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 (�) 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 �-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 �-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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