July 2018
Volume 59, Issue 9
Open Access
ARVO Annual Meeting Abstract  |   July 2018
Voltage-gated sodium channels in retinal pigment epithelium – a real deal or an artifact?
Author Affiliations & Notes
  • Julia Kristiina Johansson
    Faculty of Biomedical Sciences and Engineering, Tampere University of Technology, Tampere, Finland
    BioMediTech, Tampere University of Technology, Tampere, Finland
  • Teemu Ihalainen
    Faculty of Medicine and Life Sciences, University of Tampere, Tampere, Finland
    BioMediTech, University of Tampere, Tampere, Finland
  • Heli Skottman
    Faculty of Medicine and Life Sciences, University of Tampere, Tampere, Finland
    BioMediTech, University of Tampere, Tampere, Finland
  • Soile Nymark
    Faculty of Biomedical Sciences and Engineering, Tampere University of Technology, Tampere, Finland
    BioMediTech, Tampere University of Technology, Tampere, Finland
  • Footnotes
    Commercial Relationships   Julia Johansson, None; Teemu Ihalainen, None; Heli Skottman, None; Soile Nymark, None
  • Footnotes
    Support  Emil Aaltonen Foundation, Academy of Finland Grant 294054
Investigative Ophthalmology & Visual Science July 2018, Vol.59, 1189. doi:
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      Julia Kristiina Johansson, Teemu Ihalainen, Heli Skottman, Soile Nymark; Voltage-gated sodium channels in retinal pigment epithelium – a real deal or an artifact?. Invest. Ophthalmol. Vis. Sci. 2018;59(9):1189.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose : Cells of retinal pigment epithelium (RPE) have been thought to undergo epithelial to neuronal transition by expressing voltage-gated sodium channels (Nav) in culture. Because Nav currents have not been recorded ex vivo, their presence in vitro is not considered to reflect the physiology of the native RPE. We revisited this question by investigating the presence of the Nav channels in mouse and human embryonic stem cell (hESC) derived RPE. In this study, the plausible roles of Nav channels in RPE were studied during development and in phagocytosis of the photoreceptor outer segments (POS).

Methods : Patch clamp recordings were conducted from intact monolayers of hESC-RPE and freshly dissociated cells at various stages of RPE development with and without the presence of Nav channel blockers. Immunolabelling from both hESC-derived and mouse RPE was used to investigate the channel subtype distribution and localization as well as their changes during development. The phagocytosis was assayed in vitro by incubating the purified porcine outer segments on hESC-RPE and ex vivo by preparing the mouse eyes with the retina intact at various timepoints of the circadian cycle.

Results : Recordings from the monolayer showed the presence of NaV current in hESC-RPE that was inhibited by TTX and QX-314. The current was mainly carried by subtypes Nav1.4-Nav1.6 and Nav1.8, and these channels were observed throughout the development. The set of channel subtypes we identified in the RPE were not characteristic of the neural retina and, notably, the main neuronal subtypes Nav1.1 and Nav1.2 were not observed in mature RPE. Interestingly, the currents identified in isolated RPE cells were infrequent and had reduced amplitudes. The immunolabeling indicated that dissociation causes the channels to concentrate to cellular infoldings where they might not be accessible to pass ionic currents. Finally, the phagocytosis assays revealed a possible role for Nav1.4 and Nav1.8 in POS uptake and indicated the accumulation of these channels to the phagosomes.

Conclusions : Our data strongly indicates that NaV channels are present in RPE throughout development and that they are relevant for its physiology. The findings from mouse RPE challenge the notion that these channels appear because of the epithelial to neuronal transition in culture.

This is an abstract that was submitted for the 2018 ARVO Annual Meeting, held in Honolulu, Hawaii, April 29 - May 3, 2018.

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