The Apical Localization of Na+, K+-ATPase in Cultured Human Retinal Pigment Epithelial Cells Depends on Expression of the β2 Subunit
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Na+, K+-ATPase, or the Na+ pump, is a key component in the maintenance of the epithelial phenotype. In most epithelia, the pump is located in the basolateral domain. Studies from our laboratory have shown that the β1 subunit of Na+, K+-ATPase plays an important role in this mechanism because homotypic β1-β1 interactions between neighboring cells stabilize the pump in the lateral membrane. However, in the retinal pigment epithelium (RPE), the Na+ pump is located in the apical domain. The mechanism of polarization in this epithelium is unclear. We hypothesized that the apical polarization of the pump in RPE cells depends on the expression of its β2 subunit. ARPE-19 cells cultured for up to 8 weeks on inserts did not polarize, and Na+, K+-ATPase was expressed in the basolateral membrane. In the presence of insulin, transferrin and selenic acid (ITS), ARPE-19 cells cultured for 4 weeks acquired an RPE phenotype, and the Na+ pump was visible in the apical domain. Under these conditions, Western blot analysis was employed to detect the β2 isoform and immunofluorescence analysis revealed an apparent apical distribution of the β2 subunit. qPCR results showed a time-dependent increase in the level of β2 isoform mRNA, suggesting regulation at the transcriptional level. Moreover, silencing the expression of the β2 isoform in ARPE-19 cells resulted in a decrease in the apical localization of the pump, as assessed by the mislocalization of the α2 subunit in that domain. Our results demonstrate that the apical polarization of Na+, K+-ATPase in RPE cells depends on the expression of the β2 subunit.
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The β2 subunit of the Na+/K+-ATPase localizes to the apical surface of RPE cells, rather than the basolateral surface as in most other epithelia.
“A striking example of this is expression of the β2 subunit of the Na+/K+-ATPase, which results in the localization of this pump to the apical surface, rather than the basolateral surface, as found in most other epithelia (Lobato-Alvarez et al., 2016).”
From The cell biology of the retinal pigment epithelium (2020) · cites this paper as Lobato-Alvarez et al., 2016
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- Authors
- Lobato-Álvarez, J. A., Roldán, M. L., López-Murillo, T. D. C., González-Ramírez, R., Bonilla-Delgado, J., & Shoshani, L.
- Affiliations
- Centro de Investigación y de Estudios Avanzados del Instituto Politécnico Nacional (MX)
- Corresponding author
- Shoshani, Liora
- ORCID
- Shoshani, Liora
- Venue
- Frontiers in Physiology
- Publisher
- Frontiers Media SA
- Published
- 2016-10-07
- Volume
- 7
- Pages
- 450-450
- ISSN
- 1664-042X (ISSN-L)
- PMC
- PMC5054689
- OpenAlex
- W2528637216
- Cited by
- 27
- References
- 74
- Citations by year
- 2026: 1 · 2025: 5 · 2024: 4 · 2023: 3 · 2022: 3 · 2021: 4 · 2020: 4 · 2019: 1 · 2018: 1 · 2017: 1
- FWCI
- 0.52
- Type
- journal article
- Language
- en
- Open access
- gold
- doi.org
- Licence
- cc-by
- Locations
- Frontiers in Physiology (publishedVersion, cc-by)
- Europe PMC (PubMed Central) (publishedVersion, cc-by)
- PubMed Central (publishedVersion, cc-by)
- PubMed (publishedVersion)
- DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
- Funders
- Consejo Nacional de Ciencia y Tecnología (127460)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Ion Transport and Channel Regulation
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Retinal Development and Disorders
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › ATP Synthase and ATPases Research
- Keywords
- Na+/K+-ATPase, beta2 subunit, retinal pigment epithelium, apical membrane localization, epithelial polarization, ARPE-19 cells, insulin, transferrin, selenic acid, beta1 subunit, alpha2 subunit, transcriptional regulation
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