Tag Archives: IOSP

Do Birds Have a Butt-Brain?

This summer, I worked with Dr. Monica Daley in the Structure and Motion Lab located at the Royal Veterinary College just outside of London. I was working on characterizing the function of a specialized spinal structure in birds and some dinosaurs known as the lumbosacral organ (LSO). This organ is hypothesized to function as a balance sensor by responding to changes in cerebrospinal fluid (CSF) flow, or by sensing strain in associated spinal ligaments. At its core, the spine is just a bony tube that surrounds the spinal cord, with CSF sloshing around between these two structures. When a bird experiences a disruption in balance, the LSO may sense this and restore equilibrium with a fast spinal reflex, instead of sending the signal up to the brain. This work has implications in physiology, motor control, and evolutionary biology. Firstly, this is a novel balance sensation mechanism that will give us a better idea of how bipedal land birds can quickly overcome balance perturbations. Their motor control strategy may be applied in robotics to create more efficient and resilient locomotor systems. Additionally, the LSO is thought to be important in the transition from dinosaurs to birds. Better characterizing its structure and function may shed light on this key evolutionary event.

 

The everyday life of a scientist can vary widely across labs and research projects. One of the many reasons I love biomechanics as a field is the sheer scope of techniques that I get to employ on a daily basis. While the core of my project involves simulating CSF flow on a computer, often times I find myself breaking up my day with hands-on work as well. For instance, I might begin a simulation in the morning, but conduct a balance perturbation experiment on a live guineafowl while the program runs. The next day I could be dissecting and staining specimens, commuting into downtown London to run CT scans, 3D printing an experimental apparatus, or writing code for numerical models. I get to feel like a biologist, mathematician, and engineer all in the same day.

 

Research is not always as glamorous as it is portrayed in high school and college courses. I often found myself frustrated with code or software that wasn’t working as I thought it should be, for seemingly no reason. Additionally, the results that you generate are often very difficult to interpret at a first glance. A large amount of my time was spent troubleshooting, writing data analysis scripts, and consulting colleagues to help develop intuition for the physical phenomena I was observing in my fluid dynamics simulations. Despite these struggles, I always felt inspired and motivated by the fact that I was conducting research that had never been done before. The reason why conducting sound experiments and interpreting data is so challenging is because it is literally uncharted territory. The intellectual rewards of making progress in a research project are very satisfying and often outweigh the struggles of achieving them.

 

Ethan Wold is a rising junior at Brown University in Providence, RI where he is majoring in Biology and Geophysics. At school, he works with Dr. Thomas Roberts studying the biomechanics of muscle and connective tissue in a variety of organisms including frogs and turkeys. This summer, he worked in Dr. Monica Daley’s lab in the United Kingdom studying the biomechanics of balance sensation in bipedal land birds as an IOSP fellow. After graduating, Ethan plans to do some traveling and conservation work, and then pursue a PhD in biomechanics or bio-inspired engineering.
Lovebirds—How Dopamine Helps Zebra Finches Fall in Love and Stay in Love

