Mice Research with Dr. Minicozzi
I was quite nervous to begin research at MSU-Mankato and initially reached out to a leader in the Undergraduate Research Center office to get more information on how to start. They gave me resources on how to write an email to professors which gave me confidence in the next steps. I compiled my research of three professors who were working on projects that interested me. One of which I knew was retiring soon, so I kept her as a back-up. Another professor I had heard was not easy to work with and I was left with Dr. Minicozzi. I was worried he would not have space or deny me due to my lack of an individual and unique idea for a project. After sending the email, he responded quickly with a “Yes I have space, let’s meet soon!”. This surprised me and overwhelmed me since I assumed I would not start the research until the following semester. Although, I was excited and felt huge relief that I did not need to have to think about back-up professors. In the meeting he stated he would like to continue his research into the effects of perchlorate, a common pollutant found in explosives such as fireworks, in mice bodily processes after finding evidence of effect in his previous fish research. This seemed relatively interesting to me, I was mostly thankful I was working with mammals aside from fish or worms as I had heard about his other student’s study.
The first project I worked on with him studied the bodily processes including the gonads, thyroid, and muscles and how those are affected by perchlorate. I began setting up cages, each containing one mouse. The top portion of the cage had wheels zip tied, and I needed to connect the counters that recorded the wheel spins. This task proved to be time-consuming and frustrating. The wheel counters were not of the best quality and disconnected or were inaccurate. After numerous weeks and around ten hours spent cutting and reconnecting wires. I got around eight to work properly, there were 20 mice in the study of which the others were observed later. Now I could start to set up the waters and begin the 50-day cycle of the study. To set up the waters, I added 200 milliliters of water to each bottle. In the perchlorate treatment mice, I added 100 parts per million of perchlorate whereas for the control treatment I added zero parts per million of the chemical. Every five days within the cycle I recorded the number of wheels spins the subject made and their respected water consumption. Throughout the cycle, there were a few times when the wheels were disconnected due to the cages getting changed. This was frustrating because the data was then insignificant, and I needed to spend extra time that day to be sure the wires were reconnected and working properly.
At the end of the 50-day cycle I was trained to euthanize the mice and learned to dissect the portions we were going to study. We euthanized the mice with a plastic bin and gas, I released the gas slowly, so the animals are not in as much pain. The body portions I dissected included the gonads, thyroid, and legs. The ovaries of the female mice were particularly difficult to find because they were much smaller than the male testes. Once I had found a few ovaries and confirmed I was taking the correct part with my professor I became confident in my ability. Dissections were my least favorite part of the research. It smells and the morals of it all makes my stomach turn. I have learned to have more of a “farmers mentality” with the mice so I do not get heart broken when euthanizing but it was still difficult with the first few mice. To make things easier on myself I stepped out of the room while the gas was running, I think if I had heard any noises, I would not have been able to go through with the rest. Learning to dissect and euthanize animals in a human way is likely something I will need to do for any job I have in the zoology career field. Even in research jobs there may be an animal that is struggling or found dead, and a dissection is done to determine what happened to the animal. Additionally, having these types of fine motor skills could be beneficial in banding wildlife or inserting trackers for research into ranges or diets of the animals. All of that to say, I was thankful when I was finished with dissections, and I started a new project that was behavioral based.
After winter break, I asked Dr. Minicozzi what the next steps for the research would be and he said to start setting up new wheel counters for the other mice since we had only eight in the first trial period. I was about to start the adventure of connecting more wires and hoping they would work again. Dr. Minicozzi ordered more counters for me in hopes that they would help. I was excited when those arrived, and I figured I could set up the waters that week as well. Unfortunately, that was not the case, and I needed to cut wires and reconnect a bunch of them. In this 50-day cycle there was lots of incorrect data as the wheels and the wires got disconnected on a weekly basis. In this instance, none of the data was usable. This was difficult to process, especially since I had spent so much time dedicated to making sure the data was eligible for future use. I was angry that we could not use it but also relieved I did not have to go through the dissection process again. All the issues with inaccurate wheel counters have taught me that it may have been important to get a license of accuracy for the data collection. There are many caveats that can come with such certifications such as highly expensive products. Dr. Minicozzi told me there were better options, but they were each well over $1,000 for one counter. In the future when I am conducting my own research, if money allows, I will pursue the more expensive option. If I am limited to money from a grant for research or conducting such through a company, I would be sure to read reviews on the product to ensure accuracy and to not waste valuable time.
In the meeting where Dr. Minicozzi suggested the next steps to set up more wheel cages he also said he had another project for me to do alongside it if I was interested. In that moment, I said yes and interpreted his offer as he enjoyed working with me. At this point between research, work, and college I was already busy and his suggestion for a new project overwhelmed me once the thought of it settled in. This project involved studying the behavior and stress levels of the mice to compare perchlorate treated to the control. I used the mice that were already treated for the 50-day cycle and instead of euthanizing, I used them for the new project. I used a device called an elevated plus maze that Dr. Minicozzi had provided for me. This device is elevated about one foot off the ground and has two arms that are enclosed with walls as well as two arms that are wide open.
