Teaching Dose-response Relationships Through Aminoglycoside Block of Mechanotransduction Channels in Lateral Line Hair Cells of Larval Zebrafish

J Undergrad Neurosci Educ. 2018 Dec 15;17(1):A40-A49. eCollection 2018 Fall.


Here we introduce a novel set of laboratory exercises for teaching about hair cell structure and function and dose-response relationships via fluorescence microscopy. Through fluorescent labeling of lateral line hair cells, students assay aminoglycoside block of mechanoelectrical transduction (MET) channels in larval zebrafish. Students acquire and quantify images of hair cells fluorescently labeled with FM 1-43, which enters the hair cell through MET channels. Blocking FM 1-43 uptake with different concentrations of dihydrostreptomycin (DHS) results in dose-dependent reduction in hair-cell fluorescence. This method allows students to generate dose-response curves for the percent fluorescence reduction at different concentrations of DHS, which are then visualized to examine the blocking behavior of DHS using the Hill equation. Finally, students present their findings in lab reports structured as scientific papers. Together these laboratory exercises give students the opportunity to learn about hair cell mechanotransduction, pharmacological block of ion channels, and dose-dependent relationships including the Hill equation, while also exposing students to the zebrafish model organism, fluorescent labeling and microscopy, acquisition and analysis of images, and the presentation of experimental findings. These simple yet comprehensive techniques are appropriate for an undergraduate biology or neuroscience classroom laboratory.

Keywords: DHS; FM 1-43; MET channel; dihydrostreptomycin; dose-response curves; drug competition; fluorescence microscopy; hair cells; lateral line; mechanoelectrical transduction channel; zebrafish.