During my 2017 spring semester at UC I continued my research on Dr.

Tiwaris project in

the Gross Lab in the Department of Neurology at Cincinnati Childrens Hospital Medical Center.

The majority of my work during the semester was the continuation of older procedures in order

to collect more data in order to better prove the significance of our previous observation that a

pretreatment with miR-218 antagomir may provide a neuroprotective effect on mouse

hippocampal neurons after a kainic acid-induced seizures. Procedures that I performed included

genotyping of mice via tissue digestion, PCR, and gel electrophoresis, Fluoro-Jade B staining

(for the labeling of degenerating neurons), immunohistochemistry staining for Kv4.2 expression

(the voltage-gated potassium channel which is the focus of the project), and frozen brain

sectioning using a cryostat. Two key differences of this semester compared to previous semesters

are the comparing of intracerebroventricular vs direct hippocampal injections of miR-218 and the

new usage of mice whose hippocampi are injected with a lentivrus containing a plasmid which

encodes for the Kv4.2 gene in the CA1 region. The purpose of the lentiviral injections is to take a

different mechanistic approach at increasing Kv4.2 expression (this involves no interaction with

microRNA).

The photograph which I have chosen as an example of my work is a 4x magnified image

of a mouse brain which shows the lentiviral injection into the hippocampus. In this case, the

virus encodes for GFP, which fluoresces green when exposed to the correct wavelength of light.

The purpose of the GFP injections are to enable us to visualize how far the virus travels from the

injection site. In this case, it can be seen that the virus did not travel much at all, thus we are

prompted to optimize our procedure.