scholarly journals In vivo and in vitro biophysical properties of hair cells from the lateral line and inner ear of developing and adult zebrafish

2014 ◽  
Vol 592 (10) ◽  
pp. 2041-2058 ◽  
Author(s):  
Jennifer Olt ◽  
Stuart L. Johnson ◽  
Walter Marcotti
Keyword(s):  
Inner Ear ◽  
Lateral Line ◽  
Hair Cells ◽  
Adult Zebrafish ◽  
Biophysical Properties

scholarly journals Vestibular and Auditory Hair Cell Regeneration Following Targeted Ablation of Hair Cells With Diphtheria Toxin in Zebrafish

2021 ◽  
Vol 15 ◽  
Author(s):  
Erin Jimenez ◽  
Claire C. Slevin ◽  
Luis Colón-Cruz ◽  
Shawn M. Burgess
Keyword(s):  
Hair Cell ◽  
Inner Ear ◽  
Lateral Line ◽  
Hair Cells ◽  
Diphtheria Toxin ◽  
Adult Zebrafish ◽  
Hair Cell Regeneration ◽  
Cell Ablation ◽  
Aquatic Vertebrates

Millions of Americans experience hearing or balance disorders due to loss of hair cells in the inner ear. The hair cells are mechanosensory receptors used in the auditory and vestibular organs of all vertebrates as well as the lateral line systems of aquatic vertebrates. In zebrafish and other non-mammalian vertebrates, hair cells turnover during homeostasis and regenerate completely after being destroyed or damaged by acoustic or chemical exposure. However, in mammals, destroying or damaging hair cells results in permanent impairments to hearing or balance. We sought an improved method for studying hair cell damage and regeneration in adult aquatic vertebrates by generating a transgenic zebrafish with the capacity for targeted and inducible hair cell ablation in vivo. This model expresses the human diphtheria toxin receptor (hDTR) gene under the control of the myo6b promoter, resulting in hDTR expressed only in hair cells. Cell ablation is achieved by an intraperitoneal injection of diphtheria toxin (DT) in adult zebrafish or DT dissolved in the water for larvae. In the lateral line of 5 days post fertilization (dpf) zebrafish, ablation of hair cells by DT treatment occurred within 2 days in a dose-dependent manner. Similarly, in adult utricles and saccules, a single intraperitoneal injection of 0.05 ng DT caused complete loss of hair cells in the utricle and saccule by 5 days post-injection. Full hair cell regeneration was observed for the lateral line and the inner ear tissues. This study introduces a new method for efficient conditional hair cell ablation in adult zebrafish inner ear sensory epithelia (utricles and saccules) and demonstrates that zebrafish hair cells will regenerate in vivo after this treatment.

scholarly journals Vestibular and auditory hair cell regeneration following targeted ablation of hair cells with diphtheria toxin in zebrafish

2021 ◽  
Author(s):  
Erin Jimenez ◽  
Claire C Slevin ◽  
Luis Colon Cruz ◽  
Shawn M Burgess
Keyword(s):  
Hair Cell ◽  
Inner Ear ◽  
Lateral Line ◽  
Hair Cells ◽  
Diphtheria Toxin ◽  
Adult Zebrafish ◽  
Hair Cell Regeneration ◽  
Cell Ablation ◽  
Aquatic Vertebrates

