Inner-ear abnormalities and their functional consequences in Belgian Waterslager canaries (Serinus canarius)

Recent reports of elevated auditory thresholds in canaries of the Belgian Waterslager strain have shown that this strain has an inherited auditory deficit in which absolute auditory thresholds at high frequencies (i.e. above 2.0 kHz) are as much as 40 dB less sensitive than the thresholds of mixed-breed canaries and those of other strains. The measurement of CAP audiograms showed that the hearing deficit is already present at the level of the auditory nerve (Gleich and Dooling, 1992). Here we show gross abnormalities in the anatomy of the basilar papilla of Belgian Waterslager canaries at the level of the hair cell. The extent of these abnormalities was correlated with the amount of hearing deficit as measured behaviorally

Parameters of Growth in the Embryonic and Neonatal Chick Basilar Papilla

The growth of the basilar papilla in the chick cochlear duct was studied utilizing light, scanning, and transmission electron microscopy. The ages of the cochleae investigated ranged from embryonic day 6 to post-hatching day 7. The changes in the length and width of the basilar papilla as well as the establishment of its spatula-like shape were correlated with the maturation of the hair cells\u27 apical surfaces and the changes in the cellular organization of the sensory epithelium. The histological reorganization of the distal hair cell nuclei was concomitant with the broadening of the distal region of the basilar papilla and occurred at a later stage than the reorganization of the proximal hair cell nuclei. Since the stereociliary bundles on all the hair cells are differentiated quite early, it appears that the delayed reorganization of the distal nuclei is associated with anatomical constraints on the cochlear duct, rather than a later differentiation of the distal sensory epithelium. A clear understanding of how growth of the cochlear duct influences both the distribution of hair cells on the basilar papilla\u27s surface and the cellular organization in the sensory epithelium is critical to future studies correlating ultrastructural development with functional maturation of the auditory system

Reorganization of the chick basilar papilla after acoustic trauma

The auditory epithelium in birds and mammals consists of a postmitotic population of hair cells and supporting cells. Unlike mammals, birds can regenerate their auditory epithelia after trauma. Recent evidence indicates that supporting cells undergo mitosis after acoustic trauma, suggesting that supporting cells may transdifferentiate into hair cells. The goals of this study were to (1) characterize the responses of hair cells and supporting cells to acoustic trauma, and (2) determine whether hair cell loss is a prerequisite for generation of new hair cells. Chicks were exposed to an octave-band noise and their inner ears assayed with fluorescence or scanning electron microscopy. In one area of the basilar papilla, defined as the center of the lesion, extensive hair cell degeneration occurred. Expanded supporting cells obliterated degenerating hair cells and invaded spaces normally occupied by hair cells. Aggregates of DNA were found within the basilar papilla, suggesting that hair cell death and disintegration may occur within the epithelium. The epithelial sheet appeared structurally confluent at all times examined. Supporting cells exhibited altered apical contour in distal regions of the basilar papilla, where hair cell damage was mild or inconspicuous. Four days after noise exposure, newly generated hair cells were found in the center of the lesion and in the distal areas, where no hair cell loss could be detected. The results suggest that supporting cells may play an important role in maintenance and repair of the traumatized basilar papilla and raise the possibility that production of new hair cells is not dependent on hair cell loss in the immediate vicinity.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/50055/1/903300408_ftp.pd

WDR1 colocalizes with ADF and actin in the normal and noise-damaged chick cochlea

Auditory hair cells of birds, unlike hair cells in the mammalian organ of Corti, can regenerate following sound-induced loss. We have identified several genes that are upregulated following such an insult. One gene, WDR1 , encodes the vertebrate homologue of actin-interacting protein 1, which interacts with actin depolymerization factor (ADF) to enhance the rate of actin filament cleavage. We examined WDR1 expression in the developing, mature, and noise-damaged chick cochlea by in situ hybridization and immunocytochemistry. In the mature cochlea, WDR1 mRNA was detected in hair cells, homogene cells, and cuboidal cells, all of which contain high levels of F-actin. In the developing inner ear, WDR1 mRNA was detected in homogene cells and cuboidal cells by embryonic day 7, in the undifferentiated sensory epithelium by day 9, and in hair cells at embryonic day 16. We also demonstrated colocalization of WDR1, ADF, and F-actin in all three cell types in the normal and noise-damaged cochlea. Immediately after acoustic overstimulation, WDR1 mRNA was seen in supporting cells. These cells contribute to the structural integrity of the basilar papilla, the maintenance of the ionic barrier at the reticular lamina, and the generation of new hair cells. These results indicate that one of the immediate responses of the supporting cell after noise exposure is to induce WDR1 gene expression and thus to increase the rate of actin filament turnover. These results suggest that WDR1 may play a role either in restoring cytoskeletal integrity in supporting cells or in a cell signaling pathway required for regeneration. J. Comp. Neurol. 448:399–409, 2002. © 2002 Wiley-Liss, Inc.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/34464/1/10265_ftp.pd

Dimethyl Sulfoxide (DMSO) Exacerbates Cisplatin-induced Sensory Hair Cell Death in Zebrafish (Danio rerio)

Inner ear sensory hair cells die following exposure to aminoglycoside antibiotics or chemotherapeutics like cisplatin, leading to permanent auditory and/or balance deficits in humans. Zebrafish (Danio rerio) are used to study drug-induced sensory hair cell death since their hair cells are similar in structure and function to those found in humans. We developed a cisplatin dose-response curve using a transgenic line of zebrafish that expresses membrane-targeted green fluorescent protein under the control of the Brn3c promoter/enhancer. Recently, several small molecule screens have been conducted using zebrafish to identify potential pharmacological agents that could be used to protect sensory hair cells in the presence of ototoxic drugs. Dimethyl sulfoxide (DMSO) is typically used as a solvent for many pharmacological agents in sensory hair cell cytotoxicity assays. Serendipitously, we found that DMSO potentiated the effects of cisplatin and killed more sensory hair cells than treatment with cisplatin alone. Yet, DMSO alone did not kill hair cells. We did not observe the synergistic effects of DMSO with the ototoxic aminoglycoside antibiotic neomycin. Cisplatin treatment with other commonly used organic solvents (i.e. ethanol, methanol, and polyethylene glycol 400) also did not result in increased cell death compared to cisplatin treatment alone. Thus, caution should be exercised when interpreting data generated from small molecule screens since many compounds are dissolved in DMSO.National Institutes of Health (U.S.) (DC010998)National Institutes of Health (U.S.) (NIH DC010231)Harvard College (1780- )Sarah Fuller Foundation for Little Deaf Childre

Class III myosins shape the auditory hair bundles by limiting microvilli and stereocilia growth.

International audienceThe precise architecture of hair bundles, the arrays of mechanosensitive microvilli-like stereocilia crowning the auditory hair cells, is essential to hearing. Myosin IIIa, defective in the late-onset deafness form DFNB30, has been proposed to transport espin-1 to the tips of stereocilia, thereby promoting their elongation. We show that Myo3a(-/-)Myo3b(-/-) mice lacking myosin IIIa and myosin IIIb are profoundly deaf, whereas Myo3a-cKO Myo3b(-/-) mice lacking myosin IIIb and losing myosin IIIa postnatally have normal hearing. Myo3a(-/-)Myo3b(-/-) cochlear hair bundles display robust mechanoelectrical transduction currents with normal kinetics but show severe embryonic abnormalities whose features rapidly change. These include abnormally tall and numerous microvilli or stereocilia, ungraded stereocilia bundles, and bundle rounding and closure. Surprisingly, espin-1 is properly targeted to Myo3a(-/-)Myo3b(-/-) stereocilia tips. Our results uncover the critical role that class III myosins play redundantly in hair-bundle morphogenesis; they unexpectedly limit the elongation of stereocilia and of subsequently regressing microvilli, thus contributing to the early hair bundle shaping