The concept of love has fascinated Homo sapiens since we first started experiencing those warm mushy feelings. Ever since then, painters and writers and actors have found avenues to express the ways that love makes them feel. Now, scientists are discovering what happens in the brain that creates the compulsion for monogamy that many believe Hallmark cards tricked us into thinking we needed. For my project, I am studying the changes in the brains of zebra finches that lead to monogamy. Of course, what we know as “love” has been socially constructed by humans, so technically I am studying what scientists call “pair-bonding” in monogamous birds. Research in voles, another monogamous species, has shown that there are increases of dopamine receptors in the part of the brain associated with rewards when voles have been pair-bonded for long amounts of time. Dopamine receptors bind dopamine—a neurotransmitter responsible for many feelings and movements, including pleasure—and send messages to other parts of the brain—for example, “This candy is delicious, let’s keep eating it!” or “I am in love with this other bird, let’s see if she is interested in mating!” Specifically, the number of dopamine 1 receptors increased the longer that voles were paired, and activation of these receptors also caused males to be disinterested in females that weren’t their mate—and dopamine 2 receptors increased when the pair-bonding was first happening (Aragona et al., 2006). Basically, the activation of dopamine 2 receptors caused the voles to “fall in love,” and the dopamine 1 receptor activation maintained the love over time. This connects to zebra finches because my PI, Dr. Coleman, has shown that by using chemicals that activate the dopamine 2 receptors manually, a scientist can induce partner-preference in zebra finches—sort of like a “love potion,” that makes zebra finches “fall in love”! The same method was also performed in Aragona et al.’s 2006 paper, where partner preference was induced in voles. This research is one of the final pieces in Dr. Coleman’s project about the mating preferences of zebra finches. You might be wondering—who cares about the love lives of zebra finches? I can name one famous person you might recognize who would be very interested in this research: Charles Darwin. This research furthers Darwin’s theories of sexual selection—what traits are sexually attractive, and should be passed down? How does the brain “fall in love”—in other words, how does the brain select a mate, one of the most important decisions in any organism’s life, especially if the organism is monogamous? Our preliminary results do not indicate an increase in neither dopamine 1 nor dopamine 2 receptors in paired birds compared to sexually naïve birds. Instead, we are seeing an increase in the enzyme tyrosine hydroxylase, which helps make dopamine. This result suggests that, in zebra finches, there may not be a change in the number of receptors, but instead just an increase of the neurotransmitter itself. We plan on continuing the experiment, and looking in several parts of the brain to see if there is an increase in a different area.

“The important thing to remember is that you have a goal, and that small hiccups shouldn’t dissuade you from research and making discoveries.”

Like Darwin, I will never just wake up one day and have all the answers to all the questions of the universe, much less innate knowledge of all techniques that I will need. For example, this summer, my lab has been collaborating with the lab of Dr. Stephanie White, specifically with her postdoc, Dr. Nancy Day at UCLA. We have been utilizing Dr. Day’s expertise in quantitative real-time PCR (qPCR). This is a technique used by scientists for many reasons, but in our case we will use qPCR to analyze dopamine 1 and dopamine 2 receptor gene expression in finch brains. This is one of the realities of research I learned this summer: rarely will there always be in your lab (or department) the materials or people who can do everything that you need to do, and sometimes you need to rely on your friends and colleagues (hopefully made by networking) and their expertise and equipment. To do qPCR, you need primers that can locate the gene of the receptor you are interested in in the genome. This is a very daunting task for a little primer! Recently, we have run into difficulties with our primers for dopamine 1 and dopamine 2 receptors. We have ordered new primers, and our next task is to determine which primers are the most effective at targeting this gene. We have also run into some problems with our cryostat, which is used to slice tissue. Fortunately, our department has a state-of-the-art cryostat, and we have learned how to use it, and it works spectacularly! Clearly, this is another reality of research: you win some, you lose some. The important thing is to remember is that you have a goal, and that small hiccups shouldn’t dissuade you from research and making discoveries. To quote the tagline of one of the best science fiction shows of the 1990s, “The truth is out there!”

I think the most important thing for people to realize about the day-to-day lives of scientists is that we aren’t discovering gravity or the cure to polio every day. Some days are a wash, literally—regularly deep cleaning our bird room is one of the great joys of research. But some days are great, like when you finally manage to perfect that technique that you have been struggling with for weeks. I feel like an expert with the cryostat and subbed slides—there’s no better feeling than doing something easily after practicing for months. Slicing tissue is like riding a bike—you never forget. It’s important to learn as much as you can from people who know more than you, but it’s equally important to teach. I think a lot of people tend to think that scientists isolate themselves in their lab, barely ever emerging for food or sleep, laughing maniacally whenever their experiment works. Actually, that sounds pretty accurate, now that I have it all written out. But I guess scientists tend to not isolate themselves completely—we usually have a lab partner or a collaborator working with us. So it’s more like, you barely emerge from your lab for eating or sleeping, you laugh maniacally, and your lab partner is the last person you spoke face-to-face with in the past 3 days. Joy! (Of course, I’m being sarcastic. It’s been 5 days!). I have a new appreciation for the power of collaboration. And I think that is why the image of a scientist toiling alone in a lab bothers me so much—it ignores the fact that all scientists rely on others, and have others who rely on them. Perhaps if Frankenstein got his research peer-reviewed, his life would have gone very differently. As Newton said, “If I have seen further, it is by standing on the shoulders of giants.” Having stood on the shoulders of many giants myself, I look forward to the day that I can be a “giant” for another young scientist.

