Monthly Archives: October 2017

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.

Stressed Rats and a Stressed Undergraduate

Chronic stress leads to a greater likelihood of the development of many conditions including post-traumatic stress disorder (PTSD), anxiety, irritable bowel syndrome (IBS), functional dyspepsia and other gastrointestinal (GI) dysfunction. This indicates a likely rearrangement of neural pathways and regulation, although the mechanisms of how this happens are not yet known. As an APS Undergraduate Summer Research Fellow, I worked for ten weeks under my research mentor Dr. R. Alberto Travagli studying the neurochemical oxytocin’s role in stress adaptation. My project focused on the regulation of oxytocin signals between the brain and GI tract under conditions of chronic stress in rats. In other words, I studied whether oxytocin has a different effect on the brain and gut of rats after they have been stressed.

Following a 5-day stress treatment on each rat, oxytocin was microinjected in the dorsal vagal complex (i.e. the brain area that directly signals the GI tract). The response to these injections on gastric tone and motility in two areas of the stomach were then recorded and analyzed. The research is still ongoing, but we hope to answer a few questions: How does the regulation of oxytocin change after stress adaptation? Does oxytocin work through different neural pathways after homotypic stress (i.e. same stress each day) or heterotypic stress (i.e. different stress each day)? Since females have a greater likelihood of developing GI disorders, do sex/estrogen levels affect the regulation of oxytocin under stress? Although we are still collecting data, I am very excited to see the results when completed and honored to participate in this research!

What surprised you most about working in the lab?

Upon starting this project, I was surprised by how much skill is required to complete the tasks at hand. Although the technology we use to inject oxytocin and record the gastric response is quite advanced, it can easily be faulted by a human mistake. For example, if I did not suture the sensors tight enough to the stomach, the responses were difficult to read and interpret. There was a huge learning curve to carrying out the research day-to-day and then it was another challenge to ensure I was as consistent as possible between animals. Additionally, I was surprised by how much my project changed between the beginning and end of the summer. For example, early on we injected a new pathway-blocker out of curiosity, expecting it to have little to no effect on oxytocin injections. Surprisingly, however, in one treatment group it seems to be blocking the effects of oxytocin. After, we used that pathway-blocker for every animal and its effects may be crucial to our final conclusions.

I am very grateful to the American Physiological Society for providing me this opportunity because it has made me realize how challenging a career as a basic research scientist is! This summer has exposed me to how exhausting, long, and physically demanding lab research can be. But, I love the big-picture parts of research; designing the experiments, analyzing the results, and adjusting when results are not as predicted. It is amazing to work on research that could be part of a bigger solution (i.e. understanding of anxiety/IBS/colonic pain), especially when you can collaborate with other researchers and pool data to come to even more conclusions within each study. However, I will admit that lab research is grueling work and, like the rats, I was a little stressed at times! I look forward to next year’s summer project so that I can experience translational or clinical research and gain a more holistic view of the research world.

Julia Zimmerman is a sophomore majoring in Neuroscience at Swarthmore College in Swarthmore, PA. She is a 2017 Undergraduate Summer Research Fellow (UGSRF) working in Dr. R. Alberto Travagli’s lab at Pennsylvania State University College of Medicine in Hershey, PA, funded by the APS. After graduation, Julia intends to pursue work as an MD-MPH, or MD-PhD, bridging basic research into the clinical world.

Understanding Muscle Force During Cyclic Movements: Does Titin Play a Role?

During cyclic every day movements, such as running, jumping, and walking, our muscles go through cycles of shortening and stretching. While there has been extensive research on muscle function for the last 50 years, there is no current muscle model that can accurately predict natural movements. For example, when active muscle is stretched, it produces more force than expected based on current theories of muscle contraction. Likewise, when active muscle shortens, it produces less force than predicted by current theories. For years, scientists have been measuring properties of muscles under highly controlled conditions. The classic force-velocity relationship shows that force generated by a muscle is inversely related to the velocity of the shortening. However, this relationship changes during natural, more life-like movements. Recent work suggests that for a given velocity, muscle force is higher during cyclic contractions than the traditional force-velocity relationship. My research investigates the role of the elastic protein titin in the force-velocity relationship measured under different conditions. Using a mouse model with a mutation in titin, I conducted in vitro muscle experiments to compare the force-velocity relationship in cyclic and controlled (isotonic) conditions. Hopefully, my results will shed light on titin’s role as a spring in active muscle. If titin truly does store energy like a spring, this could account for the extra force and lack of force in the stretch-shortening cycles. This research will allow us to better understand movement on a whole organism scale, which can prove quite useful in prosthetic design and bioengineering, for example.

