Reduced postnatal expression of cochlear Connexin26 induces hearing loss and affects the developmental status of pillar cells in a dose-dependent manner.

Mutations in the GJB2 gene (which encodes Connexin26 (Cx26)) are the most common cause of non-syndromic deafness. Previous studies showed that an extensive knockout of the Gjb2 gene in cochlear epithelium can cause severe deafness, significant hair cell (HC) loss and failure of pillar cells (a type of supporting cell, PCs) to differentiate in mice. This study aimed to establish different mouse models with gradient reductions of cochlear Cx26 expression and to investigate the effect of different reduced levels of cochlear Cx26 expression on hearing and development of PCs. According to the reduction in the levels of cochlear Cx26, these models were named high knockdown (KD), middle KD and low KD group. In the low KD group, the mice showed normal hearing and well-developed PCs. In the high KD group, up to 90 percent of supporting cells (SCs) lost Cx26 expression. These mice exhibited severe deafness, rapid hair cell degeneration and juvenile PCs. In the middle KD group, nearly half of SCs lost Cx26 expression. However, these mice showed a moderate deafness and a late-onset hair cell loss. Moreover, nearly all the PCs in mice of this group were in a partially differentiated state. These results indicated that reduction of postnatal expression of cochlear Cx26 induces hearing loss in a dose-dependent manner. Null Cx26 in a few SCs affects the developmental status of PCs and the hair cell degeneration pattern. The abnormal developmental status of PCs may be a potential cause of Gjb2-related hearing loss.

The spatial distribution pattern of Connexin26 expression in supporting cells and its role in outer hair cell survival.

Mutations in the GJB2 gene (which encodes Connexin26 (Cx26)) account for about a quarter of all cases of non-syndromic deafness. Previous studies have indicated that knockout (KO) of Gjb2 gene during early postnatal days can cause outer hair cell (OHC) loss in mouse models. However, the postnatal spatial distribution pattern of Cx26 in different types of supporting cells (SCs) and the role of such distributions for the survival of OHCs is still obscure. In this study, the spatial distribution patterns of Cx26 in SCs were observed, and based on these observations different spatial Cx26-null mouse models were established in order to determine the effect of changes in the spatial distribution of Cx26 in SCs on the survival of OHCs. At postnatal day (P)3, unlike the synchronous expression of Cx26 along both longitudinal and radial boundaries of most types of SCs, Cx26 expression was primarily observed along the longitudinal boundaries of rows of Deiter's cells (DCs). From P5 to P7, radial expression of Cx26 was gradually observed between adjacent rows of DCs. When Gjb2 gene was knocked out at random in different types of SCs, about 40% of the total DCs lost Cx26 expression and these Cx26-null DCs were distributed randomly in all three rows of DCs. The mice in this randomly Cx26-null group showed normal hearing and no significant OHC loss. When using a longitudinal KO pattern to induce knockout of Gjb2 gene specifically in the third row of DCs, about 33% of the total DCs lost Cx26 expression in this specific longitudinally Cx26-null group. The mice in this group showed late-onset hearing loss and significant OHC loss, however, the morphology of corresponding DCs was slightly altered. In both experimental groups, no substantial DC loss was observed. These results indicate that longitudinal Cx26-based channels are predominant in DCs during P3-P5. The Cx26 expression along rows of DCs might play a key role in the survival of OHCs, but this longitudinal KO pattern in DCs has a limited effect on DC survival or on its postnatal development.