References

  1. Aragona, B. J., Liu, Y., Yu, Y. J., Curtis, J. T., Detwiler, J. M., Insel, T. R., & Wang, Z. (2006). Nucleus accumbens dopamine differentially mediates the formation and maintenance of monogamous pair bonds. Nature Neuroscience, 9(1), 133-139. DOI: 10.1038/nn1613.
Lauren Eisenman is a senior majoring in cellular/molecular neuroscience at Scripps College in Claremont, California. She is a 2017 Integrative Organismal Systems Physiology (IOSP) fellow working in Dr. Melissa Coleman’s lab at the W.M. Keck Science Department in Claremont, California. This summer, Lauren is funded by the APS and a grant from the National Science Foundation Integrative Organismal Systems (IOS) (Grant #IOS-1238831). In the future, Lauren plans to attend graduate school in a neuroscience program and pursue a career in academia.
My Summer of Science

In the summer of 2017 I spent my time working an unconventional full-time job in the lab growing as a scientist. I was given an opportunity to be mentored by an established research professor who I look up to greatly. She helped me step by step go through the process of coming up with a project, understanding the current research in the field, and guiding me through the behind the scenes documentation to get fully approved by local, state, and federal agencies.

My project that I am undertaking is geared towards ultimately understanding the roll of dietary choices and its implications on oxidative stress, leading to potential for cardiovascular disease. My lab and I are utilizing birds as a model to demonstrate the effects of a high-carbohydrate diet and its ability to cause a hyperglycemic condition. For the study, Mourning Doves will be used due to their high density in the Tempe, AZ area and much of the studies leading up to this used the same birds. They have a unique ability to combat the oxidative stress that is commonly seen due to high blood sugar. We hope to better understand how their body combats the negative effects of hyperglycemia in efforts to process the information regarding the protective mechanisms of the vascular system.

Realities of Research

The idea of moving research forward can be very attractive. The actual process of progressing it may not be as attractive. Research is a very meticulous, detail-oriented task that needs to be well planned, documented, and thought-out for it to properly work. There is no book you can read that gives you the exact answer to what you are doing. This is the world of the unknown and if properly done could potentially help shift the understanding for those to come after you. However, a major hiccup that I believe is frequently over looked as a young scientist going into a research lab is that it’s quite common to get negative results, or even though the data seems to support what you thought, it is not statistically valid based on the mathematical analysis. This is a huge part of research and can be very testing of resiliency as the process of getting this far could take months or years.

Life of a Scientist

Although, it is common to see science hitting the news headlines frequently on your phone and social media feed, it is a much longer process of meticulous work than what is publicized. In the efforts of using an animal in the aide of progressing the scientific community there is a very stringent process of approvals needed to work with them. For example, Mourning Doves are migratory birds, so my project requires an approval by the Arizona State Game and Fish Department, the U.S. Fish and Wildlife services, and the Institutional Animal Care and Use Committee (IACUC) before I can begin the research itself. I was surprised as I began to work in the lab by all the intricacies of administrative background work that went into being able to conduct research. While repeating experiments take a lot of time and effort, I now feel as if there is equal if not more effort done in paperwork and formalities than physical experimentation. I, myself am still waiting to begin the physical hands on portion of the research. I have worked through the course of the summer gaining valuable experience in the creation of the protocol and understanding the approval process. I have also been given the opportunity this summer to work very closely with another research project that is much larger than my own.

This summer has been an invaluable experience in research that I would not have gained otherwise without the gracious efforts of the APS.