Much like the active muscle, doing research in a lab goes through cycles, except instead of stretch-shortening cycles, it is periods of challenge and reward. Some days, you go into lab, collect great data, and leave feeling utterly fulfilled. However, other days, you go into lab and it seems as though you spent your entire day trouble-shooting. Mainly though, our experiments worked and we were able to collect useable data. We have yet to fully analyze our results, but preliminary results seem to support our expectations.

In general, I have found my lab group experience to be very similar to my experience with playing college soccer. Both activities involve a group of people working toward a common goal. While in soccer, your team is working together to win, in the lab, there are many scientists working together to uncover a truth. Collecting and analyzing data is a collaborative effort and, to me, that was the best part of summer research. Working as part of a lab team allows you the opportunity to constantly learn and build off of others. It teaches you to adapt, be open to new ideas, and to use your time efficiently. The worst part of day-to-day life in the lab, is that sometimes data collection does not go as planned and you need to figure out what went wrong. However, this aspect doesn’t seem so bad when you have your lab team to help brainstorm.

Overall, my time in the lab has been an incredible experience. It has helped me grow as both an individual and as a scientist and has stimulated my interest in future research opportunities. It is an experience I would highly recommend to other undergraduate students!

Lindsay Piwinski attends Pitzer College in Claremont, CA. She is a 2017 Undergraduate Summer Research Fellow (UGSRF) doing research with Drs. Jenna Monroy (Pitzer College) and Kiisa Nishikawa (Northern Arizona University, Flagstaff, AZ). She hopes to attend graduate school in the future and continue pursuing research.

Impact of Injury on Inflammation

For my research project, I will compare the levels of a known marker of inflammation in and around motor neurons in rats with and without cervical spinal cord injury. We will examine rats with chronic C2 incomplete spinal cord injuries, and compare them to uninjured tissues. We will be examining the frozen and preserved tissue under a microscope to quantify the different levels and locations of inflammatory markers at the different time points. The results of this experiment are important because they will enable us to better treat those who have suffered from the devastating effects of spinal cord injuries. This experiment is necessary to determine if p38 MAP kinase (a specific known marker of inflammation) is activated following spinal cord injury. The results will allow us to determine how to proceed in our search for successful rehabilitative treatments for patients. If p38 MAP kinase is activated following injury, it may call for future studies to investigate treatment of this specific cause of neuroinflammation in order to improve the outcomes of the rehabilitative treatments our lab is studying. Neuroinflammation can decrease the positive effects of rehabilitative efforts, and therefore is something we need to study so we can reduce this inflammation and better treat those who are suffering from spinal cord injury.

Life in the Lab

My experience in the lab was very educational. I did not realize how many different things actually go into research. I became familiar with behind-the-scenes tasks that you do not realize need to be done. For example, we spent a large majority of our time sectioning tissue, which I never realized would be such a large and time-consuming part of the experimental process. In addition, I became independent in many different techniques and procedures used in our lab. I learned proper animal handling and care, the methods used for immunohistochemistry, as well as proper imaging strategies and techniques on the Keyence microscope that we use in our lab. I also learned how to problem solve when problems arose. I found that one of the biggest challenges in research is how time-consuming and detail-oriented everything is. It is necessary to plan very far in advance and plan other aspects of your day around what is needed in lab. I was studying for the MCAT while completing my research project this summer, and I found it very difficult to dedicate time to studying. However, I quickly learned to manage my time wisely and study during down-time in lab and in the evenings. I believe this skill is not only valuable for research, but will also help me throughout my life.

Although what I did day-to-day varied, there were certain tasks that remained constant. For example, animal care and running exposures was something that needed to be completed by someone every day, so I was usually around to help out with those two things. In addition, the weeks were organized in a way that there usually was not more than one big task going on at a time. For example, there were weeks focused on surgeries, as well as others focused on perfusions and harvesting. What I did between rounds of animal care throughout the day varied depending on the week and what needed to be completed. Some days were filled with staining, while others dealt with microscopy. In my opinion, the best part of working in a lab was how often you were able to see your hard work pay off. Although the experiments tend to take at least a few months to complete, there are many milestones where you begin to see the outcomes of all of your hard work. Personally, I thought the hardest part of my time in research was not having set-in-stone days. Your schedule can vary every day depending on the point of the project you are in and what needs to be done, so you need to be able to adjust your plans and schedule around your lab responsibilities.

