Differentiation of Mammalian Vestibular Hair Cells from Conditionally Immortal, Postnatal Supporting Cells

Lawlor, Patrick; Marcotti, Walter; Rivolta, Marcelo N.; Kros, Corné J.; Holley, Matthew C.

doi:10.1523/jneurosci.19-21-09445.1999

Cited by 48 publications

(40 citation statements)

References 59 publications

(63 reference statements)

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“…Last, we suggest that replication of the physiological profile through which embryonic hair cells develop (Fig. 9) in candidate regenerated hair cells (Lawlor et al, 1999;Shou et al, 2003) will help validate their advancement to the enviable status of fully functional hair cell.…”

Section: Discussionmentioning

confidence: 86%

Developmental Acquisition of Voltage-Dependent Conductances and Sensory Signaling in Hair Cells of the Embryonic Mouse Inner Ear

Géléoc¹,

Risner²,

Holt³

2004

J. Neurosci.

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How and when sensory hair cells acquire the remarkable ability to detect and transmit mechanical information carried by sound and head movements has not been illuminated. Previously, we defined the onset of mechanotransduction in embryonic hair cells of mouse vestibular organs to be at approximately embryonic day 16 (E16). Here we examine the functional maturation of hair cells in intact sensory epithelia excised from the inner ears of embryonic mice. Hair cells were studied at stages between E14 and postnatal day 2 using the whole-cell, tight-seal recording technique. We tracked the developmental acquisition of four voltage-dependent conductances. We found a delayed rectifier potassium conductance that appeared as early as E14 and grew in amplitude over the subsequent prenatal week. Interestingly, we also found a low-voltage-activated potassium conductance present at E18, ϳ1 week earlier than reported previously. An inward rectifier conductance appeared at approximately E15 and doubled in size over the next few days. We also noted transient expression of a voltage-gated sodium conductance that peaked between E16 and E18 and then declined to near zero at birth. We propose that hair cells undergo a stereotyped developmental pattern of ion channel acquisition and that the precise pattern may underlie other developmental processes such as synaptogenesis and functional differentiation into type I and type II hair cells. In addition, we find that the developmental acquisition of basolateral conductances shapes the hair cell receptor potential and therefore comprises an important step in the signal cascade from mechanotransduction to neurotransmission.

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Section: Discussionmentioning

confidence: 86%

Developmental Acquisition of Voltage-Dependent Conductances and Sensory Signaling in Hair Cells of the Embryonic Mouse Inner Ear

Géléoc¹,

Risner²,

Holt³

2004

J. Neurosci.

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“…Oshima et al 2007;Savary et al 2007Savary et al , 2008Senn et al 2007). To identify differentiated cells, we used antibodies specific for the early hair cell markers myosin VIIa ) and parvalbumin 3 (Heller et al 2002), as well as antibodies to pan-cytokeratin, which label epithelial cells including supporting cells (Lawlor et al 1999;Li et al 2003;Zheng et al 1997). The morphology and the pattern of immunoreactivity for pan-cytokeratin after 5 days (Fig.…”

Section: Figmentioning

confidence: 99%

Stem/Progenitor Cells Derived from the Cochlear Sensory Epithelium Give Rise to Spheres with Distinct Morphologies and Features

Diensthuber

Oshima

Heller

2009

JARO

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Nonmammalian vertebrates regenerate lost sensory hair cells by means of asymmetric division of supporting cells. Inner ear or lateral line supporting cells in birds, amphibians, and fish consequently serve as bona fide stem cells resulting in high regenerative capacity of hair cell-bearing organs. Hair cell regeneration does not happen in the mammalian cochlea, but cells with proliferative capacity can be isolated from the neonatal cochlea. These cells have the ability to form clonal floating colonies, so-called spheres, when cultured in nonadherent conditions. We noticed that the sphere population derived from mouse cochlear sensory epithelium cells was heterogeneous, consisting of morphologically distinct sphere types, hereby classified as solid, transitional, and hollow. Cochlear sensory epithelium-derived stem/progenitor cells initially give rise to small solid spheres, which subsequently transition into hollow spheres, a change that is accompanied by epithelial differentiation of the majority of sphere cells. Only solid spheres, and to a lesser extent, transitional spheres, appeared to harbor self-renewing stem cells, whereas hollow spheres could not be consistently propagated. Solid spheres contained significantly more rapidly cycling Pax-2-expressing presumptive otic progenitor cells than hollow spheres. Islet-1, which becomes upregulated in nascent sensory patches, was also more abundant in solid than in hollow spheres. Likewise, hair cell-like cells, characterized by the expression of multiple hair cell markers, differentiated in significantly higher numbers in cell populations derived from solid spheres. We conclude that cochlear sensory epithelium cell populations initially give rise to small solid spheres that have self-renewing capacity before they subsequently convert into hollow spheres, a process that is accompanied by loss of stemness and reduced ability to spontaneously give rise to hair celllike cells. Solid spheres might, therefore, represent the most suitable sphere type for cell-based assays or animal model transplantation studies aimed at development of cell replacement therapies.

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“…Similarly, it is unlikely that screens utilizing in vitro preparations of utricles or cochleae from larger animals will be routinely adopted, as thousands of animals would be required to screen even mediumsized compound libraries. Hair cell lines (Rivolta et al 1998;Lawlor et al 1999) provide an alternative model that is more compatible for screening; however, to date, no screen of ototoxicity in the few lines available has been reported. Furthermore, the use of hair cell lines is controversial because these cells have been selected to survive and thus may not be an ideal model to screen for hair cell death.…”

Section: Alternative Approaches To Screeningmentioning

confidence: 99%

Using the Zebrafish Lateral Line to Screen for Ototoxicity

Chiu

Cunningham

Raible

et al. 2008

JARO

181

134

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The zebrafish is a valuable model for studying hair cell development, structure, genetics, and behavior. Zebrafish and other aquatic vertebrates have hair cells on their body surface organized into a sensory system called the lateral line. These hair cells are highly accessible and easily visualized using fluorescent dyes. Morphological and functional similarities to mammalian hair cells of the inner ear make the zebrafish a powerful preparation for studying hair cell toxicity. The ototoxic potential of drugs has historically been uncovered by anecdotal reports that have led to more formal investigation. Currently, no standard screen for ototoxicity exists in drug development. Thus, for the vast majority of Food and Drug Association (FDA)-approved drugs, the ototoxic potential remains unknown. In this study, we used 5-day-old zebrafish larvae to screen a library of 1,040 FDA-approved drugs and bioactives (NINDS Custom Collection II) for ototoxic effects in hair cells of the lateral line. Hair cell nuclei were selectively labeled using a fluorescent vital dye. For the initial screen, fish were exposed to drugs from the library at a 100-μM concentration for 1 h in 96-well tissue culture plates. Hair cell viability was assessed in vivo using fluorescence microscopy. One thousand forty drugs were rapidly screened for ototoxic effects. Seven known ototoxic drugs included in the library, including neomycin and cisplatin, were positively identified using these methods, as proof of concept.Fourteen compounds without previously known ototoxicity were discovered to be selectively toxic to hair cells. Dose-response curves for all 21 ototoxic compounds were determined by quantifying hair cell survival as a function of drug concentration. Dose-response relationships in the mammalian inner ear for two of the compounds without known ototoxicity, pentamidine isethionate and propantheline bromide, were then examined using in vitro preparations of the adult mouse utricle. Significant dose-dependent hair cell loss in the mouse utricle was demonstrated for both compounds. This study represents an important step in validating the use of the zebrafish lateral line as a screening tool for the identification of potentially ototoxic drugs.

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Differentiation of Mammalian Vestibular Hair Cells from Conditionally Immortal, Postnatal Supporting Cells

Cited by 48 publications

References 59 publications

Developmental Acquisition of Voltage-Dependent Conductances and Sensory Signaling in Hair Cells of the Embryonic Mouse Inner Ear

Developmental Acquisition of Voltage-Dependent Conductances and Sensory Signaling in Hair Cells of the Embryonic Mouse Inner Ear

Stem/Progenitor Cells Derived from the Cochlear Sensory Epithelium Give Rise to Spheres with Distinct Morphologies and Features

Using the Zebrafish Lateral Line to Screen for Ototoxicity

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