References

  1. Braun, and Sweazea. “Glucose Regulation in Birds.” Comparative Biochemistry and Physiology, Part B1 (2008): 1-9. Web.
  2. Smith, Toomey, Walker, Braun, Wolf, Mcgraw, and Sweazea. “Naturally High Plasma Glucose Levels in Mourning Doves ( Zenaida Macroura) Do Not Lead to High Levels of Reactive Oxygen Species in the Vasculature.” Zoology3 (2011): 171-76. Web.
  3. Sweazea, Braun, and Sparr. “Novel Role of Insulin in the Regulation of Glucose Excretion by Mourning Doves (Zenaida Macroura).” Zoology122 (2017): 58-62. Web.
Will Clark is a senior biochemistry student at Arizona State University in Tempe, AZ. He is a 2017 Integrative Organismal Systems Physiology (IOSP) fellow conducting research in the lab of Dr. Karen Sweazea at Arizona State University. His long-term goal is to become a Physicians Assistant, and join the health care workforce to help improve the health and well-being of others.
Post Spinal Cord Injury Female Mice have a Higher Concentration of Glial Cells

In the laboratory this summer, I studied one of the myriads of cell responses that occur in the spinal cord post-injury. The cells that I studied were microglia, which play a role in cleaning up debris and acting as an immune response. It has been found that post-SCI (spinal cord injury) female mice have better functional recovery than their male counterparts. Scientists studying SCI’s are currently researching possible causes for this difference in healing. My project focused on whether male or female mice have a higher microglia response post injury. The tissue I analyzed was collected from male and female mice 42 days post-moderate SCI. I first had to stain the spinal cord sections with eriochrome cyanine, which shows the degree of degeneration the spinal cord underwent post injury. Immunohistochemistry was then performed on the tissue, which is a form of staining that binds fluorescent antibodies to the cells you are looking for. Because of the fluorescence that is attached to the cell, you can then use light to illuminate the microglia and then image the cells. To get the most accurate comparison, I selected a portion of the spinal cord that I could identify in every animal and quantified the cells in just that area. My data concluded that female mice have significantly (p: .0187) less microglia.

Realities of Research

This was my first time taking part in research in a professional lab setting and there were a few things that surprised me, but for the most part I knew relatively what I was getting into. I think the most surprising thing was that there was a lot of sitting around and waiting for either results or for my slides to dry. As a student that works better when pressurized, I feel that this aspect will be the biggest barrier keeping me from going into a research career. There were also a lot of issues that I had with the tissue itself and aspects that hindered my ability to analyze my results.

My responsibilities changed from day to day, whether I was sitting at a desk and reading for hours straight or doing a stain. I also spent a lot of time learning about lab equipment and how to safely complete an experiment. I think the best part was when I was able to work by myself and take ownership of my own project. However, there were many times that I was lost or confused and really needed help. The worst part of this research was all of the down time that I had. Working as a lab team was very interesting because there were always people around that had something interesting to contribute to my project. However, there were also issues with finding the specific person that could help you with a part of your project.

Julie Wilson is a senior majoring in Biology and Chemistry at Baldwin Wallace University in Berea, OH. She is a 2017 Integrative Organismal Systems Physiology (IOSP) Fellow working with Dr. John C. Gensel at the University of Kentucky in Lexington, KY. Julie’s fellowship is funded by the APS and a grant from the National Science Foundation Integrative Organismal Systems (IOS) (Grant #IOS-1238831). After leaving Baldwin Wallace University, Julie plans to attend medical school and pursue a career in pediatrics working with queer and diverse youth.
Turkeys and Treadmills: Identifying Gait Transitions in Grounded Running

The basics of locomotion must be understood to fully comprehend not only the actual movement that takes place, but the mechanical aspects behind those exact same movements to gain more knowledge as to why an animal uses a certain method of locomotion.  Animals move with different gaits, whether it’s flying, swimming, running, and probably the most commonly used method of locomotion in humans – walking.  For this reason, my project deals with observing turkeys while they run on a treadmill to characterize the transition that takes place from a walk to a run by measuring stride length and stride frequency. A stride is the action that occurs when a single foot moves from start to finish and usually takes place in two steps for bipedal or two legged animals. Stride frequency is the number of steps in a second by a single foot, and stride length is the maximum distance that occurs in a single stride. While running, humans experience an aerial phase, which is when both feet are off the ground at the same time. Turkeys are unique in this aspect in that they instead partake in grounded running, or running without an aerial phase.  Turkeys may not seem like the most ideal animal model, but have been proven to be useful for studies that are interested in running mechanics and energetics to define principles that apply to plenty of other animals.

