RESUMEN
A noise-induced loss of inner hair cell (IHC) - auditory nerve synaptic connections has been suggested as a factor that can trigger the progression of maladaptive plastic changes leading to noise-induced tinnitus. The present study used a military relevant small arms fire (SAF)-like noise (50 biphasic impulses over 2.5â¯min at 152â¯dB SPL given unilaterally to the right ear) to induce loss (â¼1/3) of IHC synaptic ribbons (associated with synapse loss) in rat cochleae with only minor (less than 10%) loss of outer hair cells. Approximately half of the noise-exposed rats showed poorer Gap Detection post-noise, a behavioral indication suggesting the presence of tinnitus. There was significantly greater loss of IHC ribbons in noise-exposed rats with reduced Gap Detection compared to noise-exposed rats retaining normal Gap Detection. We have previously shown systemic administration of piribedil, memantine, and/or ACEMg significantly reduced loss of IHC ribbons induced by a 3â¯h 4â¯kHz octave band 117â¯dB (SPL) noise. The present study examined if this treatment would also reduce ribbon loss from the SAF-like noise exposure and if this would prevent the reduced Gap Detection. As in the previous study, piribedil, memantine, and ACEMg treatment significantly reduced the noise-induced loss of ribbons, such that it was no longer significantly different from normal. However, it did not prevent development of the reduced Gap Detection indication of tinnitus in all treated noise-exposed rats, reducing the incidence but not reaching significance.
Asunto(s)
Umbral Auditivo/fisiología , Sordera/fisiopatología , Células Ciliadas Auditivas Externas/fisiología , Pérdida Auditiva Provocada por Ruido/fisiopatología , Animales , Potenciales Evocados Auditivos del Tronco Encefálico/fisiología , Masculino , Ruido , Ratas Sprague-DawleyRESUMEN
The human deafness-pigmentation syndromes, Waardenburg syndrome (WS) type 2a, and Tietz syndrome are characterized by profound deafness but only partial cutaneous pigmentary abnormalities. Both syndromes are caused by mutations in MITF. To illuminate differences between cutaneous and otic melanocytes in these syndromes, their development and survival in heterozygous Microphthalmia-White (Mitf(Mi-wh) /+) mice were studied and hearing function of these mice characterized. Mitf(Mi-wh) /+ mice have a profound hearing deficit, characterized by elevated auditory brainstem response thresholds, reduced distortion product otoacoustic emissions, absent endocochlear potential, loss of outer hair cells, and stria vascularis abnormalities. Mitf(Mi-wh) /+ embryos have fewer melanoblasts during embryonic development than their wild-type littermates. Although cochlear melanocytes are present at birth, they disappear from the Mitf(Mi-wh) /+ cochlea between P1 and P7. These findings may provide insight into the mechanism of melanocyte and hearing loss in human deafness-pigmentation syndromes such as WS and Tietz syndrome and illustrate differences between otic and follicular melanocytes.
Asunto(s)
Albinismo Oculocutáneo/fisiopatología , Sordera/fisiopatología , Audición/fisiología , Heterocigoto , Factor de Transcripción Asociado a Microftalmía/genética , Síndrome de Waardenburg/fisiopatología , Potenciales de Acción/fisiología , Albinismo Oculocutáneo/genética , Albinismo Oculocutáneo/patología , Animales , Animales Recién Nacidos , Sordera/genética , Sordera/patología , Modelos Animales de Enfermedad , Embrión de Mamíferos/metabolismo , Embrión de Mamíferos/patología , Potenciales Evocados Auditivos del Tronco Encefálico/fisiología , Células Ciliadas Auditivas Externas/metabolismo , Células Ciliadas Auditivas Externas/patología , Humanos , Melanocitos/metabolismo , Melanocitos/patología , Ratones , Ratones Endogámicos C57BL , Ratones Transgénicos , Emisiones Otoacústicas Espontáneas/fisiología , Estría Vascular/metabolismo , Estría Vascular/patología , Síndrome de Waardenburg/genética , Síndrome de Waardenburg/patologíaRESUMEN
Cyclodextrins are sugar compounds that are increasingly finding medicinal uses due to their ability to complex with hydrophobic molecules. One cyclodextrin in particular, 2-hydroxypropyl-ß-cyclodextrin (HPßCD), is used as a carrier to solubilize lipophilic drugs and is itself being considered as a therapeutic agent for treatment of Niemann-Pick Type C disease, due to its ability to mobilize cholesterol. Results from toxicological studies suggest that HPßCD is generally safe, but a recent study has found that it causes hearing loss in cats. Whether the hearing loss occurred via death of cochlear hair cells, rendering it permanent, was unexplored. In the present study, we examined peripheral auditory function and cochlear histology in mice after subcutaneous injection of HPßCD to test for hearing loss and correlate any observed auditory deficits with histological findings. On average, auditory brainstem response thresholds were elevated at 4, 16, and 32 kHz in mice one week after treatment with 8,000 mg/kg. In severely affected mice all outer hair cells were missing in the basal half of the cochlea. In many cases, surviving hair cells in the cochlear apex exhibited abnormal punctate distribution of the motor protein prestin, suggesting long term changes to membrane composition and integrity. Mice given a lower dose of 4,000 mg/kg exhibited hearing loss only after repeated doses, but these threshold shifts were temporary. Therefore, cyclodextrin-induced hearing loss was complex, involving cell death and other more subtle influences on cochlear physiology.