One of the best parts about working in a lab is being a part of a large team with a common goal. It is much more rewarding to accomplish a goal when everyone is working on it together in my opinion, and it’s nice to always have people around who are willing to help you and your project succeed. Research is usually not a one-person job, and for good reason; there is so much that goes on to ensure that a project is successful, and everyone in lab is needed. Throughout my research experience, I have developed a deep appreciation for how important research is to the functionality of many different aspects of society. Advancements in everything from technology to medicine would not be possible if scientists were not working hard in lab each and every day, and I am glad I will be able to take this appreciation for research into my future career. Research is a tough task, but it is truly life-changing in more ways than one.

Ashley Holland is an undergraduate at the University of Florida in Gainesville, FL. She is working in the lab of Drs. Gordon S. Mitchell and Elisa Gonzalez-Rothi at the University of Florida under the UGREF fellowship funded by APS. After graduation, Ashley hopes to attend medical school and use her skills acquired during her research experience to further the medical field and help her patients in new and innovative ways.

In the Lab for the First Time

Throughout the summer, I have been participating in what is my very first research experience. Apart from learning and doing techniques for the first time, the biggest challenge this summer has been trying to explain what I have been doing for the past month and a half to people that have asked me. My host’s research laboratory works on trying to understand the molecular mechanisms behind heart diseases, specifically working with the mitochondria. This organelle, also known as the “powerhouse of the cell”, is the one responsible for producing energy inside the cell. My project is focused more on mitochondrial dynamics, the fusion and fission processes the mitochondria go through to maintain their relative stability, and the proteins that facilitate them. I have been working to measure and quantify the presence of these proteins in different populations of mitochondria; and recently, I have been checking to see if these proteins become acetylated in samples that have gone through ischemia/reperfusion. Ischemia/reperfusion injury, also known as IR, occurs when the heart tissue is deprived of oxygen and nutrients; then goes through a period of reperfusion, when blood rushes back into the system. This sudden re-oxygenation period causes damage to the heart tissue. If fusion and fission proteins have become acetylated, inactivated due to the addition of an acetyl group, they could be affecting the stability of the mitochondrial population and possibly furthering damage during IR injury.

Life as a Scientist

Being that this is the first time I’ve worked in a research laboratory, it’s been great to finally have the chance to apply some of the knowledge I’ve acquired in my classes to something tangible; or understanding how various subjects I’ve learned from are applied in real life. I’ve spent the last six or seven weeks learning and improving my techniques in order to measure protein expression. In practices and even in actual runs, we’ve gone over procedures that take pretty much a whole day in order to eliminate possible errors with results. Currently, we’re working with the data we’ve gotten to see if we have to run more experiments in the next few weeks.

I guess the most surprising thing about this summer has been that the life as a scientist runs as a normal nine-to-five workday. Sure, there have been some instances that we stay later than usual or come in earlier to get things done; however, I had imagined something very different I suppose. Working as part of a lab team has been a great experience, mostly because you learn to have responsibility as a scientist and a coworker. Many of the areas and materials are used by everyone in the lab, so it becomes your responsibility to keep everything organized and stocked. You tend to think about leaving things in the order that you would like someone else to leave them. Apart from that, helping anyone out in the team goes a long way, because at the end of the day, we are all working towards the same goal.

Roberto Guzmán is a sophomore majoring in Cellular Molecular Biology at the University of Puerto Rico, Río Piedras Campus. This summer, he is working in Dr. Sabzali Javadov’s lab at the University of Puerto Rico, Medical Sciences Campus. He is a Short-Term Research Education Program to Increase Diversity in Health-Related Research (STRIDE) Fellow funded by the APS and a grant from the National Heart, Lund and Blood Institute (Grant #1 R25 HL115473-01). After receiving his bachelor’s degree, he plans to obtain a MD and continue into health related research.

