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).