Realities of Research

Whenever an experiment is performed, every single aspect is not going to proceed the way it’s supposed to – no matter how much meticulous planning is involved. There were days when the turkeys were completely cooperative, and other days where obtaining footage wasn’t at all possible. Getting turkeys acquainted with using a treadmill for the first time is not their favorite activity, to say the least, but they have grown accustomed to it to some extent. I’m still in the process of obtaining data, but I hope to gain a clear understanding of the mechanism turkeys rely on when they shift gears from a walk to a run. What ultimately has surprised me so far is the amount of software and equipment available which makes, not only this project possible, but also other experiments that are being conducted by other lab members, and provides valuable information to understand the mechanics of turkey muscle. Despite running into usual situations when conducting an experiment, what I didn’t anticipate of this project was eventually learning how difficult it would be to handle turkeys to begin with.

Day-to-Day Life of a Scientist

The day to day life of a scientist definitely has its benefits and setbacks when going through an experiment and actually being a part of a lab. Being a part of a lab team is intriguing in that over time you gain a better understanding and appreciation for what these people have been and currently are working on, and provides for a great learning opportunity whichever way you look. The down side of life in the lab is working on a project and motivating yourself to analyze and interpret your data as to what, and more importantly to formulate an explanation for the phenomena you have observed. Of course, all is not lost since you are given the chance to discuss what you’ve found with your lab members, but ultimately it comes down to what you think is going on. Although this may be true, pursuing research is not just a one-time occurrence but rather a giant learning process. Over time, you learn or figure out how something works, but that same answer leads to even more questions that you didn’t think of before. You may not always find the answer to begin with, and that in itself may feel defeating. Every obstacle is and should be seen as a learning opportunity, and despite all the unanswered “why” questions left in the world, research is there to help us create a roadmap to finding answers to these questions, no matter how small an answer may seem.

Karina Vega is a rising Senior majoring in Biology at California State University, San Bernardino. She was one of the undergraduates selected for the 2017 Integrative Organismal Systems Physiology (IOSP) fellowship, conducting research in the lab of Dr. Thomas Roberts at Brown University in Providence, RI. Karina’s fellowship was funded by the APS as well as the National Science Foundation Integrative Organismal Systems (IOS) (Grant #IOS-1238831). After graduation, Karina plans on obtaining her Ph.D. in comparative physiology in hopes of pursuing a career in academia.
Learning from Obstacles in Science

The Western diet is high in fats and sugar and can lead to an increase in metabolic diseases, which cause a chronic state of peripheral inflammation (1). My project this summer aims to observe the effect of diet on brain inflammation. We used a mouse model of tagged peripheral monocytes (3). Monocytes turn into macrophages, which target inflammation in the body and brain (2). These mice were fed either a diet high in fat and fructose or a normal diet for 5 weeks. Then the blood from their brain was washed out, and the brain was sliced. The slices were stained for the genetic tag for the peripheral macrophages. Peripheral macrophages found in the brain suggest that chronic inflammation weakens the blood-brain barrier, allowing peripheral macrophages to cross where they increase brain inflammation. This may cause damage and may have links to diseases such as Alzheimer’s Disease and Parkinson’s Disease, which show increased inflammation in the brain (4). This project would further support the idea that a healthy diet could be a key factor in prevention of brain diseases.

This project, as well as science in general, had many obstacles that I had to overcome. Originally, I planned to analyze a different genetic mouse that modeled Alzheimer’s disease. Those mice were also going to be fed a high fat high fructose or control diet, and were going to be compared to see if there was an increase in peripheral macrophages in the brain in diet treated mice. However, those brains didn’t have the blood cleared from the brain, which limited our ability to see the stain. To overcome that problem, we used the new mouse type that had peripheral monocytes tagged, which had the blood removed from it. With this new mouse model, I would have a smaller number of animals, but I could better test my hypothesis.

I enjoy the day-to-day life in research. I was expecting it to be somewhat repetitive, but that was far from the case. I had many problems that I had to solve and was constantly learning, which made the time fly by. My day was broken up by working on different parts of my experiment, writing and reading literature, and meeting and talking with my lab members. The best part for me was that I constantly learned new things. There were many hiccups in my summer experience, which were disheartening at times. However, solving these problems and further learning more made it rewarding as well. The Tansey lab has many members who have been very helpful in solving these problems. I enjoyed being a part of a larger team, as there were so many projects going on that I could learn from.

