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Directional Exosome Proteomes Reflect Polarity-Specific Functions in Retinal Pigmented Epithelium Monolayers.

Publication ,  Journal Article
Klingeborn, M; Dismuke, WM; Skiba, NP; Kelly, U; Stamer, WD; Bowes Rickman, C
Published in: Sci Rep
July 7, 2017

The retinal pigmented epithelium (RPE) forms the outer blood-retinal barrier in the eye and its polarity is responsible for directional secretion and uptake of proteins, lipoprotein particles and extracellular vesicles (EVs). Such a secretional division dictates directed interactions between the systemic circulation (basolateral) and the retina (apical). Our goal is to define the polarized proteomes and physical characteristics of EVs released from the RPE. Primary cultures of porcine RPE cells were differentiated into polarized RPE monolayers on permeable supports. EVs were isolated from media bathing either apical or basolateral RPE surfaces, and two subpopulations of small EVs including exosomes, and dense EVs, were purified and processed for proteomic profiling. In parallel, EV size distribution and concentration were determined. Using protein correlation profiling mass spectrometry, a total of 631 proteins were identified in exosome preparations, 299 of which were uniquely released apically, and 94 uniquely released basolaterally. Selected proteins were validated by Western blot. The proteomes of these exosome and dense EVs preparations suggest that epithelial polarity impacts directional release. These data serve as a foundation for comparative studies aimed at elucidating the role of exosomes in the molecular pathophysiology of retinal diseases and help identify potential therapeutic targets and biomarkers.

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Published In

Sci Rep

DOI

EISSN

2045-2322

Publication Date

July 7, 2017

Volume

7

Issue

1

Start / End Page

4901

Location

England

Related Subject Headings

  • Swine
  • Retinal Pigment Epithelium
  • Proteome
  • Primary Cell Culture
  • Molecular Sequence Annotation
  • Mass Spectrometry
  • Gene Ontology
  • Gene Expression Regulation
  • Gene Expression Profiling
  • Eye Proteins
 

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Klingeborn, M., Dismuke, W. M., Skiba, N. P., Kelly, U., Stamer, W. D., & Bowes Rickman, C. (2017). Directional Exosome Proteomes Reflect Polarity-Specific Functions in Retinal Pigmented Epithelium Monolayers. Sci Rep, 7(1), 4901. https://doi.org/10.1038/s41598-017-05102-9
Klingeborn, Mikael, W Michael Dismuke, Nikolai P. Skiba, Una Kelly, W Daniel Stamer, and Catherine Bowes Rickman. “Directional Exosome Proteomes Reflect Polarity-Specific Functions in Retinal Pigmented Epithelium Monolayers.Sci Rep 7, no. 1 (July 7, 2017): 4901. https://doi.org/10.1038/s41598-017-05102-9.
Klingeborn M, Dismuke WM, Skiba NP, Kelly U, Stamer WD, Bowes Rickman C. Directional Exosome Proteomes Reflect Polarity-Specific Functions in Retinal Pigmented Epithelium Monolayers. Sci Rep. 2017 Jul 7;7(1):4901.
Klingeborn, Mikael, et al. “Directional Exosome Proteomes Reflect Polarity-Specific Functions in Retinal Pigmented Epithelium Monolayers.Sci Rep, vol. 7, no. 1, July 2017, p. 4901. Pubmed, doi:10.1038/s41598-017-05102-9.
Klingeborn M, Dismuke WM, Skiba NP, Kelly U, Stamer WD, Bowes Rickman C. Directional Exosome Proteomes Reflect Polarity-Specific Functions in Retinal Pigmented Epithelium Monolayers. Sci Rep. 2017 Jul 7;7(1):4901.

Published In

Sci Rep

DOI

EISSN

2045-2322

Publication Date

July 7, 2017

Volume

7

Issue

1

Start / End Page

4901

Location

England

Related Subject Headings

  • Swine
  • Retinal Pigment Epithelium
  • Proteome
  • Primary Cell Culture
  • Molecular Sequence Annotation
  • Mass Spectrometry
  • Gene Ontology
  • Gene Expression Regulation
  • Gene Expression Profiling
  • Eye Proteins