Millions of Americans experience hearing or balance disorders due to loss of hair cells in the inner ear. The hair cells are mechanosensory receptors used in the auditory and vestibular organs of all vertebrates as well as the lateral line systems of aquatic vertebrates. In zebrafish and other non-mammalian vertebrates, hair cells turn over during homeostasis and regenerate completely after being destroyed or damaged by acoustic or chemical exposure. However in mammals, destroying or damaging hair cells results in permanent impairments to hearing or balance. We sought an improved method for studying hair cell damage and regeneration in adult aquatic vertebrates by generating a transgenic zebrafish with the capacity for targeted and inducible hair cell ablation in vivo. This model expresses the human diphtheria toxin receptor (hDTR) gene under the control of the myo6b promoter, resulting in hDTR expressed only in hair cells. Cell ablation is achieved by an intraperitoneal injection of diphtheria toxin (DT) in adult zebrafish or DT dissolved in the water for larvae. In the lateral line of 5 dpf zebrafish, ablation of hair cells by DT treatment occurred within 2 days in a dose-dependent manner. Similarly, in adult utricles and saccules, a single intraperitoneal injection of 0.05 ng DT caused complete loss of hair cells in the utricle and saccule by 5 days post-injection. Full hair cell regeneration was observed for the lateral line and the inner ear tissues. This study introduces a new method for efficient conditional hair cell ablation in adult zebrafish inner ear sensory epithelia (utricles and saccules) and demonstrates that zebrafish hair cells will regenerate in vivo after this treatment.

Hair cell development in vivo and in vitro: Analysis by using a monoclonal antibody specific to hair cells in the chick inner ear

2002 ◽  
Vol 445 (2) ◽  
pp. 176-198
Author(s):  
Kenji Kondo ◽  
Hiroshi Sagara ◽  
Kazushige Hirosawa ◽  
Kimitaka Kaga ◽  
Satsuki Matsushima ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Hair Cell ◽  
Inner Ear ◽  
Hair Cells ◽  
Cell Development ◽  
Vitro Analysis ◽  
In Vitro Analysis

scholarly journals Gradients in the biophysical properties of neonatal auditory neurons align with synaptic contact position and the intensity coding map of inner hair cells

eLife ◽  
2020 ◽  
Vol 9 ◽  
Author(s):  
Alexander L Markowitz ◽  
Radha Kalluri
Keyword(s):  
Hair Cells ◽  
Sound Intensity ◽  
Spatial Gradient ◽  
Auditory Neuron ◽  
Biophysical Properties ◽  
Auditory Neurons ◽  
Successful Model ◽  
Intensity Coding

Sound intensity is encoded by auditory neuron subgroups that differ in thresholds and spontaneous rates. Whether variations in neuronal biophysics contributes to this functional diversity is unknown. Because intensity thresholds correlate with synaptic position on sensory hair cells, we combined patch clamping with fiber labeling in semi-intact cochlear preparations in neonatal rats from both sexes. The biophysical properties of auditory neurons vary in a striking spatial gradient with synaptic position. Neurons with high thresholds to injected currents contact hair cells at synaptic positions where neurons with high thresholds to sound-intensity are found in vivo. Alignment between in vitro and in vivo thresholds suggests that biophysical variability contributes to intensity coding. Biophysical gradients were evident at all ages examined, indicating that cell diversity emerges in early post-natal development and persists even after continued maturation. This stability enabled a remarkably successful model for predicting synaptic position based solely on biophysical properties.

scholarly journals Organotypic Cocultures of Human Pluripotent Stem Cell Derived-Neurons with Mammalian Inner Ear Hair Cells and Cochlear Nucleus Slices

2019 ◽  
Vol 2019 ◽  
pp. 1-14 ◽  
Author(s):  
Tomoko Hyakumura ◽  
Stuart McDougall ◽  
Sue Finch ◽  
Karina Needham ◽  
Mirella Dottori ◽  
...  
Keyword(s):  
Stem Cells ◽  
Stem Cell ◽  
Inner Ear ◽  
Hair Cells ◽  
Cochlear Nucleus ◽  
Cell Types ◽  
Quantitative Evidence ◽  
Brainstem Slice

Stem cells have been touted as a source of potential replacement neurons for inner ear degeneration for almost two decades now; yet to date, there are few studies describing the use of human pluripotent stem cells (hPSCs) for this purpose. If stem cell therapies are to be used clinically, it is critical to validate the usefulness of hPSC lines in vitro and in vivo. Here, we present the first quantitative evidence that differentiated hPSC-derived neurons that innervate both the inner ear hair cells and cochlear nucleus neurons in coculture, with significantly more new synaptic contacts formed on target cell types. Nascent contacts between stem cells and hair cells were immunopositive for both synapsin I and VGLUT1, closely resembling expression of these puncta in endogenous postnatal auditory neurons and control cocultures. When hPSCs were cocultured with cochlear nucleus brainstem slice, significantly greater numbers of VGLUT1 puncta were observed in comparison to slice alone. New VGLUT1 puncta in cocultures with cochlear nucleus slice were not significantly different in size, only in quantity. This experimentation describes new coculture models for assessing auditory regeneration using well-characterised hPSC-derived neurons and highlights useful methods to quantify the extent of innervation on different cell types in the inner ear and brainstem.

