I am very happy to be back on the 6th floor of Altschul to continue conducting research this summer. I have been working in Professor Sever’s lab for the last year - starting as a student in SRI in 2016 and continuing through the academic year for credit. I am working on a project with the goals quantifying RNA and protein levels of molecules related to Alzheimer’s disease.
This project has been passed down through many students and I am very excited to continue and progress the work. Specifically, I am investigating the mechanism connecting miRNA expression and excess metal ion treatment with hyperphosphorylation of tau through the MAPK pathway and the cleavage of APP in AD. In the Sever lab, we carry out cell culture and treat out cells with small molecules or metals, as with my project. To see the effects of the treatments, we extract the RNA or protein of interest and quantify it. On the first day, I immediately took cells that we had previously stored in a freezer and “brought them up from frozen”. To do so, I needed to thaw the frozen cells, plate them in media, and then store in an incubator until I am able to work with them. After the cells start to grow to a preferable density, or confluency, I split and plate them in new petri dishes. Since a few passages are necessary to begin working with the cells, I am waiting for the cells to grow and reading journals in the meantime.
The project I am working on requires me to recall lessons about gene expression from my classes: the Spring semester of Intro Biology as well as Mendelian and Modern Genetics. Gene expression is the process of using information from a gene to synthesize a functional gene product. There are many steps involved in gene expression. The three main ones are DNA replication, transcription, and translation. Transcription results in RNA, especially messenger RNA (mRNA), and translation results in protein. When planning experiments, specifically the timing of experiments, I need to take into consideration these processes. For instance, translation is towards the end of the long process of gene expression so I need to wait 24 hours after treating cells with metals until I can extract protein from the cells. Likewise, I needed to find the optimal time for extracting RNA from the cell. I conducted a time trial to find the time which resulted in the most mRNA available in the cell since mRNA can degrade overtime if not used.
In addition to learning the more tangible side of research, I’ve also learned the more emotional side of research. For example, I’ve learned new techniques and procedures in lab and from journals, but I’ve also learned that a lot of time experiments don’t work or don’t produce a result you were expecting. There is a lot of problem solving involved in lab work once you get poor or unexpected results. In July of last summer and during the last academic year I was troubleshooting a procedure. I was trying to quantify protein using the Western Blot protocol. After the very long procedure which typically takes a few days, I was not seeing any protein bands when imaging. We went back to the drawing board and tested all the steps in the method which could possibly cause the result we were getting. Finally, we were able to better optimize the procedure and see protein. What I hope to do this summer is finish quantifying the proteins of interest.
Along with quantifying protein, I hope to work a step prior in the gene expression process and quantify RNA. I hope to see a similar response with specific mRNA to that of the protein they encode. For instance, if there is an increase in MAPK3 protein after treatment compared to that of non-treated cells then we would hope to see the same increase for mRNA for MAPK3. I hope I will be able to quantify the molecules of interests and be able to replicate the results at least three times. I will also be working with three different neuronal cell lines and I hope that I can replicate the results in all three.
I also plan to quantify the amount of microRNA in the cells after treatment with excess metals. MicroRNA are involved in the gene expression process and specifically can cause translational repression. The miRNA complementarily binds to mRNA and inhibits translation so no protein is produced. There are many miRNA that are related to Alzheimer’s disease. I have been reading a lot of journals to find more information about miRNA and have compared all my sources. There is a lot of information available to us and it is important to check for any new findings or inconsistencies.
Each week, my group presents our research updates in a group meeting. I have learned that presenting is just like telling a story. It is easy to jumble up the specific details you want to communicate, but it is necessary to take some time to plan and present the information logically giving plenty of background information and details. Having presented in group meeting for about a year I have gained confidence in my ability to present and communicate the work I have done. Looking back, I have noticed my presentation skills improve from just my experience over last summer. I was floored by the eloquence of the presenters when I listened to many thesis presentations in the Chemistry Department. I am glad that I can seem calm as well as experienced when presenting.
I have also learned that it is okay to ask questions. I try to reason concepts out by myself but often need to check for confirmation with Professor Sever. Often, Professor Sever answers my question by stepping me through the process in question and drawing out information from me that I can add to solve my problem. I have found this method of answering questions may take a bit of time, but it is very helpful to have the person asking the question learn the answer by working it out with some guidance. This was one of the techniques that Barnard College suggests peer tutors use. As an Organic Chemistry peer tutor, this technique, although it required more time, helped students solve problems with better understanding and retention of the information.
This past week a few of those who are eligible took a safety course in order to become C14 certified. Each lab is required to have someone on site who has this certificate of fitness and I am very excited to finally take the test and get my certification. With this certification, my research advisor said that I would be able to receive a copy of the keys to the lab. I am very excited to take on more responsibility. All in all, I am thrilled to continue working on the project I started last summer and further the research with substantial and reproducible results.