scholarly journals A Xenopus neuromast bioassay for chemical ototoxicity

2022 ◽  
Author(s):  
V. Bleu Knight ◽  
Amanda R. Luna ◽  
Elba Serrano
Keyword(s):  
Hair Cell ◽  
Inner Ear ◽  
Lateral Line ◽  
Hair Cells ◽  
Experimental Manipulation ◽  
The Body ◽  
Therapeutic Drug ◽  
Sem Images ◽  
Irreversible Damage ◽  
Mechanosensory Hair Cells

Background: Ototoxic chemicals can impair the senses of hearing and balance in mammals through irreversible damage to the mechanosensory bundles of inner ear hair cells. Fish and amphibians are useful models for investigating ototoxicity because their inner ear hair cells, like those of mammals, are susceptible to damage by ototoxins. Moreover, amphibian mechanosensation is augmented by a lateral line organ on the body surface that comprises external mechanosensory hair cells. The lateral line hair cells are arranged in clusters (neuromasts) and are structurally and functionally similar to inner ear hair cells, but are more accessible for experimental manipulation. Herein, we implemented neuromasts of the amphibian (Xenopus) lateral line as an organ system for evaluating the effects of ototoxic chemicals, such as antibiotics, on mechanosensory hair cell bundles. Methods: We examined the ultrastructure of larval Xenopus laevis neuromasts with scanning electron microscopy (SEM) after larvae were continuously exposed to ototoxic aminoglycoside antibiotics at sub-lethal concentrations (gentamicin; streptomycin; neomycin) for 72 hours. Results: SEM images demonstrated that 72 hours of exposure to antibiotic concentrations greater than 25 μM reduced the hair cell bundle number in lateral line neuromasts. Conclusion: Therapeutic drug studies will benefit from the incorporation of bioassay strategies that evaluate ototoxicity across multiple species including genera of amphibian origin such as Xenopus. Our outcomes support the use of the Xenopus lateral line for identification of potential ototoxic chemicals and suggest that Xenopus neuromast hair cell bundles can withstand antibiotic exposure. The Xenopus bioassay presented here can be incorporated into drug discovery methodology as a high-resolution phenotypic screen for ototoxic effects.

scholarly journals The cell biology of hearing

2010 ◽  
Vol 190 (1) ◽  
pp. 9-20 ◽  
Author(s):  
Martin Schwander ◽  
Bechara Kachar ◽  
Ulrich Müller
Keyword(s):  
Hair Cell ◽  
Inner Ear ◽  
Hair Cells ◽  
Cell Biology ◽  
Cell Development ◽  
Biological Mechanisms ◽  
Electrical Signals ◽  
Mechanosensory Hair Cells

Mammals have an astonishing ability to sense and discriminate sounds of different frequencies and intensities. Fundamental for this process are mechanosensory hair cells in the inner ear that convert sound-induced vibrations into electrical signals. The study of genes that are linked to deafness has provided insights into the cell biological mechanisms that control hair cell development and their function as mechanosensors.

scholarly journals Macrophages Respond Rapidly to Ototoxic Injury of Lateral Line Hair Cells but Are Not Required for Hair Cell Regeneration

2021 ◽  
Vol 14 ◽  
Author(s):  
Mark E. Warchol ◽  
Angela Schrader ◽  
Lavinia Sheets
Keyword(s):  
Hair Cell ◽  
Inner Ear ◽  
Lateral Line ◽  
Hair Cells ◽  
Control Fish ◽  
Cell Regeneration ◽  
Cellular Interactions ◽  
Hair Cell Regeneration ◽  
Genetic Ablation ◽  
Larval Zebrafish

The sensory organs of the inner ear contain resident populations of macrophages, which are recruited to sites of cellular injury. Such macrophages are known to phagocytose the debris of dying cells but the full role of macrophages in otic pathology is not understood. Lateral line neuromasts of zebrafish contain hair cells that are nearly identical to those in the inner ear, and the optical clarity of larval zebrafish permits direct imaging of cellular interactions. In this study, we used larval zebrafish to characterize the response of macrophages to ototoxic injury of lateral line hair cells. Macrophages migrated into neuromasts within 20 min of exposure to the ototoxic antibiotic neomycin. The number of macrophages in the near vicinity of injured neuromasts was similar to that observed near uninjured neuromasts, suggesting that this early inflammatory response was mediated by “local” macrophages. Upon entering injured neuromasts, macrophages actively phagocytosed hair cell debris. The injury-evoked migration of macrophages was significantly reduced by inhibition of Src-family kinases. Using chemical-genetic ablation of macrophages before the ototoxic injury, we also examined whether macrophages were essential for the initiation of hair cell regeneration. Results revealed only minor differences in hair cell recovery in macrophage-depleted vs. control fish, suggesting that macrophages are not essential for the regeneration of lateral line hair cells.

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 Macrophages respond rapidly to ototoxic injury of lateral line hair cells but are not required for hair cell regeneration

2020 ◽  
Author(s):  
Mark E. Warchol ◽  
Angela Schrader ◽  
Lavinia Sheets
Keyword(s):  
Hair Cell ◽  
Inner Ear ◽  
Lateral Line ◽  
Hair Cells ◽  
Control Fish ◽  
Cell Regeneration ◽  
Cellular Interactions ◽  
Hair Cell Regeneration ◽  
Genetic Ablation ◽  
Larval Zebrafish

AbstractThe sensory organs of the inner ear contain resident populations of macrophages, which are recruited to sites of cellular injury. Such macrophages are known to phagocytose the debris of dying cells but the full role of macrophages in otic pathology is not understood. Lateral line neuromasts of zebrafish contain hair cells similar to those in the inner ear, and the optical clarity of larval zebrafish permits direct imaging of cellular interactions. In this study, we used larval zebrafish to characterize the response of macrophages to ototoxic injury of lateral line hair cells. Macrophages migrated into neuromasts within 20 min of exposure to the ototoxic antibiotic neomycin. The number of macrophages in close proximity of injured neuromasts was similar to that observed near uninjured neuromasts, suggesting that this early inflammatory response was mediated by ‘local’ macrophages. Upon entering injured neuromasts, macrophages actively phagocytosed hair cell debris. Such phagocytosis was significantly reduced by inhibiting Src-family kinases. Using chemical-genetic ablation of macrophages prior to ototoxic injury, we also examined whether macrophages were essential for the initiation of hair cell regeneration after neomycin exposure. Results revealed only minor differences in hair cell recovery in macrophage-depleted vs. control fish, suggesting that macrophages are not essential for the regeneration of lateral line hair cells.

scholarly journals Distinct progenitor populations mediate regeneration in the zebrafish lateral line

eLife ◽  
2019 ◽  
Vol 8 ◽  
Author(s):  
Eric D Thomas ◽  
David W Raible
Keyword(s):  
Hair Cell ◽  
Lateral Line ◽  
Hair Cells ◽  
Cell Damage ◽  
Cell Populations ◽  
Support Cell ◽  
Cell Ablation ◽  
Hair Cell Damage ◽  
Transgenic Lines ◽  
Mechanosensory Hair Cells

Mechanosensory hair cells of the zebrafish lateral line regenerate rapidly following damage. These renewed hair cells arise from the proliferation of surrounding support cells, which undergo symmetric division to produce two hair cell daughters. Given the continued regenerative capacity of the lateral line, support cells presumably have the ability to replenish themselves. Utilizing novel transgenic lines, we identified support cell populations with distinct progenitor identities. These populations show differences in their ability to generate new hair cells during homeostasis and regeneration. Targeted ablation of support cells reduced the number of regenerated hair cells. Furthermore, progenitors regenerated after targeted support cell ablation in the absence of hair cell damage. We also determined that distinct support cell populations are independently regulated by Notch signaling. The existence of independent progenitor populations could provide flexibility for the continued generation of new hair cells under a variety of conditions throughout the life of the animal.

scholarly journals Distinct progenitor populations mediate regeneration in the zebrafish lateral line

2018 ◽  
Author(s):  
Eric D. Thomas ◽  
David W. Raible
Keyword(s):  
Hair Cell ◽  
Lateral Line ◽  
Hair Cells ◽  
Cell Damage ◽  
Cell Populations ◽  
Support Cell ◽  
Cell Ablation ◽  
Hair Cell Damage ◽  
Transgenic Lines ◽  
Mechanosensory Hair Cells

