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Objective@#To test the mechanism and upstream pathway of outer hair cell apoptosis in Cadherin 23 (Cdh23) gene mutant mice.@*Method@#The mutant Cdh23erl/erl(erl) mice were collected as the study group, while the C57BL/6J (B6) mice were chosen as the control group. A total of 70 mice per group were used in this study. The study group and control group underwent auditory-evoked brainstem response (ABR) tests at the same age. The terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay was performed to detect outer hair cell(OHC) apoptosis. The qRT-PCR was conducted to test the expression of ER stress markers immunoglobulin-binding protein (BiP) and C/EBP homologous protein (CHOP) mRNA. The expression and location of BiP and CHOP protein in OHC were detected by immunostaining. The expression of BiP protein in cochleae was identified by Western blot. The expression and location of CDH23 protein in OHC were discovered by immunostaining.@*Results@#The ABR thresholds in erl mice were significantly higher than those in B6 mice at the age of 1 and 3 months (both P<0.05). The surface preparation with TUNEL staining confirmed OHC apoptosis in erl mouse cochleae which showed a higher TUNEL positive cell ratio than B6 mouse(t=11.291, P<0.01). The ER stress marker Bip and Chop mRNA were upregulated in the erl mouse inner ear, when compared with those in the B6 mouse(both P<0.05). The BiP protein extracted from the erl mouse cochleae was significantly higher than that of B6 mouse measured by Western blot (t=3.66, P=0.02). Immunostaining showed that BiP and CHOP were highly detected in the OHC in erl mouse cochleae, and was mainly detected in the perinuclear region of OHC. However, a bare BiP and CHOP signal were shown in B6 mouse cochleae. The CDH23 protein was specifically localized at the top of the OHC in B6 mice, indicating the localization of the tip links in hair bundle stereocilia. On the contrary, the CDH23erl protein was found to be localized from the top to the nuclei of the OHC in erl mice. Portions of the CDH23erl proteins failed to reach the top of the hair bundles and remained in the OHC cytoplasm.@*Conclusion@#As the downstream response of the Cdh23 gene mutation, portions of the mutant CDH23erl protein was accumulated in ER lumen resulting in the increase of ER loading and ultimately triggered ER stress and hair cell apoptosis in erl mouse cochleae.
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Objective@#To explore the regulation and mechanism of Prestin protein by identifying the proteins interacted with Prestin in cochlear outer hair cell(OHC) and analyzing their biological function.@*Methods@#Co-immunoprecipitation combined mass spectrometry technology was used to isolate and identify the proteins interacted with Prestin protein of OHC, bioinformatics was used to construct Prestin protein interaction network. The proteins interacted with Prestin in OHC of guinea pig were determined by matching primary interaction mass spectrometry with protein interaction network, and annotated their functions.@*Results@#The results of co-immunoprecipitation combined with mass spectrometry showed that 116 kinds of credible proteins could interact with Prestin. By constructing Prestin protein interaction network, matching the results of mass spectrometry and analyzing of sub-cellular localization, eight kinds of proteins were confirmed that they interacted with Prestin directly, namely EEF2, HSP90AB1, FN1, FLNA, EEF1A1, HSP90B1, ATP5A1, and ERH, respectively, which were mainly involved in the synthesis and transportation, transmembrane folding and localization, structural stability and signal transduction of Prestin protein.@*Conclusion@#EEF2, HSP90AB1, FN1, FLNA, EEF1A1, HSP90B1, ATP5A1 and ERH provide molecular basis for sensory amplification function of OHCs by participating in biotransformation, transmembrane folding and localization, signal transduction and other biological processes of Prestin protein.
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RESUMO Objetivo: investigar o efeito supressor das emissões otoacústicas por estímulos transientes em indivíduos com queixa zumbido e audiometria normal e analisar sua relação com as variáveis idade, sexo, lateralidade do zumbido e grau de incômodo. Métodos: foram avaliados 60 sujeitos, 14 do gênero masculino e 46 do gênero feminino, entre 20 e 59 anos de idade, sendo 30 com queixa de zumbido (grupo experimental) e 30 sem zumbido (grupo controle). Foi realizada a pesquisa da supressão das emissões otoacústicas por estímulos transientes, para ruído branco de 50 dBNA, na condição contralateral nas bandas de frequência de 700, 1000, 1400, 2000, 2800 and 4000Hz. Resultado: no grupo experimental, a supressão das emissões otoacústicas transientes média variou de 2,14 a 4,38. No grupo controle o valor médio da supressão das emissões otoacústicas transientes variou de 2,27 a 4,88. Conclusão: os valores de supressão das emissões otoacústicas foram semelhantes nos indivíduos com e sem zumbido, embora o grupo com o sintoma tenha tido resultados menores, sugerindo pior desempenho do Complexo Olivar Superior.
ABSTRACT Purpose: to investigate the suppressive effect of transient-evoked otoacoustic emissions in subjects with tinnitus complaint and normal audiometry and to analyze the relation to age, gender, laterality of tinnitus and its degree of discomfort. Methods: we assessed 60 subjects, 14 males and 46 females, aged between 20 and 59 years, 30 with tinnitus (experimental group) and 30 without tinnitus complaint (control group). The suppression of transient-evoked otoacoustic emissions was investigated with contralateral white noise at 50 dBHL at the frequency bands of 700, 1000, 1400, 2000, 2800 and 4000Hz. Results: the mean value for the suppression of transient-evoked otoacoustic emissions in the experimental group ranged from 2.14 to 4.38. In the control group, the mean value for suppression of transient-evoked otoacoustic emissions ranged from 2.27 to 4.88. Conclusion: suppression values of otoacoustic emissions were similar in subjects with and without tinnitus, although the results of the tinnitus group were lower, suggesting worse performance of the Superior Olivary Complex.
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Objetivo verificar a ocorrência de perda auditiva sensorioneural em crianças com baixo nível de exposição cumulativa ao chumbo. Métodos 156 crianças intoxicadas por chumbo, 94 do sexo masculino e 62 do sexo feminino, na faixa etária entre 18 meses a 14 anos e 5 meses, foram submetidas a análise longitudinal do nível de Plumbemia em sangue, bem como audiometria tonal liminar e emissões otoacústicas evocadas por estímulo transiente. Resultados a população pesquisada apresentou um valor médio de Plumbemia estimada de 12,2±5,7mg/dL (faixa entre 2,4-33mg/dL); todas as crianças apresentaram resposta normal na audiometria tonal liminar em 20 dBNA nas frequências testadas, 0,5; 1; 2 e 4 kHz, para ambas as orelhas; as emissões otoacústicas evocadas por estímulo transiente estiveram presentes para todas as frequências bilateralmente, nas 79 crianças pesquisadas. Conclusão não foi constatada perda auditiva sensorioneural em crianças com histórico de baixo nível de exposição cumulativa por chumbo, assim como não foi encontrada lesão de células ciliadas externas na cóclea, mesmo que subclínicas. .
Purpose to verify the occurrence of sensorineural hearing loss in children with low level of cumulative lead exposure. Methods 156 lead-poisoned children, 94 males and 62 females, ranging in age from 18 months old to 14 years and 5 months old were subjected to analysis of longitudinal lead level in blood as well as pure tone audiometry and transient evoked otoacoustic emissions. Results the population studied had a mean estimated blood lead level of 12,2±5,7mg/dL (range between 2,4 and 33mg/dL); all children had a normal response in pure tone audiometry at 20 dBHL in the frequencies tested, 0.5, 1, 2 and 4 kHz, in both ears; the transient evoked otoacoustic emissions were presented for all frequencies bilaterally in 79 children surveyed. Conclusion there has been no hearing loss in children with a history of low cumulative lead exposure, as there was no injury of cochlear outer hair cells, even if subclinical. .