SLC26A7 constitutes the thiocyanate-selective anion conductance of the basolateral membrane of the retinal pigment epithelium

Abstract

Anion channels in the retinal pigment epithelium (RPE) play an essential role in the transport of Cl- between the outer retina and the choroidal blood to regulate the ionic composition and volume of the subretinal fluid that surrounds the photoreceptor outer segments. Recently, we reported that the anion conductance of the mouse RPE basolateral membrane is highly selective for the biologically-active anion thiocyanate (SCN-), a property that does not correspond with any of the Cl- channels that have been found to be expressed in the RPE to date. The purpose of this study was to determine the extent to which SLC26A7, a SCN- permeable-anion exchanger/channel that was reported to be expressed in human RPE, contributes to the RPE basolateral anion conductance. We show by qPCR that Slc26a7 is highly expressed in mouse RPE compared to other members of the Slc26 gene family and Cl- channel genes known to be expressed in the RPE. By applying immunofluorescence microscopy to mouse retinal sections and isolated cells, we localized SLC26A7 to the RPE basolateral membrane. Finally, we performed whole-cell and excised patch recordings from RPE cells acutely isolated from Slc26a7 knock out mice to show that the SCN- conductance and permeability of its basolateral membrane are dramatically smaller relative to wild type mouse RPE cells. These findings establish SLC26A7 as the SCN--selective conductance of the RPE basolateral membrane and provide new insight into the physiology of an anion channel that may participate in anion transport and pH regulation by the RPE.

Single-Cell RNA Sequencing Reveals Molecular Features of Heterogeneity in the Murine Retinal Pigment Epithelium

“…Single cells obtained by this approach were heavily pigmented and often exhibited two lobes of variable size ( Figure 1 C). This shape is consistent with an apical and basal cellular domain separated by a junctional actin band as observed in other studies, in which single RPE cells were isolated from native or cultured sheets [ 49 , 50 , 51 ]. FACS-purified cells were concentrated and applied to the wells of an scRNA-seq chip to create a bar-coded cDNA library for sequencing ( Figure 1 A).…”

Single-cell RNA sequencing reveals molecular features of postnatal maturation in the murine retinal pigment epithelium

“…Single cells obtained by this approach were heavily pigmented and often exhibited two lobes of variable size (Figure 1C). This shape is consistent with an apical and basal cellular domain separated by a junctional actin band as observed in other studies, in which single RPE cells were isolated from native or cultured sheets [37][38][39]. FACS-purified cells were concentrated and applied to the wells of an scRNA-seq chip to create a bar-coded cDNA library for sequencing (Figure 1A).…”