Working in Professor Snow’s lab has been eye opening for me in so many ways. It allowed me to truly understand what it means to work in research and how the root of successful work is to allow yourself to make mistakes, ask the difficult questions, and to be creative. This creative aspect of science really caught my attention. Before this experience in the lab, creativity is not one of the first words that would pop into my head when I thought about STEM or science in general. While science is revolved around fact and truth, discovery of science and the exploration of it is derived from a place of creativity. Scientists need to be able to ask creative questions and come up with new and inventive ways to test these questions.
This new understanding of science has inspired me immensely for my upcoming sophomore year. I am looking at my upcoming science classes with more eagerness, as I am genuinely excited to learn the material and to understand it completely so that I can be a creative scientist who can ask real questions. My role as a student is more than to just successfully complete my courses; I, as a student, am eager to understand this material, and I cannot wait to apply more of my scientific understanding in a practical lab setting. Working in the lab has also altered the way that I look at myself. I definitely feel that I can call myself a bona fide scientist. In the past, I always saw myself as a STEM or pre-med student, and never really felt that I could truly call myself a scientist. I feel now that I have earned that title, and I wear it proudly.
Throughout this research, I have learned in depth how to perform RNA extraction, reverse transcription, and qPCR. I have also learned about the Nosema ceranae infection that plagues honey bees, as well as the physiology and eusocial behavior of honey bees. Professor Snow also taught us about their specific cellular pathways in depth. Some of the pathways we learned about were the Unfolded Protein Response (UPR), Heat Shock Response (HSR), and Oxidative Stress Response (OxSR). This lab has definitely made me completely positive that I want to major in Cellular & Molecular Biology at Barnard, and I couldn’t be more excited about it. It was incredible to learn about these pathways because it gave me some perspective about just how complex and magical organisms are. These pathways are present in all eukaryotes, and it is just mind-boggling to me to understand how many components there are in a single pathway, and that there are countless amounts of incredibly complex pathways in all eukaryotes. It is also intriguing to me to think about how these pathways interact with each other in these systems. In the Snow lab, we are asking how these pathways, specifically Heat Shock Response and Oxidative Stress Response, are related to one another. We also wish to look into if these pathways differ with the age of the honey bee. For example, we intend to research younger nurse honey bees and older forager honey bees so that we can compare how these pathways affect the expression of certain small heat shock protein genes. My latest project has been to look into which genes the honey bee expresses when exposed to oxidative stress in order to see if the genes they express are the same or similar to that of Drosophila. We tried to induce the oxidative stress response by treating the honey bees with Paraquat as the oxidative stressor for either 6 or 24 hours. We then performed RNA extraction and qPCR to analyze which genes were expressed as a result of this.
Currently, I am working on performing qPCR on honey bee midgut samples that were treated with Dithiothreitol (DTT). DTT is a reducing agent and is supposed to cause an imbalance in redox components of the honey bee systems. Therefore we want to try and induce the oxidative stress response through the treatment of DTT to see if it causes expression of certain small heat shock proteins. By testing this, we will have a better understanding of the characteristics of the oxidative stress response in the honey bee, specifically in terms of what genes are expressed as a result of its induction.
The poster session this past Friday was such a unique and rewarding experience. It was a very prideful moment to be able to see our poster printed out and to be able to actually explain our project that we have spent so much time on to people. It was so much fun to describe our project to people who were equally as intrigued and invested in science as we were. Having the opportunity to do this made me realize just how different the roles of teacher and student are. In the past, it has generally been my job to learn the material that my teacher would explain to me, and it was interesting to have the tables turn and to be the one who was sharing knowledge with others who also have such a firm understanding of science in general. While I was able to teach what my project was, it still felt like a conversation with peers because we would bounce ideas off of one another and ask questions that would get me thinking about my own project in ways that I had never thought of before. It was also such an amazing experience to see what our peers have been working on this summer. I became even more proud to be part of STEM and this amazing scientific conversation with such intellectual women.
A main lesson I have learned from working in the lab is being able to learn from my experiences and to have the confidence to carry on even when I make a mistake. Mistakes should not be something to be ashamed about, but rather they should be embraced because I can honestly say that I have never made the same mistake twice during the time that I have worked in this lab because I have remembered what to do next time because of them. Mistakes are a major stepping stone to becoming a better scientist, and while they may be difficult in the moment, they always become a learning lesson later on.
I am so grateful to NOYCE for funding me to be part of this amazing lab. This summer has gone by so quickly, and I have learned so much about both science and about myself as a scientist and student. This experience has instilled within me an even more fervent enthusiasm for the upcoming semester and all of the science classes I will be taking. I sincerely hope to continue being part of the Summer Research Institute and NOYCE next summer.