The first big challenge with the plus maze project was finding a room that was suitable for recording data. I recorded videos of each of the mice and they needed to be in a room without external stimulation that may affect their stress levels. When we found a room that was not in use there was a bunch of old office supplies that we needed to move out each time we recorded. This was very time-consuming and frustrating. This data will help to be a foundation for future, more representative data of this study. The beginnings of this project are being laid so in the fall we have the kinks worked out and can run data more smoothly. After finding a room and moving furniture to have space for the maze I placed a kitty pool in the middle of the room with the maze inside. Therefore, if the mouse jumped out, they would not be able to run the halls of the science building. The camera was placed above with a ring stand and some clips. Currently, there is a room in the Animal Care Facility I am using where I can leave the maze set-up in that room and save time in the set-up. This dedicated space also allows for consistency of each mouse. When I input the videos to the software program to analyze, the frame should be set up and not need readjusting for each field I have drawn on the maze. A camera is still placed above but with a tripod attached to the ceiling now. For both set-ups, I set up a Zoom meeting with my computer in the room and my phone connected. When I stepped out of the room, I could then watch the mouse to be sure data was recorded correctly. Recording data went smoothly other than the few times when a mouse jumped off the maze or decided to urinate in the maze. I cleaned the maze in between each trial regardless of mice defecation. This was to ensure the mice did not just follow the scent of the previous one. Data recording was just much more time consuming for this project due to the 30-minute acclimation period for the mice to get used to the room and the 10 minutes they spent exploring the maze. Although time consuming, I was much more interested in watching how the mice responded and behaved in this project.
The data analyzing process proved to also be time-consuming and frustrating. I have used my patience skills and learned how to be even more patient in the process. The computer program I am using allows me to set parameters and insert the videos I recorded to collect data such as time spent in each arm. This should allow easier and more accurate data collection instead of relying on the human eye. Neither my professor nor I know how to use the program though so there has been a lot of trial and error and asking other students for help. After countless hours of playing with the program we found the parameters we wanted, and the computer is collecting data correctly. Learning new things on a large scale like this will be something valuable for my future career. I have spoken to a few people that work for the Minnesota Department of Natural Resources. They have mentioned other data analyzing programs I am not familiar with that if I go on to do research, I may have to learn these computer programs on the spot. I have improved my patience skills through my research with Dr. Minicozzi which will help in learning new things in the future. My perseverance and drive to complete a project will also be helpful for possible future publications.
After analyzing data and researching how others set parameters for similar projects, I presented my findings with a poster presentation in the Undergraduate Research Symposium in April of 2024 as well as at the Undergraduate Biological Sciences Research Symposium, a separate and more specialized presentation for science faculty and I presented my conclusion for my results as I expected; the perchlorate treated mice tended to spend more time in the closed arms of the maze. Although, I did not have enough samples for the data to be significant and conclude official results. I am currently working to increase my sample sizes (Fall 2024) to ideally obtain significant results. These findings could be concerning and indicate a behavioral change that is potentially stress induced, but it will be good to raise awareness that the chemical has potential to affect humans. Therefore, more research could be done to protect us due to the ease of getting into our water ways.
I did not feel like I had all the knowledge that I needed going into presenting and believed you should be an expert in your field or project. When I got questions that I did not know the answer to such as “Does perchlorate penetrate through the blood brain barrier?”, I was nervous. I did not want to make up an answer with an expert asking the question. I answered this in a sort of “I do not know” way while also explaining that I did know previous research in fish has shown perchlorates effect on the thyroid gland. This gland releases a lot of hormones that could change behavior. I confirmed this with my professor a few days after presenting and he said a similar answer which gave me confidence. This lack of knowledge was the entire point of presenting, the goal is to learn more from colleagues and experts. They challenge your findings and how you could potentially improve.
Other lessons I learned when presenting for three hours included losing your voice and forgetting talking points. Water is important for these kinds of events but even with that aid, my throat was scratchy since I do not normally speak for that long. I also got nervous presenting for a few people, and it was hard to know who wanted more of a summary and who wanted to hear all the details. I missed details for some who wanted them, this is something I want to learn to work around. It might be simple to ask the listener what kind of presentation they want.
There was a difference presenting at the Biological Sciences Symposium as opposed to the Undergraduate Research Symposium (URS). At the URS I needed to explain more science in simple terms whereas at the Biological Science Symposium judges were graduate students in the department that were conducting similar research. In some ways it was easier to relate to the judges of the Biological Sciences Symposium, since they seemed to have a greater knowledge that expanded beyond my own. These experiences will directly shape my future career as well as the next few months. I will present at a National convention in January of 2025 with the Society of Integrative and Comparative Biology (SICB) in Atlanta, Georgia. I plan to address the questions I received at each of my presentations in January for SICBE. I want to include more information about the thyroid in my oral portion to better understand and explain how perchlorate effects the mice. In my future, hopefully a research-oriented career, I will be mindful of my presentation skills that I gained from these undergraduate experiences and extent of knowledge that may be important for larger presentations.
To sum up, I have learned and enhanced multiple skills in my professional personality through my research with Dr. Minicozzi. These include improving my patience with the process, developing more problem-solving skills using trial and error, working hard, and managing my time. I discovered how credited, high-quality materials can make data more accurate. This discovery also led to inquiry about how expensive excellent materials can be, a major barrier in research. In my first project, my data would have been more credible with more expensive materials but there was not enough money in the budget. This research and presentation have given me the foundational knowledge of the first steps to data collection and processing to creating visuals and presenting. This will allow me to be more efficient and proactive with projects in the future.
