Author: Guillermo Tellez Major: Poultry Science

Hello, my name is Guillermo Tellez and I am a Poultry Science major. Over the course of the spring and fall semester of 2018, I have been working in Dr. Sami Dridi molecular biology lab to establish a correlation between the effects of heat stress and autophagy in the intestines of chickens. Autophagy is the process by which the body breaks down old or mis-folded proteins, fats, and other macromolecules into their building blocks to be reused. It’s a kind of recycling process that allows the body to be more efficient with the resources it has at its disposal. I have shown in a previous project that heat stress has a negative effect on the tight junctions holding the intestinal epithelial together and allows content from the lumen to spill over into the bloodstream which can cause a variety of problems including septic shock which can lead to death. In this project, I wanted to observe if chickens increase the amount of autophagy in their intestine to mitigate the effects of heat stress.  

In order to try to establish a correlation, I have used real time qPCR and western blot data to measure gene and protein expression. Working with both techniques has its challenges. Both techniques are very sensitive and require a lot of patience. During the RNA extraction phase for my PCR data, my RNA pellet became contaminated with genomic DNA (gDNA) and it took months to remove enough gDNA to get accurate results. Western blots are often joked to require a bit of voodoo magic and many scientists have their rituals when conducting them. A professor of mine always has a cup of coffee while staring at the machine from behind a window as it takes pictures of his blots for good luck. From the start they are hard. I struggled with smeared blots for a long time and everyone in my lab was baffled until an outside scientist enlightened me by telling me I had to remove the mucin from my proteins samples as the gut contains very high levels for protection from pathogens. Once this issue had been resolved and I had my blots, the next problem was with antibodies. Since chickens are not a popularly studied animal, it is hard to find antibodies that bind specifically to chicken antigens and so for the most part, we must hope that an antibody will cross react with the specific antigen of a protein we are interested in. After a couple months, in which I performed my own preparation ritual, I had collected all the pictures of my blots I needed.  

After I analyzed my data and having discussed them with my advisor and my lab technician at a weekly lab meeting, I have concluded that, under my defined parameters, there is no difference in the amount of  Autophagy-related protein (ATG) 3, ATG 7, ATG 16L1, Beclin-1, AMPK, mTOR, LC3A, or LC3B gene expression in heat stressed chickens when compared to chickens kept under thermal neutral, or normal temperature, conditions. Heat stress also has no effect on the protein expression of heat stressed chickens for ATG 7, ATG 5, Beclin-1, ATG 16L1, LAMP-2, phosphorylated mTOR, phosphorylated AMPK, AMPK, mTOR or ULK1 in the presence of heat stress. Though my hypothesis was disproven, I am happy with my work. Though there were days I was so frustrated with my project I wanted to give up, I pushed through. This being my second project, I have come to realize that in science “it is what it is,” but that I love research for the same reason I am training to run a marathon. I like the struggle. It forces me to push myself and grow as a person.  

I have decided to pursue a career in research. I believe that it will be a great outlet for my curiosity and love the ability to travel that comes with presenting your research. I also like being able to have discussions and arguments with brilliant minds about topics that I care about. In order to pursue this career, I am applying for the Rhodes and Gates Cambridge scholarships to conduct a master program in England where I could conduct research with some of the smartest people in the world. Until then, I have applied to the DAAD RISE to conduct another research project over the summer in Germany to improve the CRISPER/CAS9 gene editing system by taking epigenetics into consideration.