Asunto(s)
Anticolesterolemiantes/efectos adversos , Células Ciliadas Auditivas/efectos de los fármacos , Pérdida Auditiva/inducido químicamente , beta-Ciclodextrinas/efectos adversos , 2-Hidroxipropil-beta-Ciclodextrina , Animales , Anticolesterolemiantes/administración & dosificación , Anticolesterolemiantes/farmacología , Muerte Celular/efectos de los fármacos , Relación Dosis-Respuesta a Droga , Esquema de Medicación , Evaluación Preclínica de Medicamentos , Células Ciliadas Auditivas/fisiología , Pérdida Auditiva/patología , Infusiones Parenterales , Ratones , beta-Ciclodextrinas/administración & dosificación , beta-Ciclodextrinas/farmacologíaRESUMEN
Oxidative stress has been linked to noise- and drug-induced as well as age-related hearing loss. Antioxidants can attenuate the decline of cochlear structure and function after exposure to noise or drugs, but it is debated as to whether they can protect from age-related hearing loss. In a long-term longitudinal study, 10-month-old female CBA/J mice were placed on either a control or antioxidant-enriched diet and monitored through 24 months of age. Supplementation with vitamins A, C, and E, L-carnitine, and α-lipoic acid significantly increased the antioxidant capacity of inner ear tissues. However, by 24 months of age, the magnitude of hearing loss was equal between the two groups. Likewise, there were no significant differences in hair cell loss or degeneration of spiral ganglion cells. We conclude that dietary manipulations can alter cochlear antioxidant capacity but do not ameliorate age-related sensorineural hearing loss in the CBA/J mouse.
Asunto(s)
Antioxidantes/farmacología , Suplementos Dietéticos , Estrés Oxidativo/efectos de los fármacos , Presbiacusia/dietoterapia , Presbiacusia/prevención & control , Animales , Modelos Animales de Enfermedad , Progresión de la Enfermedad , Femenino , Estudios Longitudinales , Ratones , Ratones Endogámicos CBA , Estrés Oxidativo/fisiologíaRESUMEN
Heterozygous mutations in the gene encoding chromodomain-DNA-binding-protein 7 (CHD7) cause CHARGE syndrome, a multiple anomaly condition which includes vestibular dysfunction and hearing loss. Mice with heterozygous Chd7 mutations exhibit semicircular canal dysgenesis and abnormal inner ear neurogenesis, and are an excellent model of CHARGE syndrome. Here we characterized Chd7 expression in mature middle and inner ears, analyzed morphological features of mutant ears and tested whether Chd7 mutant mice have altered responses to noise exposure and correlated those responses to inner and middle ear structure. We found that Chd7 is highly expressed in mature inner and outer hair cells, spiral ganglion neurons, vestibular sensory epithelia and middle ear ossicles. There were no obvious defects in individual hair cell morphology by prestin immunostaining or scanning electron microscopy, and cochlear innervation appeared normal in Chd7(Gt)(/+) mice. Hearing thresholds by auditory brainstem response (ABR) testing were elevated at 4 and 16 kHz in Chd7(Gt)(/+) mice, and there were reduced distortion product otoacoustic emissions (DPOAE). Exposure of Chd7(Gt)(/+) mice to broadband noise resulted in variable degrees of hair cell loss which inversely correlated with severity of stapedial defects. The degrees of hair cell loss and threshold shifts after noise exposure were more severe in wild type mice than in mutants. Together, these data indicate that Chd7(Gt)(/+) mice have combined conductive and sensorineural hearing loss, correlating with changes in both middle and inner ears.
