has proven to reduce oxidative stress induced by H2O2 in
mice lung cells and yeast cells. To determine the amount of cell death in each
experiment, we compared the amount of LDH released through apoptosis with
hydrogen peroxide, the chemical we found, and chick forebrain cells with a control
that only contains hydrogen peroxide and chick forebrain cells. We can measure the
amount of LDH released by using a LDH assay called CytoTox 96 Non-Radioactive
Cytotoxicity Assay. By comparing the absorbance values of the resulting assay, we
can determine whether or not the chemical was effective in protecting chick
forebrain cells from neurodegeneration caused by hydrogen peroxide.

All in all, throughout the process, I learned the techniques of dissecting chicken eggs
to harvest chick forebrain neurons that can continue to divide, performing the
Western Blott protein detection, using the Bradford protein assay, and growing and
harvesting neurons in a controlled environment.

1 http://www.loomischaffee.org/uploaded/News_Images/Spring_2015/Guided_research_project.JPG