Wednesday, November 25, 2015

URSP Student Johanna Mingos Researches the Effect of Zinc on BDNF in 9-month-old Transgenic Mice Modeling Alzheimer’s Disease

I have always been fascinated by viruses, bacteria, and infectious diseases, and my microbiology labs have reinforced my passion to work in a research lab after graduation. During my junior year, I became an undergraduate research assistant under the direction of Dr. Jane Flinn of the Cognitive and Behavioral Neuroscience department. Dr. Flinn’s lab examines the role that metals play in Alzheimer’s disease, and I quickly learned that the skills and techniques I had practiced in my Biology labs easily translated to the field of psychology. Assisting a doctoral candidate with Western Blot analysis to examine inflammatory proteins in mice, I learned new laboratory techniques, experimental design, and data analysis. I am continuing this research under USRP as I examine the relationship between zinc and brain-derived neurotrophic factor (BDNF) protein in mice modeling Alzheimer’s disease, and I am looking forward to acquiring additional laboratory skills that I will be able to use across a range of disciplines.

Each week, I conduct a Western Blot analysis on approximately sixteen samples. After performing an assay to determine sample concentrations, I transfer the proteins to imaging membranes via gel electrophoresis. The membranes are incubated overnight with a primary antibody, followed by a secondary antibody and chemiluminescent substrate the next day. The membranes are then exposed using specialized software, and the resulting digital images are analyzed to determine the amount of BDNF present in each animal. The membranes are then re-probed for additional proteins, allowing me to carry out my USRP project while continuing my research assistant duties.

Occasionally, the discoveries made in the lab arise from a lack of experimental results; there can be surprises as we learn about new techniques or more efficient protocols. For example, when the recommended dilution of a specific antibody did not produce membrane images for either control or experimental animals, we used literature searches and additional experimentation to troubleshoot the issue. We determined the correct dilution necessary for our specific protocol, and we were able to obtain viable results in subsequent imaging.