Asunto(s)
Síndrome CHARGE/enzimología , Proteínas de Unión al ADN/metabolismo , Oído Interno/enzimología , Oído Medio/enzimología , Pérdida Auditiva Conductiva/enzimología , Pérdida Auditiva Sensorineural/enzimología , Estimulación Acústica , Factores de Edad , Animales , Umbral Auditivo , Síndrome CHARGE/genética , Síndrome CHARGE/patología , Síndrome CHARGE/fisiopatología , Proteínas de Unión al ADN/genética , Modelos Animales de Enfermedad , Oído Interno/anomalías , Oído Interno/fisiopatología , Oído Interno/ultraestructura , Oído Medio/anomalías , Oído Medio/fisiopatología , Oído Medio/ultraestructura , Potenciales Evocados Auditivos del Tronco Encefálico , Femenino , Genes Reporteros , Pérdida Auditiva Conductiva/genética , Pérdida Auditiva Conductiva/patología , Pérdida Auditiva Conductiva/fisiopatología , Pérdida Auditiva Sensorineural/genética , Pérdida Auditiva Sensorineural/patología , Pérdida Auditiva Sensorineural/fisiopatología , Inmunohistoquímica , Masculino , Ratones , Ratones de la Cepa 129 , Ratones Transgénicos , Microscopía Electrónica de Rastreo , Proteínas Motoras Moleculares/metabolismo , Mutación , Ruido , Emisiones Otoacústicas Espontáneas , Regiones Promotoras Genéticas , beta-Galactosidasa/genética , beta-Galactosidasa/metabolismoRESUMEN
Alström Syndrome is a life-threatening disease characterized primarily by numerous metabolic abnormalities, retinal degeneration, cardiomyopathy, kidney and liver disease, and sensorineural hearing loss. The cellular localization of the affected protein, ALMS1, has suggested roles in ciliary function and/or ciliogenesis. We have investigated the role of ALMS1 in the cochlea and the pathogenesis of hearing loss in Alström Syndrome. In neonatal rat organ of Corti, ALMS1 was localized to the basal bodies of hair cells and supporting cells. ALMS1 was also evident at the basal bodies of differentiating fibrocytes and marginal cells in the lateral wall. Centriolar ALMS1 expression was retained into maturity. In Alms1-disrupted mice, which recapitulate the neurosensory deficits of human Alström Syndrome, cochleae displayed several cyto-architectural defects including abnormalities in the shape and orientation of hair cell stereociliary bundles. Developing hair cells were ciliated, suggesting that ciliogenesis was largely normal. In adult mice, in addition to bundle abnormalities, there was an accelerated loss of outer hair cells and the progressive appearance of large lesions in stria vascularis. Although the mice progressively lost distortion product otoacoustic emissions, suggesting defects in outer hair cell amplification, their endocochlear potentials were normal, indicating the strial atrophy did not affect its function. These results identify previously unrecognized cochlear histopathologies associated with this ciliopathy that (i) implicate ALMS1 in planar cell polarity signaling and (ii) suggest that the loss of outer hair cells causes the majority of the hearing loss in Alström Syndrome.
Asunto(s)
Síndrome de Alstrom/metabolismo , Síndrome de Alstrom/patología , Cóclea/ultraestructura , Proteínas de Unión al ADN/metabolismo , Células Ciliadas Auditivas/metabolismo , Células Ciliadas Auditivas/ultraestructura , Pérdida Auditiva/genética , Pérdida Auditiva/patología , Síndrome de Alstrom/genética , Animales , Proteínas de Ciclo Celular , Diferenciación Celular , Polaridad Celular , Centriolos , Cilios/ultraestructura , Proteínas de Unión al ADN/genética , Técnica del Anticuerpo Fluorescente , Pérdida Auditiva/metabolismo , Ratones , Ratones Noqueados , Microscopía Electrónica , Órgano Espiral/ultraestructura , Ratas , Ratas Sprague-Dawley , Transducción de Señal , Estría Vascular/ultraestructuraRESUMEN
Commercially obtained aged male CBA/J mice presented a complex pattern of hearing loss and morphological changes. A significant threshold shift in auditory brainstem responses (ABR) occurred at 3 months of age at 4 kHz without apparent loss of hair cells, rising slowly at later ages accompanied by loss of apical hair cells. A delayed high-frequency deficit started at 24 kHz around the age of 12 months. At 20-26 months, threshold shifts at 12 and 24 kHz and the accompanying hair cell loss at the base of the cochlea were highly variable with some animals appearing almost normal and others showing large deficits. Spiral ganglion cells degenerated by 18 months in all regions of the cochlea, with cell density reduced by approximately 25%. There was no degeneration of the stria vascularis and the endocochlear potential remained stable from 3 to 25 months of age regardless of whether the animals had normal or highly elevated ABR thresholds. The slow high-frequency hearing loss combined with a modest reduction of ganglion cell density and an unchanged endocochlear potential suggest sensorineural presbycusis. The superimposed early hearing loss at low frequencies, which is not seen in animals bred in-house, may complicate the use of these animals as a presbycusis model.
Asunto(s)
Envejecimiento/patología , Presbiacusia/patología , Animales , Umbral Auditivo , Cóclea/patología , Potenciales Microfónicos de la Cóclea , Modelos Animales de Enfermedad , Potenciales Evocados Auditivos del Tronco Encefálico , Células Ciliadas Auditivas/patología , Pérdida Auditiva de Alta Frecuencia/etiología , Pérdida Auditiva de Alta Frecuencia/patología , Pérdida Auditiva de Alta Frecuencia/fisiopatología , Masculino , Ratones , Ratones Endogámicos CBA , Presbiacusia/etiología , Presbiacusia/fisiopatología , Ganglio Espiral de la Cóclea/patología , Estría Vascular/patologíaRESUMEN
The basic helix-loop-helix (bHLH) transcription factor Math5 (Atoh7) is required for retinal ganglion cell (RGC) and optic nerve development. Using Math5-lacZ knockout mice, we have identified an additional expression domain for Math5 outside the eye, in functionally connected structures of the central auditory system. In the adult hindbrain, the cytoplasmic Math5-lacZ reporter is expressed within the ventral cochlear nucleus (VCN), in a subpopulation of neurons that project to medial nucleus of the trapezoid body (MNTB), lateral superior olive (LSO), and lateral lemniscus (LL). These cells were identified as globular and small spherical bushy cells based on their morphology, abundance, distribution within the cochlear nucleus (CN), co-expression of Kv1.1, Kv3.1b and Kcnq4 potassium channels, and projection patterns within the auditory brainstem. Math5-lacZ is also expressed by cochlear root neurons in the auditory nerve. During embryonic development, Math5-lacZ was detected in precursor cells emerging from the caudal rhombic lip from embryonic day (E)12 onwards, consistent with the time course of CN neurogenesis. These cells co-express MafB and are post-mitotic. Math5 expression in the CN was verified by mRNA in situ hybridization, and the identity of positive neurons was confirmed morphologically using a Math5-Cre BAC transgene with an alkaline phosphatase reporter. The hindbrains of Math5 mutants appear grossly normal, with the exception of the CN. Although overall CN dimensions are unchanged, the lacZ-positive cells are significantly smaller in Math5 -/- mice compared to Math5 +/- mice, suggesting these neurons may function abnormally. The auditory brainstem response (ABR) of Math5 mutants was evaluated in a BALB/cJ congenic background. ABR thresholds of Math5 -/- mice were similar to those of wild-type and heterozygous mice, but the interpeak latencies for Peaks II-IV were significantly altered. These temporal changes are consistent with a higher-level auditory processing disorder involving the CN, potentially affecting the integration of binaural sensory information.
