PUBLICATION
Vital Dye Uptake of YO-PRO-1 and DASPEI Depends Upon Mechanoelectrical Transduction Function in Zebrafish Hair Cells
- Authors
- Patterson, A.S., Dugdale, J., Koleilat, A., Krauss, A., Hernandez-Herrera, G.A., Wallace, J.G., Petree, C., Varshney, G.K., Schimmenti, L.A.
- ID
- ZDB-PUB-241022-18
- Date
- 2024
- Source
- Journal of the Association for Research in Otolaryngology : JARO 25(6): 531-543 (Journal)
- Registered Authors
- Schimmenti, Lisa A., Varshney, Gaurav
- Keywords
- cdh23, myo7aa, Gadolinium(iii) chloride, Mechanoelectrical transduction, Vital dyes, Zebrafish (Danio rerio)
- MeSH Terms
-
- Hair Cells, Auditory*/physiology
- Benzoxazoles
- Xanthenes*
- Quaternary Ammonium Compounds*/pharmacology
- Fluorescent Dyes
- Zebrafish*
- Mechanotransduction, Cellular*
- Animals
- Pyridinium Compounds
- Quinolinium Compounds*
- PubMed
- 39433714 Full text @ J. Assoc. Res. Otolaryngol.
Citation
Patterson, A.S., Dugdale, J., Koleilat, A., Krauss, A., Hernandez-Herrera, G.A., Wallace, J.G., Petree, C., Varshney, G.K., Schimmenti, L.A. (2024) Vital Dye Uptake of YO-PRO-1 and DASPEI Depends Upon Mechanoelectrical Transduction Function in Zebrafish Hair Cells. Journal of the Association for Research in Otolaryngology : JARO. 25(6):531-543.
Abstract
Purpose Vital dyes allow the visualization of cells in vivo without causing tissue damage, making them a useful tool for studying lateral line and inner ear hair cells in living zebrafish and other vertebrates. FM1-43, YO-PRO-1, and DASPEI are three vital dyes commonly used for hair cell visualization. While it has been established that FM1-43 enters hair cells of zebrafish and other organisms through the mechanoelectrical transduction (MET) channel, the mechanism of entry into hair cells for YO-PRO-1 and DASPEI has not been established despite widespread use. We hypothesize that YO-PRO-1 and DASPEI entry into zebrafish hair cells is MET channel uptake dependent similar to FM1-43.
Methods To test this hypothesis, we used both genetic and pharmacologic means to block MET channel function. Genetic based MET channel assays were conducted with two different mechanotransduction defective zebrafish lines, specifically the myo7aa-/- loss of function mutant tc320b (p.Y846X) and cdh23-/- loss of function mutant (c.570-571del). Pharmacologic assays were performed with Gadolinium(III) Chloride (Gad(III)), a compound that can temporarily block mechanotransduction activity.
Results Five-day post fertilization (5dpf) myo7aa-/- and cdh23-/- larvae incubated with FM1-43, YO-PRO-1, and DASPEI all showed nearly absent uptake of each vital dye. Treatment of wildtype zebrafish larvae with Gad(III) significantly reduces uptake of FM1-43, YO-PRO-1, and DASPEI vital dyes.
Conclusion These results indicate that YO-PRO-1 and DASPEI entry into zebrafish hair cells is MET channel dependent similar to FM1-43. This knowledge expands the repertoire of vital dyes that can be used to assess mechanotransduction and MET channel function in zebrafish and other vertebrate models of hair cell function.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping