Integrating Genetics and Behavior: Unveiling the Genetic Landscape of Learning and Memory in Zebrafish through Breeding, Microinjections, and CRISPR-Cas9
Presenter Type
UNO Undergraduate Student
Major/Field of Study
Biology
Other
Molecular and Biomedical Biology
Advisor Information
Dr. Wong, Wong Lab
Location
CEC RM #201/205/209
Presentation Type
Poster
Poster Size
48x36
Start Date
22-3-2024 1:00 PM
End Date
22-3-2024 2:15 PM
Abstract
One key trait of survival is an animal’s ability to commit experiences to memory and learn from those situational outcomes. This process is complex including a cascade of cognitive processes, encompassing stimulus perception, information encoding, storage, and retrieval. Learning and memory along with other cognitive capabilities can differ between personality types, specifically the proactive and reactive stress coping styles observed in zebrafish. Qualitative differences in responses to stimuli and genetic composition have been studied between proactive and reactive stress coping styles. Whole brain RNA sequencing studies have indicated that reactive fish have higher amounts of npas4a and grm5 and proactive fish have higher amounts of comta and npy than their counterpart. suggest diverse social and ecological interpretations between individuals with alternative personality types.
To address this, our study uses the CRISPR-Cas9 system to insert mutations into the single-cell stage of the zebrafish embryos that will cause the deletion of these candidate genes in each respective stress-coping style. Our methodology includes breeding zebrafish with distinct stress coping styles, conducting microinjections to induce gene knockout, raising embryos to adulthood, and subsequently assessing learning and memory behavior.
Currently, over 60 zebrafish have been tested to find the most extreme candidates to breed for each stress-coping style. Breeding has been attempted 24 times, resulting in a total collection of roughly 600 eggs, with half of those being injected with a testing dye for practicing microinjection. Roughly 37% of the injected embryos survived and 41% of the uninjected embryos survived. We are testing different breeding strategies, such as breeder pairing ratios, methods of checking for eggs, and time tested for breeding.
Our novel breeding strategy exhibits promising results in achieving increased egg production and minimizing stress on breeding fish. The implications extend to our further research on microinjecting the embryos with the CRISPR-Cas9 system to remove targeted genes and raising the embryos to adulthood to see their influence on learning and memory in proactive and reactive stress coping styles.
Integrating Genetics and Behavior: Unveiling the Genetic Landscape of Learning and Memory in Zebrafish through Breeding, Microinjections, and CRISPR-Cas9
CEC RM #201/205/209
One key trait of survival is an animal’s ability to commit experiences to memory and learn from those situational outcomes. This process is complex including a cascade of cognitive processes, encompassing stimulus perception, information encoding, storage, and retrieval. Learning and memory along with other cognitive capabilities can differ between personality types, specifically the proactive and reactive stress coping styles observed in zebrafish. Qualitative differences in responses to stimuli and genetic composition have been studied between proactive and reactive stress coping styles. Whole brain RNA sequencing studies have indicated that reactive fish have higher amounts of npas4a and grm5 and proactive fish have higher amounts of comta and npy than their counterpart. suggest diverse social and ecological interpretations between individuals with alternative personality types.
To address this, our study uses the CRISPR-Cas9 system to insert mutations into the single-cell stage of the zebrafish embryos that will cause the deletion of these candidate genes in each respective stress-coping style. Our methodology includes breeding zebrafish with distinct stress coping styles, conducting microinjections to induce gene knockout, raising embryos to adulthood, and subsequently assessing learning and memory behavior.
Currently, over 60 zebrafish have been tested to find the most extreme candidates to breed for each stress-coping style. Breeding has been attempted 24 times, resulting in a total collection of roughly 600 eggs, with half of those being injected with a testing dye for practicing microinjection. Roughly 37% of the injected embryos survived and 41% of the uninjected embryos survived. We are testing different breeding strategies, such as breeder pairing ratios, methods of checking for eggs, and time tested for breeding.
Our novel breeding strategy exhibits promising results in achieving increased egg production and minimizing stress on breeding fish. The implications extend to our further research on microinjecting the embryos with the CRISPR-Cas9 system to remove targeted genes and raising the embryos to adulthood to see their influence on learning and memory in proactive and reactive stress coping styles.