References

  1. De Sousa Rodrigues, M. E., Bekhbat, M., Houser, M., Chang, J., Walker, D., Jones, D. P., Oller do Nascimento,C., Barnum, C. J. & Tansey, M. Chronic psychological stress and high-fat high-fructose diet disrupt metabolic and inflammatory gene networks in the brain, liver, and gut and promote behavioral deficits in mice. Brain, Behavior, and Immunity 59: 158-172, 2017.
  2. Khoury, J. E., Toft, M., Hickman, S.E., Means, T. K., Terada, K., Geula, G., & Luster, A. D. Ccr2 deficiency impairs microglial accumulation and accelerates progression of Alzheimer-like disease. Nature Medicine 13: 432-438, 2007.
  3. Saederup, N., Cardona, A. E., Croft, K., Mizutani, M., Cotleur, A. C., Tsou, C.-L., Ransohoff, R. M., & Charo, I. F. Selective chemokine receptor usage by central nervous system myeloid cells in CCR2-red fluorescent protein knock-in mice. PLoS ONE 5: e13693, 2010.
  4. Selkoe, D. J. The therapeutics of Alzheimer’s disease: where we stand and where we are heading. Annals of Neurology 74: 328-336, 2013.
Lindsey Sniffen is a senior majoring in Neuroscience and Behavioral Biology at Emory University in Atlanta, GA. She is a 2017 Integrative Organismal Systems Physiology (IOSP) Fellow in Dr. Malu Tansey in the Department of Physiology at Emory University in Atlanta, GA. Her fellowship is funded by the APS and a grant from the National Science Foundation Integrative Organismal Systems (IOS) (Grant #IOS-1238831). After graduation, she plans to pursue a Ph.D. in pharmacology, and then work in the pharmaceutical industry.
Atomic Force Microscopy on Titin, the Mega Protein

This summer in Flagstaff, Arizona at Northern Arizona University (NAU) I worked with Dr. Samrat Dutta in the Nishikawa Lab for biomechanics performing atomic force microscopy (AFM) on the N2A region of titin molecules at different pulling speeds in both the presence and absence of calcium. Titin is found in vertebrate muscles and is the largest known protein molecule (Nishikawa et. al, 2011). Currently, we know that titin functions in passive muscle movement. However, it may provide an important addition to our current understanding of both active and passive muscle function (Nishikawa et. al, 2011). Understanding titin isn’t just revolutionary for muscle theory, Nishikawa’s lab is applying this new information to improve prosthetics. AFM is a non-conventional type of microscope (shown below) that allows us to record the stiffness and stability of biomolecules such as titin by pulling on its spring-like domains. The titin is chemically attached to a surface and the AFM traverses that surface and records changes in its topography using a laser. This experiment allows us to predict the behavior of titin and its contribution to muscle force under different conditions.

Working with AFM has been a steep learning curve for me. AFM wasn’t a process that I was at all familiar with before this summer. With guidance from Dr. Dutta over the course of these 10 weeks I’ve learned about the chemistry, function, and potential of AFM. Unfortunately, we received low usable data yields and this may have been a result of the protein unfolding before the experiment began. As a result, there was a lot of tweaking of our methods to gather a larger set of more accurate data. The analysis of our data afforded me an opportunity to learn physics and chemistry beyond the scope of my university classes. However, we have not yet completed the analysis of our data. I look forward to seeing the outcome of our experiment and contribute to the ever-growing data on titin. Hopefully, my research will answer how much force titin can contribute and in comparison to previous works, does the N2A region of titin react differently than other regions.

What was it like working in the lab?

Dr. Kiisa Nishikawa’s lab group is filled with scientists doing various projects in all different disciplines of the muscle physiology field. Throughout my time at NAU, I had the opportunity to network and learn from all different kinds of people such as postdocs, graduate students, and full professors. They all guided me through my new environment at NAU and supplied me with both professional and scientific knowledge from their different disciplines. Working in a collaborative group meant having support during disappointing moments and always having someone to run ideas by. When the experiment wasn’t producing the amount of data we expected, a team of graduate students and postdocs helped my mentor and I brainstorm possible causes and solutions. This brainstorming session was how we determined that dialysis may be a useful alternative to our previous protein purification method. Working with a large team also means that you must share resources and space which, can make things more difficult. Overall, this summer was an invaluable experience in many aspects and it wouldn’t have been possible without the American Physiological Society (APS). I want to thank APS for allowing me and so many other undergraduates the opportunity to contribute to different fields of research.

