Amongst all of the hard work, we managed to find some time to spend on a vacation! Recently, many of the Howard Hughes Fellows (unfortunately, some had to go into lab during the weekend and couldn't come :( ) took a day trip to Wilmington, NC to enjoy the lovely weather and gorgeous beaches. Here are some pictures of the awesome trip:
My research question for this summer is:
“Are synaptic defects the first defects in neurodegenerative diseases?”
Before I started, the PIK3C3 gene was removed from mutant mice to model neurodegenerative diseases. The PIK3C3 gene is involved in membrane trafficking (exocytosis and endocytosis) and is necessary to maintain a functional neuron, so removing it from the mutant mice has understandably caused some neurological problems. We’ve already seen older mutant mice (9 months old) develop neurodegenerative diseases, with notable differences in morphology between the mutant and control mice of the same age. As the mice age, neurons begin to die in the mutants, and we would like to figure out why this happens.
The first step was to determine at what age the brains of mutant mice began showing morphological differences from the brains of control mice. I’m doing this through retrograde detection, so I’ll start with older mice and keep analyzing younger mice until I find the age at which there aren’t morphological differences between the control and mutant mice. I started with 3 month old mice. I sectioned the brains of two different mice, one control and one mutant. Then I stained them to analyze the abundance and organization of neurons and glial cells. Some symptoms that affect the mutant mice are neuron apoptosis and gliosis, so in a mutant mouse we would expect to see a greater number of glial cells and fewer or deformed neurons, when compared to the control mouse. This is what I observed in the 3-month-old mice, which means that even though they aren’t showing outward symptoms of neurological degeneration, their brains have already been affected.
Now, I’m going to take a look at the brains of mice that are about 1 month old to detect morphological differences. With mice this young, we don’t expect to see overwhelming differences between the control and mutant mice, so this is the age during which I will be looking for synaptic differences. We’re hoping that the only difference between mutant and control mice are synaptic defects in the mutants.
So why is this important? I’m going to use Alzheimer’s Disease as an example, because it’s common, but this applies to neurodegenerative diseases in general. When someone develops Alzheimer’s Disease, they show symptoms gradually. But even before the smallest symptoms are shown, their brain has been affected. By the time the symptoms are great enough to warrant a trip to the doctor and a diagnosis, it’s already way too late. The only thing doctors can hope to do is slow the progressions of the disease because it’s very, very difficult to reverse neurodegenerative diseases. If we can figure out what happens first during neurodegenerative diseases (again, we think synaptic defects come first), then we can begin the process of figuring out how to reverse/prevent/treat/etc. the disease.
Coming into this program, I had no idea what to expect. I had never done research in a lab before, so I really didn’t have a clue what it would be like. Even so, I had certain impressions of what lab would entail. However, it’s way different than what I expected. Firstly, it’s a very relaxed atmosphere. The researchers can create their own working hours, it isn’t the typical 9 to 5 job. They are in the lab for as long as they need to be to finish what they are working on. Also, I had the impression that research would be very fast-paced, racing to find the answer to a research question. While it is in a way, a lot of procedures take days to complete. I’m currently doing an in situ hybridization, and I won’t finish until Thursday or Friday. Some of the steps involve waiting for hours until continuing, or even incubating overnight. Because of this, I’m generally working on several things at once. Even so, there are some slow moments in lab, but there are also moments when I’m juggling several things at once.
Now that I’ve seen how research works (at least in the limited amount of time that I’ve been here), my expectations have formed in a different direction. Impressions created by labs in school lead a student to believe that if you follow the procedure you’ll have everything you need to figure out your question. Real research, however, is very different. It’s much more of a guessing game, trying different things out until something works and you can take the information you learned from that to ask new questions. For example, we’re running probes now to detect the layers in the cortex of both mutant and control mice at different ages. There isn’t a formula to determine what we’re looking for, no calculations to do in advance to save us time. It’s just a matter of patience, waiting until we find our answer.
This summer I hope to gain experience working in a lab and an accurate perception of what lab life is like. My goals go beyond learning how the equipment works or what protocols to follow for different situations. I’d like to learn how to ask the questions that lead to great research, to be able to interpret results and then take them to the next level. It isn’t as simple as following directions with guaranteed success, but more of a blind searching in hopes of finding what we’re looking for (or what we aren’t, that can be just as interesting). In other words, I’d like to experience what it means to be a scientist.
Hi! My name is Katie Budolfson and I’m a rising Sophomore at Duke . This summer I’m doing research in Dr. Fan Wang’s lab and my mentor is Dr. Liangli Wang. I’ve never done lab research before, so this is an entirely new experience and we had to start at the basics. It was all really confusing at first, because I didn’t know how to use any of the equipment, but I’m already starting to get the hang of things.
So I don’t know all of the details yet, but here are the basics. We’re modeling neurodegenerative diseases and analyzing the way membrane trafficking is affected by the PIK3C3 gene in mice. There are control (normal) mice and the mutant mice. I’ll section their brains, slicing them into very thin layers, and place them on slides. Then I’ll stain the slides to show different things in the brain. For example (see picture below), this week I stained them with one stain to mark for nuclei (in blue) and another stain to mark for neurons (in green). We can then compare the control and the mutant mice brain sections to see how the neurons and their layering are affected by PIK3C3.
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And this is where I work:
