Expression of anion exchangers in cultured human endolymphatic sac epithelia

Abstract

The endolymphatic sac (ES) is part of the membranous labyrinth in the inner ear that plays an important role in maintaining homeostasis of the endolymphatic fluid system. However, the exact mechanism of fluid volume and pH regulation is not fully understood yet. We aimed to demonstrate the expression of various anion exchangers (AE), including pendrin, in cultured human endolymphatic sac epithelial (HESE) cells. Endolymphatic sac specimens were harvested during acoustic neuroma surgery (n=24) using the translabyrinthine approach and then subcultured with high-epidermal growth factor (EGF) (25 ng/mL) media and differentiated using low-EGF (0.5 ng/mL) media. The cultured cells were classified according to the morphology on TEM. The Cl-/HCO3- exchanger activity was assessed by pHi measurement using pH sensitive dye 2΄, 7΄-bis (2-carboxyethyl)-5(6) –carboxyfluorescein (BCECF/AM). We performed RT-PCR and immunohistochemical staining for AEs. We determined that 7.3 ± 6.7% of cells differentiated into mitochodria-rich cells and 50.2 ± 15.1 of cells differentiated into ribosome-rich cells. bAE3, AE4, SLC26A4, SLC26A6, and SLC26A11 were also expressed in cultured HESE cells. The cultured cells had Cl-/HCO3- and Cl-/formate exchange activity on the luminal membrane, which is sensitive to anion channel inhibitors (DIDS 500 μM). Furthermore, we showed that pendrin (SLC26A4) was expressed in cultured HESE cell membranes. Our results suggest that AEs, including pendrin, are expressed in epithelia of ES and may have role in maintaining ionic homeostasis, and the HESE culture system are useful for uncovering the functional role of ES epithelial cells.