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Mechanisms of Ion Transport across the Mouse Retinal Pigment Epithelium Measured in Vitro

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dc.contributor Landspitali - The National University Hospital of Iceland
dc.contributor.author Skarphedinsdottir, Sunna Bjorg
dc.contributor.author Eysteinsson, Þór
dc.contributor.author Árnason, Sighvatur Sævar
dc.date.accessioned 2022-08-04T01:02:23Z
dc.date.available 2022-08-04T01:02:23Z
dc.date.issued 2020-06
dc.identifier.citation Skarphedinsdottir , S B , Eysteinsson , Þ & Árnason , S S 2020 , ' Mechanisms of Ion Transport across the Mouse Retinal Pigment Epithelium Measured in Vitro ' , Investigative Ophthalmology and Visual Science , vol. 61 , no. 6 , 31 . https://doi.org/10.1167/IOVS.61.6.31
dc.identifier.issn 0146-0404
dc.identifier.other PURE: 37504086
dc.identifier.other PURE UUID: 0242f0da-f364-4f73-afb8-fef20a9a631c
dc.identifier.other Scopus: 85086623966
dc.identifier.uri https://hdl.handle.net/20.500.11815/3306
dc.description Funding Information: Supported by grants from The Icelandic Research Council, University of Iceland Research Fund, and The Helga Jónsdot-tir and Sigurlidi Kristjánsson Memorial Fund. Publisher Copyright: © 2020 Association for Research in Vision and Ophthalmology Inc.. All rights reserved.
dc.description.abstract PURPOSE. To examine ion transport across the mouse retinal pigment epithelium (RPE), measured by the short-circuit current (ISC) and transepithelial resistance (TER). METHODS. Sheets of RPE from mice (C57BL6/J) with retina, choroid, and sclera attached were mounted in Ussing chambers (0.031-cm2 aperture) and Krebs solution. The ISC and TER were recorded with voltage clamps. Receptors implicated in ion transport were blocked or stimulated by ligands applied to both sides. RESULTS. The mean initial ISC was −12.0 ± 3.9 μA/cm2 (basolateral negative), and mean TER was 67.1 ± 8.0 ohm·cm2. RPE preparations remained stable for 3 hours, with ISC decreasing by 0.078 ± 0,033 μA/cm2/hr. Adenosine triphosphate (100 μM) increased ISC by 2.22 ± 0.41 μA/cm2 (P = 0.003). Epinephrine (100 μM) increased ISC by 1.14 ± 0.19 μA/cm2 (P = 0.011). Bumetanide (100 μM) reduced ISC by 1.72 ± 0.73 μA/cm2 (P = 0.027). Ouabain (1 mM) induced a biphasic response: an ISC increase from −7.9 ± 2.4 to −15.49 ± 2.12 μA/cm2 and then a decrease to −3.7 ± 2.2 μA/cm2. Ouabain increased TER by 15.3 ± 4.8 ohm·cm2. These compounds were added sequentially. Apical [K+]o at zero mM transiently increased ISC by 3.36 ± 1.06 μA/cm2. Ba++ decreased ISC from −10.4 ± 3.1 to −6.6 ± 1.8 μA/cm2 (P = 0.01). Ba++ reversed the K+-free response, with Isc decreasing further from −5.65 ± 1.24 to −3.37 ± 0.79 μA/cm2 (P = 0.029). CONCLUSIONS. The ISC and TER can be recorded from the mouse RPE for 3 hours. Adrenergic and purinergic receptors affect murine RPE ion transport. Sodium–potassium adenosine triphosphatase plays a role in net ion transport across mouse RPE, and Na-K-2Cl cotransporter activity partly accounts for transepithelial ion transport. Mimicking light-induced changes, low subretinal [K+]o increases ion transport transiently, dependent on K+ channels.
dc.format.extent
dc.language.iso en
dc.relation.ispartofseries Investigative Ophthalmology and Visual Science; 61(6)
dc.rights info:eu-repo/semantics/openAccess
dc.subject Augnlækningar
dc.subject Ion transport
dc.subject Mouse
dc.subject NaK-ATPase
dc.subject Retinal pigment epithelium
dc.subject Ussing
dc.subject Ophthalmology
dc.subject Sensory Systems
dc.subject Cellular and Molecular Neuroscience
dc.title Mechanisms of Ion Transport across the Mouse Retinal Pigment Epithelium Measured in Vitro
dc.type /dk/atira/pure/researchoutput/researchoutputtypes/contributiontojournal/article
dc.description.version Peer reviewed
dc.identifier.pmid 32539134
dc.identifier.doi https://doi.org/10.1167/IOVS.61.6.31
dc.relation.url http://www.scopus.com/inward/record.url?scp=85086623966&partnerID=8YFLogxK
dc.contributor.department Faculty of Medicine
dc.contributor.department Other departments


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