Determining Gene Regulation of Microglia in Glaucoma

Aurora working with micropipette tool

Students

2021
Engineering and Applied Sciences

Faculty

Project Summary

The Penn Undergraduate Research Mentorship award afforded me the opportunity to work in Dr. Claire Mitchell’s laboratory in the Department of Anatomy and Cell Biology. My position as a laboratory assistant exposed me to the nature of scientific research. The experience was both technically and practically enlightening.

My project centered on determining gene regulation of microglia cells in a glaucoma model. Lab members gave me samples of RNA from different pairs of mouse eyes. In one eye, nothing had been done. In the other, an addition of a solution raised the eye’s pressure to imitate the effect of glaucoma. Half of these pairs came from normal mice, while the other half came from mice missing the P2X7 receptor. My goal was to determine whether the eyes imitating glaucoma showed any upregulation or downregulation of relevant genes, and whether there was a difference without the P2x7 receptor. Once RNA was converted to cDNA, data analysis of quantitative polymerase chain reactions (qPCR) was used to determine expression levels of inflammatory genes.

I was trained in a variety of technical skills by Wennan Lu (an experienced lab manager), Keith Campagno (a PhD candidate), and Huen-Yee Tso (an undergraduate student). I am now able to independently measure RNA concentration, convert from RNA to cDNA, run qPCR and analyze its data, and run agarose gel electrophoresis. I have also learned how to calibrate machines like an osmometer or a 7300 Real-Time PCR system, read programming relating to imaging, use a microscope to take photos of fixed cells, change cell media, and make various solutions. Prior to working in Dr. Mitchell’s Lab, I had no experience working in a lab. Now, I can see myself working in a laboratory beyond graduation.

In addition to new skillsets, I gained a lot of knowledge. First and foremost, to understand what I and other lab members worked with, I needed to do some research on different concepts in biology. Even after researching, the more experienced lab members still had to walk me through the more complicated concepts and experiments. Secondly, I learned what it is to be in a laboratory environment. Despite having our respective positions and individual projects, all of us who worked in the lab were a team. When I had questions or concerns, there were plenty of sources of guidance, especially the insightful advice from Dr. Mitchell. Even when facing discouraging results, I never once felt alone in my efforts. I realized quickly that laboratory work is not about clear-cut answers, but rather about developing and addressing meaningful questions. My position felt nothing like work. Every day was a new learning experience and a chance to contribute to furthering research. I am grateful to have been welcomed as a mentee and cannot express how impactful this experience was.