Asunto(s)
Vías Auditivas/metabolismo , Factores de Transcripción con Motivo Hélice-Asa-Hélice Básico/fisiología , Proteínas del Tejido Nervioso/fisiología , Animales , Vías Auditivas/citología , Factores de Transcripción con Motivo Hélice-Asa-Hélice Básico/genética , Bromodesoxiuridina/metabolismo , Núcleo Coclear/anatomía & histología , Potenciales Evocados Auditivos del Tronco Encefálico/genética , Ratones , Ratones Endogámicos BALB C , Ratones Transgénicos , Proteínas del Tejido Nervioso/genética , Neuronas/metabolismo , ARN Mensajero/metabolismo , Estilbamidinas/metabolismo , beta-Galactosidasa/metabolismoRESUMEN
Thyroid hormone (TH) insufficiency causes variable hearing impairment and mental deficiency in humans. Rodents lacking TH have congenital hearing deficiency that has been attributed to physiologic, morphologic, and developmental abnormalities of the auditory system. We examined four genetically defined strains of hypothyroid mice for development of hearing and response to TH replacement initiated during late gestation and continued through six weeks of age. Auditory brain stem response studies showed variable hearing impairment in homozygous mutants of each strain at three weeks of age relative to normal littermates. Mutants from three of the strains still had hearing deficiencies at six weeks of age. TH-enriched diet significantly improved hearing in three-week-old mutants of each strain relative to untreated mutants. Differences in the level of hearing impairment between the Prop1df and Pit1dw mutants, which have defects in the same developmental pathway, were determined to be due to genetic background modifier genes. Further physiologic and morphologic studies in the Cgatm1Sac strain indicated that poor hearing was due to cochlear defects. We conclude that TH supplement administered during the critical period of hearing development in mice can prevent deafness associated with congenital hypothyroidism of heterogeneous genetic etiology.
Asunto(s)
Dieta , Trastornos de la Audición/tratamiento farmacológico , Hipotiroidismo/tratamiento farmacológico , Triyodotironina/administración & dosificación , Triyodotironina/uso terapéutico , Administración Oral , Animales , Trastornos de la Audición/sangre , Trastornos de la Audición/etiología , Proteínas de Homeodominio/genética , Terapia de Reemplazo de Hormonas , Hipotiroidismo/sangre , Hipotiroidismo/complicaciones , Hipotiroidismo/genética , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos C57BL , Ratones Mutantes , Ratones Transgénicos , Modelos Biológicos , Órgano Espiral/anatomía & histología , Órgano Espiral/crecimiento & desarrollo , Emisiones Otoacústicas Espontáneas/genética , Receptores de Tirotropina/genética , Tiroxina/sangre , Factor de Transcripción Pit-1/genéticaRESUMEN
The aims of this study were to explore the correlation between DPOAE adaptation magnitude in three different guinea pig strains to examine if the genetic component affects the DPOAE adaptation magnitude. It was also to investigate the correlation between strains with certain characteristics i.e. reduced susceptibility to noise, and early onset of age-dependent hearing loss and the DPOAE adaptation magnitude. The animals were anaesthetized and the 2f1-f2 DPOAE (f1=8k Hz, and f2/f1=1.2) adaptation was established with a minimum of 144 combinations of f1; f2 where f1 was held fixed and f2 was varied in 1 dB or 0.4 dB steps. The DPOAE adaptation magnitude was defined as the difference between maximum positive level and the maximum negative level. ABRs were conducted at different age-groups (at 4, 6.3, and 12.5k Hz) to evaluate the progress of hearing thresholds by age. There was a significant difference between strains regarding the hearing loss at one year of age. There was no significant difference in DPOAE adaptation magnitude between strains included in this study and from this we conclude that the DPOAE adaptation magnitude is not a predictor for the susceptibility to noise trauma, or early onset of age-dependent hearing loss, using the methods described in this paper.
Asunto(s)
Cobayas/fisiología , Emisiones Otoacústicas Espontáneas/fisiología , Adaptación Fisiológica , Envejecimiento/fisiología , Animales , Umbral Auditivo , Modelos Animales de Enfermedad , Vías Eferentes/fisiología , Potenciales Evocados Auditivos del Tronco Encefálico , Cobayas/genética , Pérdida Auditiva Provocada por Ruido/etiología , Pérdida Auditiva Provocada por Ruido/genética , Pérdida Auditiva Provocada por Ruido/fisiopatología , Emisiones Otoacústicas Espontáneas/genética , Presbiacusia/etiología , Presbiacusia/genética , Presbiacusia/fisiopatología , Reflejo Acústico , Especificidad de la EspecieRESUMEN
We applied the dopaminergic (DA) neurotoxin 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) to the guinea pig cochlear perilymph. Immunolabeling of lateral olivocochlear (LOC) neurons using antibodies against synaptophysin was reduced after the MPTP treatment. In contrast, labeling of the medial olivocochlear innervation remained intact. As after brainstem lesions of the lateral superior olive (LSO), the site of origin of the LOC neurons, the main effect of disrupting LOC innervation of the cochlea via MPTP was a depression of the amplitude of the compound action potential (CAP). CAP amplitude depression was similar to that produced by LSO lesions. Latency of the N1 component of the CAP, and distortion product otoacoustic emission amplitude and adaptation were unchanged by the MPTP treatment. This technique for selectively lesioning descending LOC efferents provides a new opportunity for examining LOC modulation of afferent activity and behavioral measures of perception.