Impact of Soybean Oil High Fat Diet on Hypothalamic Feeding Circuits in Mice

It is well known that consuming fatty foods induces obesity, although relatively little attention is given to the effects of different types of fat. The neuroendocrine hormone leptin is known to reduce body weight and fat through inhibition of food intake and increased energy expenditure (Friedman, 2011). Leptin’s actions are mediated by leptin receptor (OB-r), which is expressed in the arcuate nucleus of the whole hypothalamus (Meyers et al., 2009; Swanson and Sawchenko, 1980). Leptin may rapidly inhibit food intake by altering the secretion of hypothalamic neuropeptides such as neuropeptide Y (NPY), a stimulator of food intake, and/or corticotropin-releasing hormone (CRH), an inhibitor of food intake. (Jang, et al., 2000). STAT3, a neuronal transcription factor involved in the hypothalamic leptin signaling pathway via OB-r is necessary for the effects of leptin on food intake and hepatic glucose metabolism (Buettner et al., 2006). In this study, male C57BL/6N mice were fed one of four iso-caloric diets for a duration of 17-27 weeks: a high fat diet (40%) made up of coconut oil (HFD), a high fat diet containing soybean oil (SO) which is high in polyunsaturated omega-6 fatty acid linoleic acid (LA-HFD), and a vivarium chow control (VC). We hypothesize that transcription levels of OB-r and STAT3 will be down regulated in HFD mice, and that transcription levels of NPY and CRH will increase in the HFD mice. Pre-designed oligonucleotide primers for GOIs will be used and optimized for efficiency and specificity. RNA will be isolated from whole hypothalamus brain samples from each diet using an RNA isolation kit. Gene analysis (qPCR), will be performed to measure transcription levels of each gene. We have been able to cut and section whole hypothalamus brain samples from each diet, and have run efficiencies on the OB-r and STAT3 primer. Our efficiency results so far have shown OB-r and STAT3 primers giving good melt curves and efficiency values that lie within our acceptable range of 90-110%. The next steps will be to run efficiencies on NPY and CRH, and afterwards run qPCR with the isolated RNA samples. This work will give further insight on the regulation of specific genes in different types of diet.

Overcoming Challenges

Going into this project, I had prepared to dedicate a lot of my time to being in lab. I was responsible for my own schedule, which allowed me at times to enjoy some shorter days in lab, while understanding that some days would require me to stay longer to get the required amount of work done. Often, things would not go as planned, as some of the experiments did not yield expected results. When that occurred, I took a step back for a closer look at all the possible sources of error in the experiment. Often, these errors were shown in the experiment’s results, so the source of error was more easily tracked. Sometimes, however, these errors resulted in having to change different aspects of the project. For this current project, we had to change some of the primers that we were using because some of them did not yield good band/efficiency results. Luckily, while this did change what primer was initially going to be explored, it did not change the region or the overarching hypothesis of this project.

Going into lab every day has changed my work ethic greatly. Being able to set my own schedule for experiments has been extremely nice for me, and has allowed me to see personally the progress I am making in my project and the experience I am gaining in lab, which is one of the many pros of being a scientist. This experience has made me more aware of the importance of setting my own schedule, and how flexible it needs to be to account for possible errors in the experiment and setting up various meetings with professors, lab mates, and technicians, which can be considered a con of being a scientist. Experiments will often not give good results, and it may sometimes be frustrating to have to go back and troubleshoot each individual experiment. Having lab mates there, however, was a huge help in overcoming some of the challenges I faced with my experiment. Working as part of a lab team has shown me that each member of the lab collaborates with their peers’ projects and helps each other when necessary. I have learned a lot about each of my lab mates, including their project layout and work ethic. Working together with my lab mates this past summer has allowed to connect with them and gain experience from their guidance and help.

Edward Truong is a junior majoring in Cell, Molecular, Developmental Biology at the University of California in Riverside, CA. He is a Short-Term Research Education Program to Increase Diversity in Health-Related Research (STRIDE) Fellow working in Dr. Margarita Curras-Collazo’s lab at the University of California in Riverside, CA. Edward’s fellowship is funded by the APS and a grant from the National Heart, Lung and Blood Institute (Grant #1 R25 HL115473-01). After graduation, Edward plans to attend medical school and pursue a career as a physician.

