RESUMEN
A screen for protein tyrosine phosphatases (PTPs) expressed in the chick inner ear yielded a high proportion of clones encoding an avian ortholog of protein tyrosine phosphatase receptor Q (Ptprq), a receptor-like PTP. Ptprq was first identified as a transcript upregulated in rat kidney in response to glomerular nephritis and has recently been shown to be active against inositol phospholipids. An antibody to the intracellular domain of Ptprq, anti-Ptprq, stains hair bundles in mice and chicks. In the chick ear, the distribution of Ptprq is almost identical to that of the 275 kDa hair-cell antigen (HCA), a component of hair-bundle shaft connectors recognized by a monoclonal antibody (mAb) that stains inner-ear hair bundles and kidney glomeruli. Furthermore, anti-Ptprq immunoblots a 275 kDa polypeptide immunoprecipitated by the anti-HCA mAb from the avian inner ear, indicating that the HCA and Ptprq are likely to be the same molecule. In two transgenic mouse strains with different mutations in Ptprq, anti-Ptprq immunoreactivity cannot be detected in the ear. Shaft connectors are absent from mutant vestibular hair bundles, but the stereocilia forming the hair bundle are not splayed, indicating that shaft connectors are not necessary to hold the stereocilia together; however, the mice show rapid postnatal deterioration in cochlear hair-bundle structure, associated with smaller than normal transducer currents with otherwise normal adaptation properties, a progressive loss of basal-coil cochlear hair cells, and deafness. These results reveal that Ptprq is required for formation of the shaft connectors of the hair bundle, the normal maturation of cochlear hair bundles, and the long-term survival of high-frequency auditory hair cells.
Asunto(s)
Cóclea/enzimología , Células Ciliadas Auditivas/enzimología , Fosfatidilinositoles/metabolismo , Monoéster Fosfórico Hidrolasas/metabolismo , Proteínas Tirosina Fosfatasas/metabolismo , Animales , Antígenos de Superficie/metabolismo , Separación Celular , Supervivencia Celular/genética , Pollos , Cilios/enzimología , Cilios/ultraestructura , Cóclea/citología , Cóclea/embriología , Sordera/genética , Regulación del Desarrollo de la Expresión Génica , Células Ciliadas Auditivas/citología , Células Ciliadas Auditivas/ultraestructura , Mecanotransducción Celular/fisiología , Ratones , Ratones Noqueados , Ratones Transgénicos , Datos de Secuencia Molecular , Técnicas de Placa-Clamp , Faloidina , Monoéster Fosfórico Hidrolasas/genética , Proteínas Tirosina Fosfatasas/genética , ARN Mensajero/metabolismo , Proteínas Tirosina Fosfatasas Clase 3 Similares a ReceptoresRESUMEN
The distribution of the cell adhesion molecule BEN in the developing chick inner ear is described. BEN is first detected in the otic placode at stage 11. As the placode begins to invaginate, BEN becomes concentrated in a ventromedial region extending from the anterior to the posterior end of the otic pit. BEN expression levels increase in this region as the pit closes to form the otocyst, and distinct boundaries become defined along the dorsal and ventral edges of the ventromedial band of BEN expression. BEN expression also becomes concentrated dorsally within the otic epithelium as the pit closes and is observed in the condensing otic ganglion. By stage 22, the ventromedial band of BEN expression splits into two distinct regions, a small caudal patch within which the posterior crista will develop, and a larger anterior patch. By stage 26, this larger anterior patch of cells expressing BEN becomes subdivided into five separate areas corresponding to the regions within which the anterior crista, the lateral crista, the utricle, the saccule, and both the basilar papilla and lagenar macula form. Hair cells only develop within these regions defined by BEN distribution. The data suggest that the ventromedial patch of BEN expression observed from stage 11 onwards defines a single sensory competent zone from which all sensory organs of the inner ear develop. BEN immunoreactivity in the inner ear declines after stage 38. In response to noise exposure, upregulation of BEN expression is mainly detected in regions of the posthatch papilla where the damage is severe and regenerating hair cells are not observed. The regenerating hair and supporting cells do not express BEN, highlighting a molecular difference between the processes of development and regeneration.
Asunto(s)
Molécula de Adhesión Celular del Leucocito Activado/metabolismo , Embrión de Pollo/citología , Células Ciliadas Auditivas/embriología , Células Ciliadas Auditivas/fisiología , Regeneración/fisiología , Molécula de Adhesión Celular del Leucocito Activado/análisis , Animales , Moléculas de Adhesión Celular Neurona-Glia/análisis , Moléculas de Adhesión Celular Neurona-Glia/metabolismo , Pollos , Conducto Coclear/química , Conducto Coclear/embriología , Conducto Coclear/fisiología , Células Ciliadas Auditivas/química , Pérdida Auditiva Provocada por Ruido/metabolismo , Pérdida Auditiva Provocada por Ruido/patología , Procesamiento de Imagen Asistido por ComputadorRESUMEN
alpha-tectorin is an extracellular matrix molecule of the inner ear. Mice homozygous for a targeted deletion in a-tectorin have tectorial membranes that are detached from the cochlear epithelium and lack all noncollagenous matrix, but the architecture of the organ of Corti is otherwise normal. The basilar membranes of wild-type and alpha-tectorin mutant mice are tuned, but the alpha-tectorin mutants are 35 dB less sensitive. Basilar membrane responses of wild-type mice exhibit a second resonance, indicating that the tectorial membrane provides an inertial mass against which outer hair cells can exert forces. Cochlear microphonics recorded in alpha-tectorin mutants differ in both phase and symmetry relative to those of wild-type mice. Thus, the tectorial membrane ensures that outer hair cells can effectively respond to basilar membrane motion and that feedback is delivered with the appropriate gain and timing required for amplification.
Asunto(s)
Cóclea/fisiología , Proteínas de la Matriz Extracelular/genética , Marcación de Gen , Glicoproteínas de Membrana/genética , Membrana Tectoria/metabolismo , Estimulación Acústica , Animales , Umbral Auditivo/fisiología , Membrana Basilar/fisiología , Cóclea/ultraestructura , Potenciales Microfónicos de la Cóclea/genética , Epitelio/patología , Exones/genética , Matriz Extracelular/genética , Matriz Extracelular/patología , Proteínas de la Matriz Extracelular/deficiencia , Proteínas de la Matriz Extracelular/metabolismo , Retroalimentación/fisiología , Proteínas Ligadas a GPI , Células Ciliadas Auditivas/citología , Células Ciliadas Auditivas Externas/citología , Células Ciliadas Auditivas Externas/fisiología , Glicoproteínas de Membrana/deficiencia , Glicoproteínas de Membrana/metabolismo , Proteínas de la Membrana/metabolismo , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Movimiento (Física) , Neuronas Aferentes/fisiología , Emisiones Otoacústicas Espontáneas/fisiología , Percepción de la Altura Tonal/fisiología , Membrana Tectoria/patologíaRESUMEN
After noise- or drug-induced hair-cell loss, the sensory epithelia of the avian inner ear can regenerate new hair cells. Few molecular markers are available for the supporting-cell precursors of the hair cells that regenerate, and little is known about the signaling mechanisms underlying this regenerative response. Hybridoma methodology was used to obtain a monoclonal antibody (mAb) that stains the apical surface of supporting cells in the sensory epithelia of the inner ear. The mAb recognizes the supporting-cell antigen (SCA), a protein that is also found on the apical surfaces of retinal Müller cells, renal tubule cells, and intestinal brush border cells. Expression screening and molecular cloning reveal that the SCA is a novel receptor-like protein tyrosine phosphatase (RPTP), sharing similarity with human density-enhanced phosphatase, an RPTP thought to have a role in the density-dependent arrest of cell growth. In response to hair-cell damage induced by noise in vivo or hair-cell loss caused by ototoxic drug treatment in vitro, some supporting cells show a dramatic decrease in SCA expression levels on their apical surface. This decrease occurs before supporting cells are known to first enter S-phase after trauma, indicating that it may be a primary rather than a secondary response to injury. These results indicate that the SCA is a signaling molecule that may influence the potential of nonsensory supporting cells to either proliferate or differentiate into hair cells.
Asunto(s)
Antígenos de Diferenciación/genética , Células Ciliadas Auditivas/química , Células Ciliadas Auditivas/enzimología , Proteínas del Tejido Nervioso/metabolismo , Proteínas Tirosina Fosfatasas/metabolismo , Animales , Antibacterianos , Anticuerpos Monoclonales , Antígenos de Diferenciación/análisis , Antígenos de Diferenciación/inmunología , Secuencia de Bases , Diferenciación Celular/fisiología , Embrión de Pollo , ADN Complementario , Detergentes , Células Epiteliales/química , Células Epiteliales/enzimología , Células Epiteliales/ultraestructura , Técnica del Anticuerpo Fluorescente , Células Ciliadas Auditivas/ultraestructura , Pérdida Auditiva Provocada por Ruido/inducido químicamente , Pérdida Auditiva Provocada por Ruido/fisiopatología , Intestinos/química , Riñón/química , Proteínas de la Membrana/análisis , Microscopía Inmunoelectrónica , Microvellosidades/química , Microvellosidades/enzimología , Datos de Secuencia Molecular , Neomicina , Ruido/efectos adversos , Octoxinol , Proteínas Tirosina Fosfatasas Clase 5 Similares a Receptores , Receptores de Superficie Celular/metabolismo , Retina/química , Homología de Secuencia de Aminoácido , Tirosina/metabolismoRESUMEN
It is generally assumed that hair-cell numbers do not increase in the vestibular epithelia of postembryonic birds after hatching. However, for the domestic chicken, it is not known when or if hair-cell numbers ever reach a steady state level during life. The numbers of hair cells in the utricular maculae of chickens from embryonic day (E) 7 to posthatch day (PH) 112 were therefore counted directly. Hair-cell numbers increase approximately 15 fold between E7 and PH2, from an average of 1,858/macula at E7 to 27,017 at PH2. Between PH2 and PH112 hair-cell numbers increase by a further 36%, to 36,650/macula. A mathematical description of the increase in hair-cell numbers observed with time predicts a half life of 29.88 days for a utricular hair cell and a steady-state turnover value of 850 hair cells/day by approximately PH60. The patterns of hair and supporting cells in the postembryonic utricular macula were also assessed. The ratios of supporting cells and hair cells, the average number of supporting cells around each hair cell, and the average number of hair cells each supporting cell contacts at PH2, PH16 and approximately 2.5 years of age are not significantly different. In contrast to the mitotically quiescent basilar papilla where all supporting cells contact at least one hair cell, 7.6% of supporting cells in the extrastriolar region of the postembryonic utricular macula do not make apical contact with a hair cell. These results indicate that hair-cell numbers in the utricular macula increase significantly after hatching, and support the concept that contact-mediated inhibition influences the proliferative potential of inner-ear supporting cells.