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Stress: Adolescence through Adulthood

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Anisha Kaldindi 

 I am in the research group of Dr. Gretchen Neigh in the Department of Physiology in the School of Medicine. I have been working on a project that involves looking at how chronic adolescent stress affects factors of the stress axis in adulthood.
We have developed a protocol in which our animals experience chronic stress during the adolescent period and then we collect the brain tissue in adulthood. Since we are interested in specific regions in the brain we use a machine called a cryostat that allows us to make very thin slices of the brain. There is a lot of literature on the anatomy of the brain in rats (our model system), so we are able to use a book called the Rat Brain Atlas, to look at the structural elements of the brain and determine where we can collect the tissue from the region we are interested in.

For us to understand how the stress factors are affected in adulthood, we use antibodies (a way to mark the factors we are interested in) and then analyze these images to see how groups that have experienced stress differ from groups that have not.
The protocol to do this had not been established in the lab previously, so I had the opportunity to work with people in our group to develop this protocol. This first involved optimizing the antibody concentrations. Since antibodies serve as a marker for the factors we are interested in, we need to make sure that there is no nonspecific binding.  To do this we tried varied concentrations of antibodies. We then used imaging software to determine if we were getting any nonspecific binding. This has been a long process and we have finally developed an optimal procedure.
Our next step was to determine how to image and analyze the data.
Currently, I am working on this while doing some final optimization of the protocol to determine where in the brain region of interest we need to focus on. At times this long drawn out process of troubleshooting has been frustrating but with the guidance of my mentors we have been able to develop a successful protocol. Through this project there have been days where a sample we run showed promise and it was these victories that have helped me sustain optimism and determination to finish this project.
-Anisha Kaldindi

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