writing from this seminar:

I was really nervous about applying to SURF, I didn't know what my chances of getting into the program were, but I know I needed the money. As I started getting rejection emails I was disappointed but began thinking about my plan B- I already contacted my PI and knew I was interested in his work, could I work for him for free and take on a part-time job? I was nervous because I knew this would be stressful and I would be exhausted every day. Luckily, SURF-MedEx reached out to me right before I was about to apply for a second job. I was in!

 

I reached out to my PI, Dr. Herman, and began working at the lab in early May- almost a month before SURF actually began. I had little lab experience and wanted to get accustomed to how this lab functioned and what kinds of lab techniques they used daily. Everything was working out in my favor. My only problem was that Dr. Herman's lab was on the UC Reading Campus- a 20-minute drive from my apartment. I did not own a car and was not in the financial position to buy one, so I had to take the bus. 

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The bus was a whole experience on its own. The 20-minute drive turned into an hour commute back and forth. The bus system was fantastic, but the people I met on the bus were not. At an estimate, I think I was catcalled at least twice a day. I learned to ignore it and keep walking, knowing that getting angry could be dangerous in situations like these. The bus drove through Avondale, a historically black and underprivileged area. Getting on the bus, I was always afraid and I couldn't help but wonder if my fear was a result of implicit bias or if my fear was warranted as a woman traveling alone. This experience in itself helped me reflect on my actions and my thoughts. 

 

The graduate student I worked under, Marissa, was incredibly sweet and began teaching me lab techniques almost immediately. I slowly began learning what the lab focused on in the brain, and how Marissa's research fits into the lab. Her research was about loss and the effect loss has on neurobiology. I found this extremely interesting and was eager to begin my own exploration.

 

It was very overwhelming- I heard a lot of jargon and it took almost a month for me to understand bits and pieces of what the lab meetings were about. There was another undergraduate student there, Ria, who was in my shoes last summer and would always check on me to see how I was doing. I am so grateful that she was also working at the lab and was able to almost "interpret" what everyone was talking about. 

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Marissa is very organized and is a great planner. Every week we would talk about our goals for the week and what we hoped to accomplish in the weeks to come. The first lab technique she taught me was cutting rat brains. I learned to use a machine called the Microtome which enabled us to cut very thin slices of the brain from front to back in order to examine the exact section of the brain we were focusing on. I began helping Marissa cut brains that were put through the loss model- enrichment removal. These brains were used in a pilot study, which is when the experiment is run on a few brains to test whether or not anything needs to be changed in the protocol.

 

The experiment that I learned was Immunohistochemistry, which is used to stain certain cells in order to see them under a microscope through fluorescence. As I was learning how to run an immuno, I slowly began to understand what I was doing. Marissa's work focused on the Basolateral Amygdala, and how loss could cause an increase in Perineuronal nets (extracellular structures) that could inhibit the brain from learning new information and creating new neural connections. The brains that we were using were put through an enrichment removal model (give rats toys, take them away). We were using a specific stain to stain CAMKII, which stains for activated cells in the Basolateral section of the Amygdala (BLA). The goal of this experiment was to see if more inhibition from the nets would cause less activation in neural cells. 

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Marissa had to attend a stress conference, so for two weeks, I came to the lab to practice my lab skills. Mounting brain sections onto slides was the hardest in my opinion, so I practiced and practiced, mounting at least three brains (two hours each) every day for two weeks. 

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When Marissa came back, I began a new project to work on parallelly as Marissa ran tests on the pilot study of CAMKII. This new study was looking at how the Single Prolonged Stress model, an animal model of PTSD, can affect the BLA. This study used a total of 20 brains, 12 of which were precut and eight of which I needed to cut. I began by running the immuno on the 12 already cut brains, then cut the last eight and ran the Immuno on them as well.

 

The hypothesis was the same as before- the stress model would cause an increase in nets and decrease the brain's ability to learn new information. Based on previous studies, we knew that the majority of Perineuronal nets surrounded certain cells called Parvalbumin Interneurons (PV). We also knew that PV was inhibitory in the BLA and decreased the fear signal that the amygdala sends out.

 

After running T-tests, we learned that our results showed the opposite of our hypothesis. As stress increases, the number of nets decreases. We also learned that the SPS model (the animal model of PTSD) showed a decrease in the number of PV itself. Since PV is inhibitory in the BLA, meaning it decreases the fear response that the amygdala sends out, a lower amount of PV would allow the BLA to send out a greater fear response. This ties in well with the human characteristics of PTSD, a hyperactive BLA which produces a hyperactive fear response. Through my project, we were able to explore and create preliminary data about the processes of the BLA during PTSD.

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This experience taught me valuable lab skills, such as using a Microtome to cut brain sections and running immunohistochemistry to illuminate certain cells. I also learned to stay organized and set weekly goals for myself. I learned that to reach my goals I need to stay dedicated and persevere, even if it means an hour commute on the bus to and from the lab. I learned to make the best out of my situation, finding books to read on the way to and from the lab, and finishing five books in the span of three months. Asking questions and being patient was key to gradually begin to understand some of the jargon that was being spoken at the lab. I also learned that this kind of work takes an incredible amount of patience.  The daily work is mundane and monotonous, and it could take weeks or months to actually see results from your experiments. 

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