Though this week was filled with dissections and cell culture, I spent a lot of my time running gels for western blots in the background.
In the beginning of the week, it was noticed that it’s necessary to sacrifice several cages with animals. This meant that they also had to be dissected afterword. This was a lengthy process as from each of them a skin sample was needed as well as their spleens, and lymph nodes. Lymph nodes are found all over the body and were taken, but the mesenteric ones are especially important for my experiment, as they are found attached to a particularly thin tissue around the intestines.
Each mouse took a considerable amount of time. In the background, however, I was continuing to run several gels; these were each aimed at determining the prominence of certain tight junction proteins in the small and large intestines. I also had to run a blot with actin as a control to insure that I can interpret the blots effectively.
After a literature review, we know which treatments given to the mice should theoretically cause greater intestinal permeability with to increased inflammation, and which should decrease gut permeability. According to studies, different concentrations of some tight junction protein account for either greater or lesser permeability in both the small and large intestines. Currently, we want to ensure that while we continue the treatment to regulate the gut’s permeability, it is indeed effective. Though I had reassuring results by this Friday with two proteins, I want to repeat it with a couple of other proteins next week.