TY - JOUR
T1 - Early development of hearing in zebrafish
AU - Lu, Zhongmin
AU - Desmidt, Alexandra A.
N1 - Funding Information:
This work was supported by the National Institute of Deafness and Other Communicative Disorders of the National Institutes of Health (R21DC009879), University of Miami Provost Research Award, and College of Arts and Sciences Gabelli Fellowship. We are grateful to Dr. Tatjana Piotrowsk’s lab for providing us the SqET4 transgenic zebrafish. We would like to thank Dr. A. James Hudspeth for his advice during our development of the microphonic recording method. Thanks also to Ricardo Cepeda for excellent fish care; undergraduates Max Frye, Brittany Tyson, Adam Polinsky, Aleks Vantraub, Steven Basart, and Louis Cai for their help in the various aspects of this project; and Lilliann Zamora for her comments on this manuscript.
PY - 2013/8
Y1 - 2013/8
N2 - The zebrafish (Danio rerio) has become a valuable vertebrate model for human hearing and balance disorders because it combines powerful genetics, excellent embryology, and exceptional in vivo visualization in one organism. In this study, we investigated auditory function of zebrafish at early developmental stages using the microphonic potential method. This is the first study to report ontogeny of response of hair cells in any fish during the first week post fertilization. The right ear of each zebrafish embedded in agarose was linearly stimulated with a glass probe that was driven by a calibrated piezoelectric actuator. Using beveled micropipettes filled with standard fish saline, extracellular microphonic potentials were recorded from hair cells in the inner ear of zebrafish embryos or larvae in response to 20, 50, 100, and 200-Hz stimulation. Saccular hair cells expressing green fluorescent protein of the transgenic zebrafish from 2 to 7 days post fertilization (dpf) were visualized and quantified using confocal microscopy. The otic vesicles' areas, otoliths' areas, and saccular hair cell count and density increased linearly with age and standard body length. Microphonic responses increased monotonically with stimulus intensity, stimulus frequency, and age of zebrafish. Microphonic threshold at 200 Hz gradually decreased with zebrafish age. The increases in microphonic response and sensitivity correlate with the increases in number and density of hair cells in the saccule. These results enhance our knowledge of early development of auditory function in zebrafish and provide the control data that can be used to evaluate hearing of young zebrafish morphants or mutants.
AB - The zebrafish (Danio rerio) has become a valuable vertebrate model for human hearing and balance disorders because it combines powerful genetics, excellent embryology, and exceptional in vivo visualization in one organism. In this study, we investigated auditory function of zebrafish at early developmental stages using the microphonic potential method. This is the first study to report ontogeny of response of hair cells in any fish during the first week post fertilization. The right ear of each zebrafish embedded in agarose was linearly stimulated with a glass probe that was driven by a calibrated piezoelectric actuator. Using beveled micropipettes filled with standard fish saline, extracellular microphonic potentials were recorded from hair cells in the inner ear of zebrafish embryos or larvae in response to 20, 50, 100, and 200-Hz stimulation. Saccular hair cells expressing green fluorescent protein of the transgenic zebrafish from 2 to 7 days post fertilization (dpf) were visualized and quantified using confocal microscopy. The otic vesicles' areas, otoliths' areas, and saccular hair cell count and density increased linearly with age and standard body length. Microphonic responses increased monotonically with stimulus intensity, stimulus frequency, and age of zebrafish. Microphonic threshold at 200 Hz gradually decreased with zebrafish age. The increases in microphonic response and sensitivity correlate with the increases in number and density of hair cells in the saccule. These results enhance our knowledge of early development of auditory function in zebrafish and provide the control data that can be used to evaluate hearing of young zebrafish morphants or mutants.
KW - hair cell
KW - inner ear
KW - otic vesicle
KW - otolith organ
KW - saccule
KW - utricle
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U2 - 10.1007/s10162-013-0386-z
DO - 10.1007/s10162-013-0386-z
M3 - Article
C2 - 23575600
AN - SCOPUS:84880282120
VL - 14
SP - 509
EP - 521
JO - JARO - Journal of the Association for Research in Otolaryngology
JF - JARO - Journal of the Association for Research in Otolaryngology
SN - 1525-3961
IS - 4
ER -