These past few weeks, I’ve really felt that I have been able to narrow in on my project, and the accumulation of completed protocols, samples, and successful data has contributed to my current feeling of satisfaction and efficiency. In my project, I have become very comfortable and familiar with the RNA Extraction and qPCR protocols as I have completed both procedures numerous times for all of my samples. My samples consist of the head, gut, abdomen, and thorax of several nurse and forager bees which have undergone one of two different treatments. By using these samples, we are able to examine the expression of specific heat shock genes from four different tissues that we commonly look at. By specifically targeting nurses and foragers, we wish to determine which type of bee experiences a lower expression of these heat shock genes, and thus which type of bee has a lower overall heat shock response. Nurses and foragers are at different stages of their life cycles. Nurses are younger bees which care for the larvae, and foragers are older. By looking at the expression of these heat shock genes in both ages of bees, we wish to understand whether or not heat shock is more prevalent in older or younger bees. The primers used for the qPCR for this aspect of the project are actin, which acts as the control, and Hsc 70-4 (Hsp 68). The latter is a gene involved in the heat shock response. Working on this aspect of my project has made me very comfortable with the RNA Extraction and qPCR procedures, and my results have been very pleasing and encouraging. I used to be very tentative, and as the data improved every time I completed these protocols, I began to feel increasingly comfortable and confident in what I was doing. I feel excited to continue doing this to go through all of my samples and create a cohesive spreadsheet with all my data.
Another aspect of my project is creating double stranded RNA to perturb these heat shock response genes in the honey bee. I am learning how to set up gel electrophoresis, and how to isolate the bands in the dark room in order to eventually create double stranded RNA. Working in this lab, especially working with the double stranded RNA, has caused me to realize something about myself. Before SRI, I always thought of myself as a conscientious, organized, and careful person, however I am now realizing the degree of meticulousness, dexterity, and care that science requires. I can feel myself becoming a better and more efficient scientist everyday, as I am remembering to be even more conscious and focused on every little thing that I do in the lab. These qualities that I already thought I had are being improved every day.
We have been experimenting with newborn bees lately. The initial intention we had in using newborn bees was to potentially run longer term experiments with them since a newborn bee would be able to last longer than an older bee. It is also easier to handle newborn bees because they are too weak and young to fly or sting. We were able to easily manipulate them into cages by hand, and conduct our tests on them. Some problems arose in using newborns, and we needed to brainstorm and come up with innovations to our technique in using the newborns. One of these problems was that the newborns would eat little to none of the food that we provided them, because in that stage of life it is not really necessary for them to eat too much. This was a problem for us because in order to conduct our tests on the newborns, we use the food as the means to infect them with microsporidia or treat them with a certain drug, like Halofuginone or Dithiothreitol. We found a method that we could use to make sure that the newborns would eat. Because newborns can neither fly nor sting, we decided we could literally hand feed each and every one of them using a micropipette. While this did work and we were successful in feeding them with the treatments, it was incredibly time consuming. Another problem that arose was that a very large percentage of them would die very quickly. They wouldn’t live long enough for us to be able to observe the effects of our experiments. We thought that the reason for their high death rate could be because we were handling them too much, and that in their young and fragile state, picking them up by hand to both put them in their cage and try to feed them individually could have been too stressful for them. In order to try and resolve this problem, we decided against the hand feeding and to also try and simply pour them out of a box that held them into their test cages so we do as little manhandling with them as possible. We are currently seeing if making these tweaks to our methodology will be beneficial to achieve what we want. This process taught me how science is all about trial and error. It is common that something does not go as planned or work out in your favor, and when this happens you need to be able to ask the important questions and find creative ways to resolve any problems that may arise.
I have been doing a lot of spore counting with the samples we collect regularly from bees that we dissect. The purpose of spore counting is to record which bee midgut samples are infected and with how many spores. We do this using a hemacytometer. With the information gathered from spore counting, we make food for the bees we are testing on. Professor Snow taught us how to make dilutions with a positive tube containing the spores and sugar water to use to infect our bees. It has been really helpful to understand how to do this because it is central to many of the tests that we run in the lab. The same approach is used to incorporate the drugs and double stranded RNA to the food for the bees.
I have found the process of completely finishing an experiment all the way through to collecting and analyzing my data very rewarding and satisfying. Professor Snow has taught me how to input my data on a spreadsheet and to look at the numbers and try to analyze and figure out exactly what it could mean. I have learned that so much can be derived from numbers, such as which procedure along the way may have skewed certain data points in one way or another, or what the relationship between related data points may mean in the large scheme of things. Being part of this lab and having my own project really makes me feel that I am learning about all parts of science, including the actual lab work as well as the analysis and understanding of the results. My goal for the upcoming weeks is to continue with my project and to finish up all of my samples to analyze the data as a whole, as well as to get started on the poster describing our projects.
This summer I have been working in Professor Jonathan Snow’s lab, which concerns researching the N. ceranae microsporidia infection in the honey bee. So far, this internship has contributed to my understanding of STEM in more ways than simply teaching and reinforcing my biology knowledge and education. It’s given me the perspective of what it looks like to run a lab, and what the dynamics of the research world are. It has become so much more clear to me how many options there are in the STEM world, and it has been so exciting to explore it. Prior to this summer, I had the privilege of working in Professor Snow’s lab for the Spring semester of 2017. It has been so great to be able to apply the processes that I learned to conduct in the lab during the school year to fulfill an entire experiment. I also really appreciate how during the summer, I have the opportunity to be part of the experiments from beginning to end, starting from when the experiment was nothing more than a question or an idea. During the semester, I learned so much about lab work, specifically in cellular biology labs, but there’s nothing like being here during the summer to oversee and be partly responsible for entire experiments at a time. By having the privilege to do and experience this, my appreciation for STEM, more specifically biology, has increased dramatically. I am all the more confident and excited with my decision to major in Cellular & Molecular Biology at Barnard. It has also been such a gratifying experience to have been able to apply all the knowledge that I learned in my introductory Biology courses from my first year in a practical way. Not only has that knowledge been reinforced by recalling these principles of biology in the lab, but also by thinking about them in new and creative ways in order to help come up with new tests and experiments to answer our specific questions about the cellular pathways and immune systems of honey bees.
In Professor Snow’s lab, we have been exploring the effects of certain treatments on the unfolded protein response (UPR) in honeybees. Some of the chemicals we have administered to the bees, or have learned about, include Tunicamycin, Fumagillin, and Halofuginone. We also have been looking into how these certain drugs or the spore infection of N. ceranae, affect the expression of specific genes in the honeybee, such as the genes encoding the proteins actin and Hsp70. By looking into this, we have become increasingly familiar with the cellular pathways of honeybees and how they respond to these various treatments. We have been learning how to do several different processes in the lab. For example, to understand and explore different genes and their expression in both the honeybees and the spores, we learned how to conduct RNA extraction and quantitative polymerase chain reaction (qPCR). We also have learned how to set up microscopes and identify whether or not a honey bee’s midgut is infected with spores during spore counting. The results of our tests are always discussed as a group in order to share our own ideas of what these results may indicate, and what possible next-step tests and experiments we could perform in order to answer our questions in this research. I’ve definitely improved in analyzing and interpreting the graphs, charts, and tables of our results, as well as in seeing how and if our results are statistically significant, or if they happened by chance. We analyzed the data from one of the experiments which tested how the chemical Dithiothreitol (DTT) affected the spore count in infected honey bees.
From our charts, we were able to extrapolate that treatment of DTT for longer periods of time actually increased spore count within honey bees. Professor Snow has all of his honeybee colonies on the roof of Barnard Hall, and we make daily trips there in our bee suits in order to inspect the colonies and collect bees with aspirators for our upcoming experiments. We actually made our own individual aspirators out of plastic pipe and mesh, as well as the bee cages, which are used to hold the bees in the incubator and have a built in area for feeding. I have learned so much about the lifestyles and physiology of honeybees by doing this research and by being able to see the colonies in action. For example, during the honeybee colony inspections, we have been able to see the queen, the workers, the foragers, and the drones, as well as how honeybees store their honey, and what they look like in their stages as a larva and pupa. I also have learned how to dissect the honeybee midgut, as it is this portion of the bee that we look at to analyze the spore infections.
A unique experience in Professor Snow’s lab has been to read a collection of peer-reviewed scientific papers regarding tests related to the research that we are conducting. We read one of Professor Snow’s own papers, which talked about the UPR of the Xbp1 gene in honeybees. After we read this paper, we were able to discuss it as a group, focusing on what the paper was highlighting as well as what could be some possible next step experiments based off of its findings. Professor Snow also drew the entire Xbp1 cellular pathway of the honeybee on the chalkboard and explained to us in detail what exactly was happening physiologically. He described how exactly the honeybees respond when both DTT and Tunicamycin enter the system, and how they trigger the UPR. He went into detail about the phosphorylation proteins in the membrane of the endoplasmic reticulum and how, in this pathway, the Hsc70 proteins act as both chaperone proteins and as negative feedback regulators.
A goal that I have for the upcoming couple of weeks is to begin my individual project assigned to me by Professor Snow. My individual project concerns trying to find an alternate signaling system to the heat shock factors (HSF) of N. ceranae. Though N. ceranae are sensitive to heat, they do not regulate through HSF because they do not have them in their genome. Despite this, they do still have a way to up regulate proteostasis genes in response to heat. Some possible ways that N. ceranae could still up regulate gene expression without HSF could be through an alternate signaling system: for example, through genes skn7 or hms1 instead of HSF genes. To test this hypothesis, we could perturb the skn7 or hms1 genes with double-stranded RNA to see whether N. ceranae are still able to upregulate protein homeostasis after disruption of these genes. In order to try to induce a response, we could perform a “Dose Response” by putting N. ceranae under differing temperatures between around 34-45 degrees Celsius for varying amounts of time (1-3 hours).