Month/Year of Graduation

5-2025

Degree Name

Bachelor of Science (B.S.)

Department

Biology

Abstract

Reactive oxygen species are byproducts of cellular metabolism that, when not efficiently neutralized, can damage DNA, proteins, and lipids. Superoxide dismutase 1 (SOD1) is an antioxidant enzyme that converts superoxide radicals into less harmful molecules. While its molecular function is well documented, the role of SOD1 in shaping stress-related behavior remains underexplored. This project aimed to establish foundational tools for studying the behavioral impact of SOD1 knockout across proactive and reactive stress-coping styles in zebrafish.

To support future gene-editing experiments, microinjection and breeding protocols were refined across wildtype AB, proactive, and reactive zebrafish lines. No embryos were injected with CRISPR-Cas9 reagents due to limited availability of single-cell stage embryos, particularly in proactive and reactive strains. A total of 233 breeding attempts were conducted to determine optimal spawning conditions. Critical variables included strain type, sex ratio, pre-isolation period, egg collection timing, tank design, submersion duration, and minimizing disturbance. The most effective setup involved a 2:1 female-to-male ratio, four-day pre-isolation, and a custom-designed breeding chamber left submerged and undisturbed until the second morning. Eggs were detected using a borescope.

Microinjection techniques were validated using phenol red dye, and embryo survival was tracked across all strains. A total of 315 reactive, 850 proactive, and 1,641 wildtype embryos were assessed. Survival was slightly lower in injected groups compared to uninjected controls, but injection viability was confirmed. This study establishes a strain-sensitive breeding and injection protocol that lays essential groundwork for investigating how oxidative stress and behavioral phenotype interact to shape stress responses.

Download

Share

COinS