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Loss-of function mutations in transmembrane inner ear expressed (Tmie/TMIE) gene have been shown to cause deafness in mice and humans (DFNB6). However, the functional roles of TMIE in the cochlea remain unclear. A primary step toward the understanding of the role of TMIE in hearing and its dysfunction is the documentation of its cellular and sub-cellular location within the cochlea, the auditory organ. In this study, we located and determined the cellular expression of Tmie within the rat cochlea using a polyclonal anti-Tmie antibody. The anti-Tmie antibody identified a specific band of 17 kDa in a variety of rat tissues by using Western blot analyses. The expression products of Tmie were also detected in the spiral limbus, spiral ligament, organ of Corti, and stria vascularis by immunohistochemistry analysis and RT-PCR. Our results point out the presence and localization of Tmie products in the cochlea of rat. Knowledge of spatial distribution of Tmie will provide important insight into the mechanisms that lead to deafness due to mutations in the TMIE gene.

Multiple types of connexin (Cxs) products, including Cx26, Cx30, Cx31, and Cx43, are found by immunolabeling in the mature cochlea. The transcript of Cx29, a newly discovered member of Cx gene family, was also discovered in the cochlea by cDNA macroarray hybridization. However, the functional roles of Cx29 in the cochlea remain unclear. To elucidate whether the Cx29 gap junction protein epsilon 1, GJE1, is localized in the adult mouse and rat cochlea, we performed an immunohistochemistry (IHC) and reverse transcription-polymerase chain reaction (RT-PCR) analysis. GJE1 was detected in the cochlea neurons, spiral limbus, spiral ligament, organ of Corti, and stria vascularis using IHC analysis. We also show that Cx29 mRNA is present in spiral limbus, spiral ligament, organ of Corti, stria vascularis, and lateral wall by the method of RT-PCR. Higher levels of Cx29 mRNA were found in spiral ligament and spiral limbus, whereas lower level in lateral wall. Our data first provide a comprehensive and detailed pattern of Cx29 gene expression in the mouse and rat cochlea. Knowledge of spatial distribution of Cx29 also allows the identification of candidate genes for deafness and provides important insight into mechanisms that lead to deafness due to mutations in Cx29 gene.

Recessive mutations of PDS gene are the common causes of Pendred syndrome and non-syndromic hearing loss associated with temporal bone abnormalities ranging from isolated enlargement of the vestibular aqueduct (EVA) to Mondini dysplasia. In this study we evaluate the relationship between EVA and Mondini dysplasia in 10 prelingual deaf patients and PDS gene mutation. One of three mutations, IVS7-2A-->G, IVS16-6G-->A or IVS15+5G-->A, was identified in the PDS gene in each patient. In family studies of four probands with the IVS7-2A-->G mutation, we found that this mutation was inherited from the same mutant alleles of parental origin. The effect of IVS7-2A-->G mutation on PDS gene expression was determined by reverse transcription and polymerase chain reaction (RT-PCR). Sequencing of the RT-PCR products revealed that the PDS transcripts from the allele with IVS7-2A-->G mutation lose the entire exon 8, resulting in a joining of exons 7 and 9. Deletion of the exon 8 results in frameshift and premature termination of translation. Haplotype analysis showed a significant haplotype shared among the family members carrying IVS7-2A-->G mutation, suggesting that they may be derived from a common ancestor. Our results provide evidence that hearing loss with EVA and Mondini dysplasia may be caused by splice-site mutation in the PDS gene.