This Summer I had the privilege of interning under Dr. Alex Crawford at the Center for Molecular Medicine Norway. For this internship I got to live and work in Oslo, Norway for eight
weeks. My goal for the Summer was to screen medicinal plants and marine natural products, in collaboration with the University of Athens and University of South Florida, to discover compounds that may have anti-epileptic effects.

Using the model system zebrafish was an integral piece of screening compounds. Zebrafish larvae were used to find the maximal tolerance concentration that compounds could be
tested at, and larvae were used to see the effect the compounds had on convulsions. The first techniques I learned were the maintenance and breeding of zebrafish. Through the guidance of the lab technician, Alejandro, I learned the day to day maintenance of the adult zebrafish. This included multiple feedings per day of the thousands of zebrafish housed in the zebrafish room. The feedings consisted of not only dry food, but also artemia which are tiny shrimp. Artemia feeding was an enrichment for the zebrafish, as they had to hunt and catch the shrimp. The zebrafish tanks needed to be routinely cleaned, and water routinely checked, to ensure the fish were in an optimal environment. I also learned how to breed the adult zebrafish, as I had to regularly produce larvae for my experiment. To breed the zebrafish I had to first differ which fish were male and female, and sometimes these differences could be very subtle. Males and females were put into breeding tanks, which had a barrier to separate males and females. After being separated overnight the barrier was lifted in the morning and the fish would produce many fertilized eggs within just a couple hours. The eggs would then be collected and put into petri dishes and would grow into larvae, which I would use for experiments at four days post fertilization.

Once I knew the ins and outs of maintenance and breeding of zebrafish, I learned the techniques of the experiments I would be performing. Under PhD students, Parisa and Elham, I learned how to do maximal tolerance concentration and tracking experiments. The goal of the maximal tolerance concentration experiments was to find the highest concentration of the compounds that could be used on the fish, without toxicity. For MTC the compounds were solubilized in DMSO and diluted in E3 to the concentration which they would be tested at. Typically the first MTC was done at 100, 30, and 10 ug/mL and from there concentrations would be increased or decreased if needed. The diluted compound was put into a well that had a select
number of 4 day post fertilization larvae. The larvae were then left in the well overnight, and checked the next morning to find what the highest concentration of compound could be used without causing toxicity. The larvae that were in the highest concentration of compound that did not cause toxicity were then transferred to a tracking plate, which consisted of only one larva per well. The tracking plate was then put in the tracking machine which would track and measure the larvaes movement over a certain period of time. I would track the larvae for 30 minutes, and then add PTZ to each well. PTZ induced convulsions in the larvae. I would then track the larvae again for 30 minutes. To analyze this data I would normalize total distance to the DMSO group of each plate. Compounds that were a ‘hit’, had roughly the same total distance as the DMSO group before PTZ was added, but had a decreased total distance compared to the DMSO group after PTZ was added. To have a decreased total distance after PTZ was added, meant the compound decreased convulsive activity. I would then graph the data on GraphPad Prism, so the data could be easily presented.

Throughout my eight weeks I completed hundreds of screens, and produced some interesting results that will be further looked into for possible antiepileptic effect. I learned so much in my eight weeks from working with zebrafish, to learning new lab techniques, and even getting a glimpse into the world of drug discovery. Most importantly, this internship has solidified the career path I want to go down, a career doing research in the industry.

Working under Dr. Crawford at Center for Molecular Medicine Norway has been a once in lifetime experience that I will forever cherish. I got to meet so many amazing people from all around the world, got to learn new lab techniques, and got to experience living in Oslo! Sounds good!