Asunto(s)
Nervio Coclear/fisiología , Núcleo Coclear/fisiología , Dopamina/fisiología , Potenciales Evocados Auditivos del Tronco Encefálico/fisiología , Núcleo Olivar/fisiología , 1-Metil-4-fenil-1,2,3,6-Tetrahidropiridina/farmacología , Potenciales de Acción , Adaptación Fisiológica/efectos de los fármacos , Adaptación Fisiológica/fisiología , Animales , Núcleo Coclear/patología , Desnervación , Dopaminérgicos/farmacología , Potenciales Evocados Auditivos del Tronco Encefálico/efectos de los fármacos , Femenino , Cobayas , Inmunohistoquímica , Masculino , Neurotoxinas/farmacología , Núcleo Olivar/patología , Emisiones Otoacústicas Espontáneas/efectos de los fármacos , Emisiones Otoacústicas Espontáneas/fisiologíaRESUMEN
Rapid efferent adaptation of the distortion product otoacoustic emission (DPOAE) predicts susceptibility to noise-induced damage, and is linked to the concentration of the efferent receptor (alpha9). Maximum adaptation occurs at intense primary levels, rapidly switching from positive to negative orientation in a very narrow (2 dB) range of F1 and F2 levels. Aminoglycosides are commonly used antibiotics, with the undesirable side-effect of ototoxicity. Susceptibility to hair cell damage from the aminoglycoside gentamicin can be quite variable, even within a single strain and species of animal. Since one of gentamicin's first sites of action in the outer hair cell (OHC) is at the efferent receptor, it is possible that efferent activity could be a predictor of susceptibility to gentamicin induced damage. Significant sex-related differences were found in two strains of guinea pigs when treated with gentamicin. Female guinea pigs were more susceptible both to systemic effects and to specific ototoxic effects. Efferent-mediated DPOAE adaptation served as a predictor of sensitivity to aminoglycoside damage, predicting both number of days before onset of deafness in male animals, and predicting final threshold shifts from gentamicin doses which produced variable results.
Asunto(s)
Adaptación Fisiológica , Aminoglicósidos/toxicidad , Antibacterianos/toxicidad , Pérdida Auditiva/inducido químicamente , Neuronas Eferentes/efectos de los fármacos , Emisiones Otoacústicas Espontáneas , Adaptación Fisiológica/efectos de los fármacos , Animales , Umbral Auditivo/efectos de los fármacos , Relación Dosis-Respuesta a Droga , Potenciales Evocados Auditivos del Tronco Encefálico , Femenino , Gentamicinas/toxicidad , Cobayas , Masculino , Emisiones Otoacústicas Espontáneas/efectos de los fármacos , Emisiones Otoacústicas Espontáneas/fisiología , Análisis de Regresión , Factores Sexuales , Organismos Libres de Patógenos EspecíficosRESUMEN
Tight junctions (TJs) create ion-selective paracellular permeability barriers between extracellular compartments. In the organ of Corti of the inner ear, TJs of the reticular lamina separate K(+)-rich endolymph and Na(+)-rich perilymph. In humans, mutations of the gene encoding claudin 14 TJ protein cause profound deafness but the underlying pathogenesis is unknown. To explore the role of claudin 14 in the inner ear and in other tissues we created a mouse model by a targeted deletion of Cldn14. In the targeted allele a lacZ cassette is expressed under the Cldn14 promoter. In Cldn14-lacZ heterozygous mice beta-galactosidase activity was detected in cochlear inner and outer hair cells and supporting cells, in the collecting ducts of the kidney, and around the lobules of the liver. Cldn14-null mice have a normal endocochlear potential but are deaf due to rapid degeneration of cochlear outer hair cells, followed by slower degeneration of the inner hair cells, during the first 3 weeks of life. Monolayers of MDCK cells expressing claudin 14 show a 6-fold increase in the transepithelial electrical resistance by decreasing paracellular permeability for cations. In wild type mice, claudin 14 was immunolocalized at hair cell and supporting cell TJs. Our data suggest that the TJ complex at the apex of the reticular lamina requires claudin 14 as a cation-restrictive barrier to maintain the proper ionic composition of the fluid surrounding the basolateral surface of outer hair cells.