University of Iowa Department of Anatomy and Cell Biology
August 1, 2013
Week 7 was wrapping up my experiment as much as I could. I finished running qPCRs using the cDNA from a NFN trial. After that, I compiled that data to determine if my results were significant or not. I was extremely pleased to find out that NFN significantly decreases TH1 and TH2. Yay!! However, the next thing we became concerned about was if NFN was affecting dopaminergic neurons specifically or if the whole brain was just fried. I then ran a few qPCRs looking at Brain Derived Neurotrophic Factor (BDNF) and Glutamate Decarboxylase (GAD). BDNF would allow us to assess the brain overall, and GAD would allow us to look at neurons that are glutamate specific. By the end of the week, I had analyzed that NFN does not significantly affect either of these expressions. Our results suggest that NFN affects dopaminergic neurons specifically.
The other thing that I’ve been working on is my TH2 probe. I haven’t had any success getting good expression with my embryos which is a little disappointing. I’ve adjusted the protocol which I was hoping would help. The embryos now overdevelop really easily, but putting them in 100% methanol is supposed to help clear up background, so maybe by Monday of next week, they will look good. It would be nice if I could have pictures for my poster!
I also injected the RNA I synthesized from the gene I cloned (vmat2) into embryos. I’m really excited that I had a higher survival rate this time around, and I think I understand what I’m doing wrong. Hopefully, I will be able to be successful if I need to inject again.
Barbara and I set up another experiment with pargyline, so we can assess their behavior next week.
Otherwise, I’ve just been working on my poster. Right now, I’m working on the wording for the introduction, methods, and conclusion. I’m planning on having that portion done by Friday.
Major: Biochemistry & Molecular Biology. Hometown:Ankeny, Iowa.