The Effect of Tgfß-2 on Inner Ear Development: Light and Electron Microscopy Observations

1997 ◽  
Vol 3 (S2) ◽  
pp. 173-174
Author(s):  
R. Friedman ◽  
N. Paradies ◽  
S. Wert ◽  
T. Doetschman ◽  
E.L. Cardell
Keyword(s):  
Inner Ear ◽  
Hair Cells ◽  
Transforming Growth Factor Beta ◽  
Transforming Growth Factor ◽  
Light And Electron Microscopy ◽  
Membranous Labyrinth ◽  
Ear Development ◽  
Inner Ear Development

Transforming growth factor beta (TGFß) genes are linked to a variety of developmental processes and are the subject of in vivo and in vitro transgene research studies. We are evaluating TGFß-2 effects on mouse inner ear development, with emphasis on the cochlear duct (CD), by comparing plastic sections of intact inner ears from developmental day (D) 16.5,18.5 and 19.5 littermates with wildtype (+/+), heterozygous (+/−) and mutant (−/−) TGFß-2 genotypes as determined by polymerase chain reaction analysis of tail digests. Auditory and vestibular organs of all D16.5 mice appear similar: membranous labyrinth epithelium varies from simple cuboidal/low columnar to pseudostratified/stratified columnar. Surrounding mesenchyme varies in cell density regionally, the most cellular mesenchyme underlies areas of sensory epithelium. Sparse mesenchymal cell distribution in the vestibule and basal CD indicates sites of perilymph channel formation. The spiral and vestibular ganglia and their unmyelinated fibers are prominent. Otoconia and hair cells are present in the utricle (U) and saccule (S) maculae; hair cells are less easily identifiable in the CD.

scholarly journals SoxC transcription factors are essential for the development of the inner ear

2015 ◽  
Vol 112 (45) ◽  
pp. 14066-14071 ◽  
Author(s):  
Ksenia Gnedeva ◽  
A. J. Hudspeth
Keyword(s):  
Transcription Factors ◽  
Hair Cell ◽  
Inner Ear ◽  
Hair Cells ◽  
Supporting Cell ◽  
Sensory Epithelia ◽  
Enhanced Expression ◽  
Mechanosensory Receptors

Hair cells, the mechanosensory receptors of the inner ear, underlie the senses of hearing and balance. Adult mammals cannot adequately replenish lost hair cells, whose loss often results in deafness or balance disorders. To determine the molecular basis of this deficiency, we investigated the development of a murine vestibular organ, the utricle. Here we show that two members of the SoxC family of transcription factors, Sox4 and Sox11, are down-regulated after the epoch of hair cell development. Conditional ablation of SoxC genes in vivo results in stunted sensory organs of the inner ear and loss of hair cells. Enhanced expression of SoxC genes in vitro conversely restores supporting cell proliferation and the production of new hair cells in adult sensory epithelia. These results imply that SoxC genes govern hair cell production and thus advance these genes as targets for the restoration of hearing and balance.

scholarly journals Bone marrow mesenchymal stem cells are progenitors in vitro for inner ear hair cells

2007 ◽  
Vol 34 (1) ◽  
pp. 59-68 ◽  
Author(s):  
Sang-Jun Jeon ◽  
Kazuo Oshima ◽  
Stefan Heller ◽  
Albert S.B. Edge
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Mesenchymal Stem Cells ◽  
Inner Ear ◽  
Hair Cells ◽  
Marrow Mesenchymal Stem Cells
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