June 2013
Volume 54, Issue 15
Free
ARVO Annual Meeting Abstract  |   June 2013
The Small Heat Shock Protein αB-crystallin is required for Exosome Biogenesis in Adult Human Retinal Pigment Epithelial Cells
Author Affiliations & Notes
  • Suraj Bhat
    Ophthalmology, Jules Stein Eye Institute UCLA, Los Angeles, CA
    Molecular Biology Institute, Los Angeles, CA
  • Rajendra Gangalum
    Ophthalmology, Jules Stein Eye Institute UCLA, Los Angeles, CA
  • Ishanee Dighe
    Ophthalmology, Jules Stein Eye Institute UCLA, Los Angeles, CA
  • Ivo Atanasov
    California Nanosystems Institute @ UCLA, Los Angeles, CA
  • Z Hong Zhou
    California Nanosystems Institute @ UCLA, Los Angeles, CA
  • Larry David
    Oregon Health and Sciences University, Potland, OR
  • Footnotes
    Commercial Relationships Suraj Bhat, None; Rajendra Gangalum, None; Ishanee Dighe, None; Ivo Atanasov, None; Z Hong Zhou, None; Larry David, None
  • Footnotes
    Support None
Investigative Ophthalmology & Visual Science June 2013, Vol.54, 1202. doi:
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      Suraj Bhat, Rajendra Gangalum, Ishanee Dighe, Ivo Atanasov, Z Hong Zhou, Larry David; The Small Heat Shock Protein αB-crystallin is required for Exosome Biogenesis in Adult Human Retinal Pigment Epithelial Cells. Invest. Ophthalmol. Vis. Sci. 2013;54(15):1202.

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

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Abstract

Purpose: The expression of the small heat shock protein αB-crystallin is developmentally controlled and is known to be associated with a myriad of neuro-degenerations including Alzheimer’s disease, multiple sclerosis and age-related macular degeneration (AMD). We have previously shown that αB-crystallin is a Golgi membrane-associated protein, which is secreted from adult human retinal pigment epithelial cell (ARPE19) via exosomes (Gangalum et al., J Biol Chem. 286:3261-9, 2011). In this investigation we have probed the relevance of αB-crystallin to RPE function and intercellular communication via exosomes.

Methods: Employing shRNA technologies to inhibit specific gene expression we have generated stably transfected ARPE19 cells lines in which αB-crystallin expression is variably inhibited. We have used Fluorescence activated Cell sorting analyses, immunofluorescence, electron microscopy and biochemical characterizations and polarized cell cultures.

Results: Analyses of various cell lines expressing differential amounts of αB-crystallin protein show that absence of αB-crystallin protein has a significant impact on exosome biogenesis in human RPE cells in culture. Examination of various exosome markers and their status reveals that specific stages of exosome biogenesis are interfered with in the absence of this small heat shock protein.

Conclusions: Based on known cell-type-specificity of the molecular cargo of the exosomes these data indicate that cell-type specific proteins such as αB-crystallin may control intercellular communication in a cell-type specific fashion (and therefore, RPE physiology) by regulating exosome biogenesis.

Keywords: 412 age-related macular degeneration • 701 retinal pigment epithelium • 447 cell-cell communication  
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