Cell cycle of transdifferentiating supporting cells in the basilar papilla

Mitosis of supporting cells has been shown to contribute to the cellular repopulation of the basilar papilla after acoustic trauma. In the present work we report data obtained with light and transmission electron microscopy after acoustic trauma in chicks. We report changes that occur in cell shape, surface morphology, intercellular junctions, nuclear shape and location, and cytoplasmic organization of supporting cells after trauma. The findings strongly suggest that supporting cells transdifferentiate and that the proliferative pattern is similar to interkinetic nuclear migration, as previously shown in the developing neural tube and basilar papilla. S-phase nuclei were positioned adjacent to the basement membrane, suggesting that interaction with the extracellular matrix may occur during the cell cycle. Supporting cells divided with the long axis of the spindle parallel to the reticular lamina and displayed no signs of intercellular communication during mitosis. This suggested to us that the fate of the progeny cells is determined prior to mitosis and that the progeny may be of identical phenotypic fate. Dividing cells had a smooth apical surface. The smooth surface may provide a marker to help identify dividing cells with scanning electron microscope analysis.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/31271/1/0000177.pd

Return of auditory function following structural regeneration after acoustic trauma: Behavioral measures from quail,

After measuring baseline behavioral audiograms, three of four behaviorally trained quail and fifteen untrained cohorts were exposed to a 1.5-kHz octave-band noise at 116-dB SPL for 4 h. The trained birds were tested daily following the exposure and showed a steady recovery of absolute sensitivity with a return to normal absolute thresholds by post-exposure days 8-10. Thirteen untrained cohorts were sacrificed after various survival times to evaluate the structural condition of the car. The cohorts all showed regeneration of sensory cells similar to that seen in chicks. The effects of repeated acoustic trauma on recovery of sensitivity were evaluated by re-exposing the three trained birds and two untrained cohorts 106 days after the first exposure. One of the trained birds was exposed a third time, 113 days following the second exposure. The findings demonstrate that, following acoustic trauma, normal sensitivity returns prior to complete structural regeneration of the sensory epithelium and that repeated acoustic trauma may increase the time course of recovery of normal hearing sensitivity.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/31335/1/0000245.pd

Molecular regulation of auditory hair cell death and approaches to protect sensory receptor cells and/or stimulate repair following acoustic trauma

International audienceLoss of auditory sensory hair cells (HCs) is the most common cause of hearing loss. This review addresses the signaling pathways that are involved in the programmed and necrotic cell death of auditory HCs that occur in response to ototoxic and traumatic stressor events. The roles of inflammatory processes, oxidative stress, mitochondrial damage, cell death receptors, members of the mitogen-activated protein kinase (MAPK) signal pathway and pro- and anti-cell death members of the Bcl-2 family are explored. The molecular interaction of these signal pathways that initiates the loss of auditory HCs following acoustic trauma is covered and possible therapeutic interventions that may protect these sensory HCs from loss via apoptotic or non-apoptotic cell death are explored

Early microfilament reorganization in injured auditory epithelia

Microfilaments (MFs) play an important role in wound healing and other regenerative events. The purpose of this study was to characterize changes in the distribution of MFs in traumatized auditory epithelia and compare these changes between avian (regenerating) and mammalian (nonregenerating) ears. Chicks and guinea pigs were acoustically overstimulated and their auditory epithelia analyzed using fluorescence microscopy with phalloidin as a MF-specific marker. Immediately or several hours after overstimulation, we observed a substantial reduction of MFs in stereocilia and the cuticular plate. The circumferential belt of MF which is associated with the adherens junctional complex was constricted in damaged hair cells (HCs) as early as 1 day after the exposure. Concomitant with the junctional constriction, the apical surface area of supporting cells was increased relative to normal, whereas the surface area of HCs was decreased. We conclude that changes in the amount and distribution of MFs which characterize early responses to acoustic damage are similar in avian (regenerating) and mammalian (nonregenerating) auditory epithelia. We hypothesize that changes in MF-mediated tensile forces trigger the process of tissue repair in auditory epithelia in response to insult. In mammals the reorganization of MFs may help maintain the integrity of the reticular lamina and thereby prevent further damage. In contrast, early changes in MFs in chicks may play a role in regulating regenerative tissue responses.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/30263/1/0000664.pd