The Effects of Anti-Depressants on the Developing Tadpole Brain

How do our bodies form the brain, its most complex organ? Developmental neuroscientists seek to answer this question by studying both the genetically-driven processes and the external, environmental forces that can intervene as well. Exploring how the environment can affect our brains through chemical, physical, and other stimuli opens new avenues for understanding, treating, and perhaps preventing several prominent neurological disorders. This past summer, I studied how exposure to the common antidepressant fluoxetine affects the brain during development. While children typically do not receive antidepressants, expecting mothers experiencing depression or having a history of depression are often prescribed such medication to resolve their symptoms and protect the fetus from the established harmful effects of stress. However, several case studies have shown that administration of fluoxetine is associated with the development of autism spectrum disorder (ASD) later on in the child’s life. Here, we seek to understand how fluoxetine exposure influences brain development at the neuron, circuit, and network levels. By using tadpoles, we can not only simulate a biological equivalent prenatal exposure, but also perform both behavioral and electrophysiology experiments to assess autism-like behaviors and individual neuron properties, network connectivity, and the population distribution of different types of neurons (excitatory, inhibitory, etc.).

My electrophysiology setup

Electrophysiology comprised most of my research and of my troubles! I performed whole-cell patch clamp recording, meaning that I recorded from the inside of a single neuron from the tadpole brain. This technique allowed me to study a wide variety of characteristics about the neuron, but was an extremely difficult method to learn. My first few weeks consisted primarily of learning the technique as well as troubleshooting my electrophysiology setup or “rig”. It took both creativity and persistence to return to the rig each day, ready to fix whatever problem occurs next and continually hope to acquire data. With the completion of this fellowship, I see research undoubtedly in a more realistic light. Rarely was it that experiments work perfectly. Good scientists are those who return to the lab bench with new ideas and an eagerness to learn and collaborate with others.

In fact, each day of my experience as a scientist this summer was marked by learning. Whether it was learning a new experimental technique or reading new journal publications or simply hearing about the more senior lab members’ tales in electrophysiology, I was always learning in the lab. Lab meetings were the highlight of the week, as we would all give updates on our individual projects that led to fascinating discussions for future directions. Above all else, this fellowship has imbedded the importance of collaborative learning within a diverse group of scientists. We all have our unique contributions to give, and I hope to continue expanding my own throughout my academic and scientific career.

Karine Liu attends Brown University in Providence, RI . She is a 2017 Undergraduate Research Excellence Fellow (UGREF) doing research in Dr. Carlos Aizeman’s lab at Brown University. In the future, she hopes to attend medical school and ultimately specialize in neurology or neuro-oncology.

Organohalogen Pollutants Causing Neurodevelopment Errors from Endocrine Disruption

My project is about an organohalogen, called polybrominated diphenyl ethers (PBDE), found in many flame retardants in the United States. I am studying this particular compound because it has been known to show disruption of the endocrine system in the body. PBDEs are known to be dispersed into the environment because over time they separate from the flame retardant mixture, usually present in dust particles, making it easier for inhalation. PBDEs mimic a hormone called vasopressin in the brain. Vasopressin pathways may regulate paternal behaviors, aggression, and associations in monogamous relationships between individuals. In previous studies, PBDEs were found in breast milk and are believed to increase externalizing behaviors such as hyperactivity, abnormal social memory, as well as repetitive behaviors associated with Autism Spectrum Disorder (Hoffman et. al). I am studying if there is a elicited phenotype within the generations of mice offspring who have been exposed to the chemical through gestation and lactation. I will be putting the offspring through behavioral tests to test their social behaviors, sensory-motor skills, as well as their anxiety/ depression. I expect the offspring that received the higher and lower dosage of PBDEs will show more hyperactivity and less anxiety, while the control group will show a normal phenotype found in most mice. This project is important because California has had the highest levels of this chemical in the United States. This chemical is not covalently bonded to its surfaces, causing it to be collected as dust and can be inhaled by humans. Studying the effects of PBDEs can bring more of an understanding to how there is neurodevelopment disruption and in turn help create an awareness of this toxicant and its effects on the body.

Life in the Lab

Being able to do research in a lab has been quite exciting. I enjoy running experiments and putting to practice the material I have learned in lecture. I learned many new techniques through the relationships I have built while working in this lab such as perfusions, neuroimaging, brain cutting and mounting, as well as how to run a Polymerase Chain Reaction (PCR). I was also able to teach my colleagues how to do some of the experiments I do for my project. I have been volunteering at this lab for almost a year now and am accustomed to the length of certain experiments and how much work goes into research. I have conducted many behavioral tests to examine the sensorimotor skills and anxiety of the mice. My behavioral tests have had a significant trend within different treatment groups. The results of my anxiety tests were skewed; this could have been due to some changes in lighting or previous stress on the mice. I expected to see that the higher and lower dosage of the PBDE offspring would have less anxiety in both males and females. However, this was only true for the males. The females’ data was more scattered and less predictable, while the control group showed more anxiety, which, is a normal characteristic in most mice. I have an upcoming experiment doing this anxiety test again and hopefully the data will be less skewed and follow the trend I am looking for in the PBDE dosed offspring.

