Publication Date

2019

Document Type

Honors Project

Degree Name

Bachelor of Arts

Department

Neuroscience

Advisors

Adam C. Hall

Keywords

General anesthesia, Zebrafish, Assay, GABA-A receptor, Delta-subunit

Abstract

General anesthetics have been widely used in clinical practice for over 170 years without a precise understanding of how the anesthetized state is generated. While the mechanisms of general anesthesia are not completely understood, recent studies have found the γ-aminobutyric acid type A receptor (GABAAR) to be an important target for general anesthetics. Our research focused on the delta (𝛿) subunit of GABAAR and the impacts it has on anesthetic sensitivity. Using the CRISPR/cas9 system, we generated two knockout (KO) zebrafish (Danio Rerio) lines with indel mutations (a 9bp insertion and 7bp deletion) on exon 6 of gabrd gene. In order to assess the impact of the GABAAR 𝛿-subunit KO on propofol anesthetic effects, we developed a novel dose response assay that effectively showed the dose-dependent anesthetic effects of propofol in wildtype (WT) larval zebrafish locomotion over time (immobility, WT EC50 = 1.25 µM propofol) and diminished response to noxious stimulus (hypnosis, WT EC50 between 1.75 µM and 2 µM propofol). With the establishment of an assay in wildtype larval zebrafish, future studies aim to use this assay to assess the gabrd mutants and determine if the 𝛿-subunit confers a level of anesthetic sensitivity.

Rights

©2019 Jamie Chan. Access limited to the Smith College community and other researchers while on campus. Smith College community members also may access from off-campus using a Smith College log-in. Other off-campus researchers may request a copy through Interlibrary Loan for personal use.

Language

English

Comments

51 pages : color illustrations. Includes bibliographical references (pages 45-47)

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