References

  1. Nishikawa, K. C., et al. Is Titin a ‘Winding Filament’? A New Twist on Muscle Contraction. Proceedings of the Royal Society B: Biological Sciences 279(1730), 981–90, 2011.
Blair Thompson is studying biology at Scripps College in Claremont, California. She is a 2017 fellowship recipient of Integrative Organismal Systems Physiology (IOSP) funded by the APS and a grant from the National Science Foundation Integrative Organismal Systems (IOS). She worked with Dr. Samrat Dutta and Dr. Jenna Monroy at Northern Arizona University this summer. After graduation, she plans to attend medical school and become a physician.
My Summer Researching and Learning About Worms and Alternative Medicine

My research project explores the possible future uses of the alternative medicine Astragalus, which is a Chinese herb known for its immune stimulating effects, such as decreasing melanoma tumor growth. I am specifically looking for immune system responses in transparent roundworms called C. elegans. I am treating the worms with different concentrations of the Astragalus medicine, then observing how their immune system pathways have reacted. My research expands the information available on alternative medicines and their validity. This project could lead to further, more specific studies as well as studies on other animals and potentially humans. Astragalus could be proven to be a viable medication for disease or replace current cancer treatments and medications.

Working in a research lab has proved to be very interesting and exciting. I started out the summer unsure of myself and the techniques involved with my project. There were many standard procedures and instruments I needed to familiarize myself with to successfully and effectively perform tests. Over time I became more comfortable and could do most tests and procedures on my own, only after being instructed by my research host, of course. My research was unsuccessful at first as I couldn’t get the desired result from polymerase chain reaction (PCR) with the house-keeping gene actin. We reran the end-point PCR and ran another gel but still had no results. After taking another step back and rerunning the cDNA synthesis with my RNA sample, we finally had a successful PCR result. I know there are many techniques I have yet to master in my research journey.

I was surprised by how long certain experiments took and how much everyday maintenance was required to continue working with the worms. Much of my time was spent counting the worms for survival assays or synchronizing the worms to prepare for a test. Also, there was constant upkeep of Nematode Growth Media plates and E. coli stock. I also had to adjust to working independently on my research as opposed to working in a class. I had to prepare all the materials I needed and go through the steps on my own instead of having everything laid out for me. The best part of research was getting successful results and making progress, the worst part was becoming frustrated when results were undesirable or no results were produced at all.

Alyssa Knudson is a junior at Coe College majoring in Biology and Molecular Biology and minoring in Music and Chemistry. She is a part of the Coe College Cross Country and Track and Field teams. She is also involved in Pre-Health Club, Biology Club, and Intergenerational Connections at Coe and a member of the co-ed service fraternity Alpha Phi Omega. She has participated in a 10 week research program working under Dr. Cassy Cozine the summer after her sophomore year. After college she plans to go on to graduate school and get her degree in Genetic Counseling.
STZ Effects on HA Production and HAS Expression in Channel Catfish Through Liver Damage

My research project is primarily focused on how different doses of a chemotherapy drug, Streptozotocin (STZ), affect channel catfish. This drug has been commonly used to treat pancreatic tumors and induce hyperglycemia, or high blood sugar, in rodents. We are using channel catfish as alternative model organisms for investigating human metabolic disorders such as obesity and diabetes because these fish have an accelerated growth rate similar to the phenotype observed in obese and diabetic patients. However, administering STZ into channel catfish has been shown to display the opposite effect, resulting in a high mortality rate and hypoglycemia, or low blood sugar. Abnormal morphology of the liver and gall bladder has also been noted in past studies, possibly indicating liver damage associated with the STZ treatments. However, the exact mechanism(s) associated with the development of hypoglycemia and liver damage after the administration of higher doses of STZ has never been examined in channel catfish. My biggest contribution to this experiment has been to study the expression of the Hyaluronan synthases (HAS2 and HAS3), which are membrane-bound enzymes in mammals, in various catfish cDNA samples before and after injection of STZ. These enzymes are directly related to the production of Hyaluronan (HA) in the blood in response to severe tissue damage.

