News at Buckley

Zebrafish & Cortisol Inheritance

Ana Paula G.
My Applied Science project investigates the epigenetic aspects of stress in zebrafish (Danio rerio) by measuring cortisol levels through ELISA test.
This is important because if the offspring are born stressed, then I can start to find a way to destress the fish. I will be doing this by stressing the fish out and then mating them. To mate the fish, I will put one female in one side of the mating tank and a male on the other side of the mating tank. For a few hours, I will leave them there with the clear divider between them as the stressor to prevent mating. After a timed stress period, I will release the divider to allow them to mate. Then I will wait for the embryos to drop from the female so I can test them for their cortisol levels by using an ELISA Kit which helps me measure the cortisol in their feces. I will collect the embryos with a plate where the embryos will fall and land in holes, leading it easier for me to collect them. I can also measure cortisol levels by the cortisol released into the water through their gills. Embryos will be kept and grown and I will follow their growth and cortisol levels until 14 days post-fertilization. ​Cortisol levels will be tested from the stressed fish feces and aquarium water. The control is having them mate without a bar between them yet stressing them when they do have a bar between them. I hypothesize that stress will be genetic and the fish will be born more stressed than their parents because of how the proteins inside their bodies will be changed throughout their growth.

Some results I have come across are that my embryos are being born deformed. Though this sounds like bad news, this is actually helping verify my hypothesis. The deformity means that the embryos are being born pre-stressed, which means I am on the correct path for my project. The downside to these results are that none of the embryos have lived to the 14-day period, therefore, I have not been able to study the embryos closely. 
I have been doing this project since tenth grade, and this year I am adding more variables to my project. This year, I am, along with using regular zebrafish, using glow-in-the-dark zebrafish which have been injected with a chemical that makes them glow. I am using these fish to see if they have higher cortisol levels than the regular zebrafish and see if there are any differences with the embryos. I am also using the roundworm, C. elegans, to study their cortisol levels, compared to that of the zebrafish. 
Some accolades that I have received by doing this project are: 
  • Tenth grade LA Science Fair: third place award in my division of Biochemistry and Molecular Biology 
  • Eleventh grade Archer STEM Research Grant: I won $1,250 for my work

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