ABSTRACTMechanosensory hair cells of the zebrafish lateral line regenerate rapidly following damage. These renewed hair cells arise from the proliferation of surrounding support cells, which undergo symmetric division to produce two hair cell daughters. Given the continued regenerative capacity of the lateral line, support cells presumably have the ability to replenish themselves. Utilizing novel transgenic lines, we identified support cell populations with distinct progenitor identities. These populations show differences in their ability to generate new hair cells during homeostasis and regeneration. Targeted ablation of support cells reduced the number of regenerated hair cells. Furthermore, progenitors regenerated after targeted support cell ablation in the absence of hair cell damage. We also determined that distinct support cell populations are independently regulated by Notch signaling. The existence of independent progenitor populations could provide flexibility for the continued generation of new hair cells under a variety of conditions throughout the life of the animal.

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.

The fine structure of the lateral-line sense organs of dogfish

1971 ◽  
Vol 179 (1055) ◽  
pp. 157-169 ◽  
Keyword(s):  
Hair Cell ◽  
Lateral Line ◽  
Hair Cells ◽  
Sensory Cell ◽  
Sensory Nerve ◽  
The Body ◽  
Apical Surface ◽  
Sense Organs ◽  
Supporting Cells ◽  
Efferent Nerve

The sense organs of the body lateral-line canals of Scyliorhinus were examined with the electron microscope and shown to consist of supporting cells and two kinds of sensory cell. One type of sensory cell has the well-known structure of hair cells, bearing on its apical surface a group of stereocilia (6 to 25) associated with a single kinocilium. Each hair cell is innervated by a sensory nerve fibre and some also receive an efferent nerve supply. The second kind of sensory cell is similar in appearance, but differs at the apex in containing many vacuoles and in lacking stereocilia. There are many long microvilli and a single cilium which arises from a shallow pit. The internal structure of this cilium is variable, with the number of tubules in the outer ring ranging between 7 and 9 and with the inner pair consisting of double elements. This type of sensory cell is innervated by sensory nerve fibres and possibly by efferent fibres. The situation of the kinocilium of a hair cell in relation to the stereocilia is more variable than has been described in other hair cells while the cilium of the second sensory cell appears to bear no special relation to the microvilli. The accessory cells of the neuromast include basal and peripheral supporting cells, many of which produce a secretion, and a large secretory cell which is found at intervals at the edge of the organ. This cell has a convoluted surface and is full of vesicles.

Hes1 is a negative regulator of inner ear hair cell differentiation

Development ◽  
2000 ◽  
Vol 127 (21) ◽  
pp. 4551-4560 ◽  
Author(s):  
J.L. Zheng ◽  
J. Shou ◽  
F. Guillemot ◽  
R. Kageyama ◽  
W.Q. Gao
Keyword(s):  
Cell Differentiation ◽  
Hair Cell ◽  
Inner Ear ◽  
Cell Fate ◽  
Hair Cells ◽  
Outer Hair Cells ◽  
Negative Regulator ◽  
Pcr Analysis ◽  
Loss Of Function ◽  
Hair Cell Differentiation

Hair cell fate determination in the inner ear has been shown to be controlled by specific genes. Recent loss-of-function and gain-of-function experiments have demonstrated that Math1, a mouse homolog of the Drosophila gene atonal, is essential for the production of hair cells. To identify genes that may interact with Math1 and inhibit hair cell differentiation, we have focused on Hes1, a mammalian hairy and enhancer of split homolog, which is a negative regulator of neurogenesis. We report here that targeted deletion of Hes1 leads to formation of supernumerary hair cells in the cochlea and utricle of the inner ear. RT-PCR analysis shows that Hes1 is expressed in inner ear during hair cell differentiation and its expression is maintained in adulthood. In situ hybridization with late embryonic inner ear tissue reveals that Hes1 is expressed in supporting cells, but not hair cells, of the vestibular sensory epithelium. In the cochlea, Hes1 is selectively expressed in the greater epithelial ridge and lesser epithelial ridge regions which are adjacent to inner and outer hair cells. Co-transfection experiments in postnatal rat explant cultures show that overexpression of Hes1 prevents hair cell differentiation induced by Math1. Therefore Hes1 can negatively regulate hair cell differentiation by antagonizing Math1. These results suggest that a balance between Math1 and negative regulators such as Hes1 is crucial for the production of an appropriate number of inner ear hair cells.

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