Channel catfish kept in aquariums inside the FHSU grounds department building. Photo credit: Megan Dougherty, Fort Hays State University

Realities of Research

Conducting research in a laboratory setting is very much how I expected. I had minimal pipetting experience from a biochemistry course I had previously taken, so I knew perfecting my technique should be first priority. This basic skill is so important in ensuring that the data I am collecting are going to be accurate and useful in the final analysis. I found it interesting that the assays we were conducting were extremely sensitive to the surrounding environment and realized the importance of keeping a clean laboratory space. A peer in my lab experienced some setbacks involving contamination in the blank tube of her PCR results. We were able to detect the contamination through a picture taken after gel electrophoresis, and we were then able to try to identify where the contamination could possibly be coming from. We eventually had to take a full day to clean the entire lab bench with ethanol and a bleach/water solution to try to get rid of the problem. There haven’t been any issues since the lab group took that step. My results in this experience so far have been as expected. I haven’t found an expression of HAS2 or HAS3 in any tissue samples collected from catfish before STZ treatment by just using the basic PCR and gel electrophoresis technique. This is not surprising. My mentor has explained that these enzymes would not be expressed in high amounts until after the fish have been injected with STZ because they are related to the repairing of tissue damage. Because low concentrations of HAS should be expressed in some healthy tissues, however, my mentor has just recently taught me how to conduct real-time PCR to visualize the results in a different way. Once STZ treatment is complete, I do expect to be able to visualize HAS2 and HAS3 mRNA expressed at a higher concentration, primarily in the liver.

Life of a Scientist

Overall, my research experience has been a very positive one. I have realized how important working as a team in a laboratory setting is. It is very useful to understand what your peers are experimenting on and be available to help them along the way. This leads to building strong relationships and a plentiful collection of data. I have also learned that it is important to stay flexible throughout experimentation and to understand that getting no results is not necessarily a bad thing. It just means that new options and techniques need to be explored to find exactly what you are looking for. My mentor has been great at keeping me and my peer motivated throughout this experience by answering any questions we have and helping us, whether the issue is not getting results or getting results that are contaminated. The best part about working in a lab is leaving each day feeling accomplished and knowing that I am learning so many new things. Another great aspect of my summer research is that I can feel myself becoming more confident with the procedures I have learned as time goes on. I find myself asking for less help and getting things done correctly in a timelier manner. The worst part about research is going through a lot of small steps and spending time on assays that do not show any results at the end of a long day. I sometimes feel as if time is wasted when this happens, but it is important to remind myself that no results still reveal something about the overall experiment.

Preparing to run gel electrophoresis. Photo credit: Abigail Schmidtberger, Dr. Kobayashi’s research lab, Fort Hays State University

References

  1. Nevarez E, Ordonez-Castillo N, Spainhour R, Kobayashi Y. Treatment with Streptozotocin (STZ) causes hypoglycemia and alters the stability of reference genes for real-time PCR analysis in the liver of channel catfish [Online]. The Official Journal of the Federation of American Societies for Experimental Biology. http://www.fasebj.org/content/31/1_Supplement/1014.3 [12 July 2017].
  2. Itano N, Sawai T, Yoshida M, Lenas P, Yamada Y, Imagawa M, Shinomura T, Hamaguchi M, Yoshida Y, Ohnuki Y, Miyauchi S, Spicer AP, McDonald JA, Kimata K. Three Isoforms of Mammalian Hyaluronan Synthases Have Distinct Enzymatic Properties [Online]. Journal of Biological Chemistry. http://www.jbc.org/content/274/35/25085.full [12 July 2017].
Megan Dougherty is an upcoming senior majoring in Biology with an emphasis in Health Professions at Fort Hays State University in Hays, Kansas. She is a 2017 Integrative Organismal System Physiology (IOSP) Fellow working in Dr. Yashiro Kobayashi’s lab at Fort Hays State University in Hays, Kansas. Megan’s fellowship is awarded by the APS and a grant from the National Science Foundation Integrative Organismal Systems (IOS) (Grant #IOS-1238831). After graduation, Megan plans to attend a graduate program in hopes of pursuing a career as a physician assistant or medical technologist.