June 2017
Volume 58, Issue 8
Open Access
ARVO Annual Meeting Abstract  |   June 2017
Comparison of zebrafish retinal proteins during experimental degeneration and regeneration using quantitative proteomics
Author Affiliations & Notes
  • Karen Eastlake
    National Institute for Health Research (NIHR) Biomedical Research Centre at Moorfields Eye Hospital NHS Foundation Trust and UCL Institute of Ophthalmology, London, United Kingdom
  • Wendy Heywood
    Centre for Translational Omics, UCL Great Ormond Street Institute of Child Health, London, United Kingdom
  • Dhani C Tracey-White
    National Institute for Health Research (NIHR) Biomedical Research Centre at Moorfields Eye Hospital NHS Foundation Trust and UCL Institute of Ophthalmology, London, United Kingdom
  • Erika Aquino
    National Institute for Health Research (NIHR) Biomedical Research Centre at Moorfields Eye Hospital NHS Foundation Trust and UCL Institute of Ophthalmology, London, United Kingdom
  • Emily Bliss
    Centre for Translational Omics, UCL Great Ormond Street Institute of Child Health, London, United Kingdom
  • Gerardo Vasta
    Department of Microbiology and Immunology, University of Maryland School of Medicine and IMET, Baltimore, Maryland, United States
  • Kevin Mills
    Centre for Translational Omics, UCL Great Ormond Street Institute of Child Health, London, United Kingdom
  • Peng Tee Khaw
    National Institute for Health Research (NIHR) Biomedical Research Centre at Moorfields Eye Hospital NHS Foundation Trust and UCL Institute of Ophthalmology, London, United Kingdom
  • Mariya Moosajee
    National Institute for Health Research (NIHR) Biomedical Research Centre at Moorfields Eye Hospital NHS Foundation Trust and UCL Institute of Ophthalmology, London, United Kingdom
  • G. Astrid Limb
    National Institute for Health Research (NIHR) Biomedical Research Centre at Moorfields Eye Hospital NHS Foundation Trust and UCL Institute of Ophthalmology, London, United Kingdom
  • Footnotes
    Commercial Relationships   Karen Eastlake, None; Wendy Heywood, None; Dhani Tracey-White, None; Erika Aquino, None; Emily Bliss, None; Gerardo Vasta, None; Kevin Mills, None; Peng Khaw, None; Mariya Moosajee, None; G. Astrid Limb, None
  • Footnotes
    Support  Fight for Sight; Moorfields Eye charity; MRC
Investigative Ophthalmology & Visual Science June 2017, Vol.58, 365. doi:
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      Karen Eastlake, Wendy Heywood, Dhani C Tracey-White, Erika Aquino, Emily Bliss, Gerardo Vasta, Kevin Mills, Peng Tee Khaw, Mariya Moosajee, G. Astrid Limb; Comparison of zebrafish retinal proteins during experimental degeneration and regeneration using quantitative proteomics. Invest. Ophthalmol. Vis. Sci. 2017;58(8):365.

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

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Abstract

Purpose : Zebrafish have the ability to regenerate retina after injury. Although many studies have investigated the retinal gene profile in this species during regeneration, understanding mechanisms involved in this process is far from complete as few studies have investigated the biological function at protein level. To address this we compared the proteomic profile of the zebrafish retina after injury and upon regeneration.

Methods : Adult longfin wildtype zebrafish eyes were injected with 200μM Ouabain to induce degeneration. Protein isolated from retina excised at 0, 3 and 18 days after injection -corresponding to normal, degenerated and regenerating states- were analysed using two-dimensional difference gel electrophoresis (2D-DIGE) and label-free quantitative proteomics using quadrupole time of flight (QToF) LC-MS/MS. Protein identification and bioinformatics analysis were performed using Protein Lynx Global Server (PLGS), Non-Linear dynamics Progenesis software and online Panther gene ontology database. Results were compared to RT-PCR and western blots from excised retina

Results : Across all specimens, 2042 proteins were identified. Proteins showing the highest fold upregulation in the degenerated retina included fibrinogen gamma polypeptide, apolipoproteins A-Ib and A-II, galectin-1 and vitellogenin-6. In addition, gene ontology analysis indicates reduced metabolic processing, and an increase in fibrin clot formation. In the regenerating retina, cytoskeleton and membrane transport proteins were considerably altered, with the highest fold upregulation observed for tubulin beta 2A, histone H2B and brain type fatty acid binding protein. RT-PCR and western blot analysis confirmed that galectin-1 expression was significantly upregulated in the degenerated retina as compared to control or regenerating retina.

Conclusions : Investigations on the role of key proteins identified in the regenerating zebrafish retina may aid not only to understand mechanisms that prevent regeneration of the adult human retina, but to the design of new approaches to stimulate endogenous repair mechanisms following retinal degenerative disease.

This is an abstract that was submitted for the 2017 ARVO Annual Meeting, held in Baltimore, MD, May 7-11, 2017.

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