My time spent here at the lab has actually been very enjoyable. I have been working six days a week (Monday-Friday and Sunday mornings). I come in at about eight in the morning every day to dose my animals and check their daily food and water intake. I then prepare to do my behavioral experiments and neuroimaging. When I am not doing these experiments my colleagues show me how to do some of the experiments they are working on. One of the best parts of working in this lab is the friendships I have gained and the connections I have established with the other lab I am collaborating with on my project. It has helped me to learn more about the different protocols and the differences between my lab and theirs. One of the worst things about being a scientist is when you come across a problem you are not sure how to solve. Asking others has helped so far, but not everyone understands my project and it becomes irritating. Despite that, being a scientist has been very enjoyable for me and it is making me consider going into research as a profession.

References

Hoffman, Kate, Margaret Adgent, Barbara Davis Goldman, Andreas Sjodin, and Julie L. Daniels. “Lactational Exposure to Polybrominated Diphenyl Ethers and Its Relation to Social and Emotional Development among Toddlers.” Environmental Health Perspectives 120.10:1438-1442, 2012.

Laura Anchondo is a Junior at the University of California, Riverside in Riverside, CA. She is a 2017 Integrative Organismal Systems Physiology (IOSP) Fellow working in Dr. Margarita Curras-Collazo’s lab at the University of California, Riverside. Laura’s fellowship is funded by the APS and a grant from the National Science Foundation Integrative Organismal Systems (IOS) (Grant #IOS-1238831). After graduation, her plans are to go into either pharmacy school or graduate school.

Neural Networks of Hypertension

Chronic high blood pressure, also known as hypertension, affects one in every three adults in the United States and nearly 1 billion people worldwide. It has been shown that the hormone, Angiotensin II (ANG-II), acts within the brain to stimulate the sympathetic nervous system, leading to hypertension. A region in the brain that is particularly sensitive to ANG-II and is involved in hypertension is the subfornical organ (SFO). The SFO is connected by neural projections to the paraventricular nucleus of the hypothalamus (PVN), and this pathway has been suggested to be important in cardiovascular regulation. The hypothesis of my study is that endoplasmic reticulum (ER) stress in the SFO-PVN projecting neurons is a cause of ANG-II hypertension. To test my hypothesis, the study will be split into two portions. In the first set of experiments, we will use staining of brain tissue (immunohistochemistry) to evaluate whether infusion of ANG-II causes ER stress in SFO neurons that send projections to the PVN. If we find ER stress in these neurons, we will use genetic approaches (CAV2-Cre-GFP and AAV2-Flex-GRP78) to inhibit ER stress and determine whether this also prevented ANG-II hypertension from developing.

What did you learn working in the lab?

Working in Dr. Young’s lab has been enlightening, exciting, and frustrating at times. There are many skills that take a great deal of practice to master that are required for studies like this one. For example, in order to perform Immunohistochemistry to identify markers of ER stress in the brains—a technique I learned this summer—I first had to learn to use a cryostat to precisely slice and collect the brains onto glass slides. I sliced dozens of practice brains before I mastered the skill and even now after weeks of practice, I still face challenges and make mistakes while performing this task. I also learned skills like suturing, small procedures, injections, bench work, and many others.

I very much enjoy the lab environment. It is great to have such a close-knit team that genuinely wants to help each other succeed. I wasn’t anticipating the level of team work involved in running a research lab, but that is truly the foundation. I would say that the friendships are the best part of working in a lab. The worst part of working in the lab is the waiting. For example, I surgically placed ANG-II osmotic pumps in the mice and then had to wait two weeks before taking the next step in my study. During that time, I practiced on the cryostat a lot, but there was still significant down time that I filled with tasks unrelated to my project such as helping other lab members with their work, cleaning, etc.

Melanie Judice is a senior majoring in Biology at the George Washington University in Washington, D.C.. She is an Undergraduate Summer Research Fellow (UGSRF) working in Dr. Colin Young’s lab at the George Washington University. Mary’s fellowship is funded by the American Physiological Society. After graduation, Melanie plans to continue research in the Young lab while pursuing entry into the medical field.