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Na,K-ATPase regulates a variety of transport functions in epithelial cells. In cultures of human retinal pigment epithelial (RPE) cells, inhibition of Na,K-ATPase by ouabain and K+ depletion decreased transepithelial electrical resistance (TER) and increased permeability of tight junctions to mannitol and inulin. Electrophysiological studies demonstrated that the decrease in TER was due to an increase in paracellular shunt conductance. At the light microscopy level, this increased permeability was not accompanied by changes in the localization of the tight junction proteins ZO-1, occludin, and claudin-3. At the ultrastructural level, increased tight junction permeability correlated with a decrease in tight junction membrane contact points. Decreased tight junction membrane contact points and increased tight junction permeability were reversible in K+-repletion experiments. Confocal microscopy revealed that in control cells, Na,K-ATPase was localized at both apical and basolateral plasma membranes. K+ depletion resulted in a large reduction of apical Na,K-ATPase, and after K+repletion the apical Na,K-ATPase recovered to control levels. These results suggest a functional link exists between Na,K-ATPase and tight junction function in human RPE cells.

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Anti-vascular endothelial growth factor (anti-VEGF) therapy currently plays a central role in the treatment of numerous retinal diseases, most notably exudative age-related macular degeneration (eAMD), diabetic retinopathy and retinal vein occlusions. While offering significant functional and anatomic benefits in most patients, there exists a subset of 15–40% of eyes that fail to respond or only partially respond. For these cases, various treatment options have been explored with a range of outcomes. These options include steroid injections, laser treatment (both thermal therapy for retinal vascular diseases and photodynamic therapy for eAMD), abbreviated anti-VEGF treatment intervals, switching anti-VEGF agents and topical medications. In this article, we review the effectiveness of these treatment options along with a discussion of the current research into future directions for anti-VEGF-resistant eyes.

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1. We studied the ionic mechanism of the light-peak voltage of the DC electroretinogram (DC ERG) in an in vitro preparation of chick neural retina-retinal pigment epithelium (RPE)-choroid. The light peak originates from a depolarization of the RPE basolateral (basal) membrane, associated with an increase in its conductance. Using conventional and Cl(-)-selective microelectrodes, we tested the hypothesis that the light-peak voltage is generated by an increase in Cl- conductance (gCl) of the basolateral (basal) membrane. 2. Perfusion of the RPE basal membrane with 4,4'-diisothiocyanostilbene-2,2'-disulfonate (DIDS), a known blocker of gCl in chick RPE, suppressed both the light-peak depolarization and the accompanying conductance increase of the basal membrane. 3. Using sustained transepithelial current to clamp the basal membrane potential at different levels, we estimated the reversal potential of the light peak. At membrane potentials above the equilibrium potential for Cl- (ECl = -40 +/- 10 mV mean +/- SE), light-peak polarity was reversed. Current-voltage (I-V) curves measured in the dark and at the peak of the light peak also gave a reversal potential in the same range as ECl. In addition, shifting ECl by changing intracellular Cl- (aCli) via passage of transepithelial current or perfusing the apical side of the RPE with the Cl- uptake blocker, furosemide, shifted the light-peak reversal potential in the same direction as the change in ECl. 4. The transference number for Cl-, TCl, was estimated from step decreases in basal Cl- and increased from 0.20 +/- 0.01 in the dark to 0.31 +/- 0.01 during the light peak. These results indicate an average increase of 55% in the relative conductance of the basal membrane for Cl-. 5. Light-evoked changes in aCli, measured with Cl(-)-selective microelectrodes, were too small to account for the change in basal membrane potential during the light peak. These data strongly support the hypothesis that the light peak originates from an increase in RPE basal membrane permeability to Cl-. 6. We also obtained support for the model of Joseph and Miller that the fast-oscillation trough of the DC ERG, generated by a delayed basal membrane hyperpolarization of the RPE, originates from light-evoked modulation of the Cl- transport pathway. Perfusing either the apical side of the RPE with furosemide or the basal side with DIDS suppressed the fast oscillation. The delayed basal hyperpolarization reversed polarity at membrane potentials positive to ECl.(ABSTRACT TRUNCATED AT 400 WORDS)