April 2014
Volume 55, Issue 13
Free
ARVO Annual Meeting Abstract  |   April 2014
Retinal Regeneration in Zebrafish: A New Model for Focal Injury to the Outer Retina
Author Affiliations & Notes
  • Alex Yuan
    Ophthalmology, Cleveland Clinic Foundation, Cleveland, OH
  • Brent A Bell
    Ophthalmology, Cleveland Clinic Foundation, Cleveland, OH
  • Rose M. DiCicco
    Ophthalmology, Cleveland Clinic Foundation, Cleveland, OH
  • Charlie Kaul
    Ophthalmology, Cleveland Clinic Foundation, Cleveland, OH
  • Joe G Hollyfield
    Ophthalmology, Cleveland Clinic Foundation, Cleveland, OH
  • Brian D Perkins
    Ophthalmology, Cleveland Clinic Foundation, Cleveland, OH
  • Bela Anand-Apte
    Ophthalmology, Cleveland Clinic Foundation, Cleveland, OH
  • Yuankai K Tao
    Ophthalmology, Cleveland Clinic Foundation, Cleveland, OH
  • Footnotes
    Commercial Relationships Alex Yuan, None; Brent Bell, None; Rose DiCicco, None; Charlie Kaul, None; Joe Hollyfield, None; Brian Perkins, None; Bela Anand-Apte, None; Yuankai Tao, None
  • Footnotes
    Support None
Investigative Ophthalmology & Visual Science April 2014, Vol.55, 1443. doi:
  • Views
  • Share
  • Tools
    • Alerts
      ×
      This feature is available to authenticated users only.
      Sign In or Create an Account ×
    • Get Citation

      Alex Yuan, Brent A Bell, Rose M. DiCicco, Charlie Kaul, Joe G Hollyfield, Brian D Perkins, Bela Anand-Apte, Yuankai K Tao; Retinal Regeneration in Zebrafish: A New Model for Focal Injury to the Outer Retina. Invest. Ophthalmol. Vis. Sci. 2014;55(13):1443.

      Download citation file:


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

      ×
  • Supplements
Abstract
 
Purpose
 

To develop a novel laser-induced injury model to study retinal regeneration in zebrafish.

 
Methods
 

Adult zebrafish were imaged by optical coherence tomography (OCT) and confocal scanning laser ophthalmoscopy (cSLO) in room air through a contact lens. Using a custom designed beam splitter, 532nm laser photocoagulation was applied using the OCT C-scan image for targeting. Calibrated laser spots of 42-47mW were delivered to the retina. At multiple intervals post-lesion, fish were re-imaged using both OCT and cSLO to follow the progression of each laser burn. Histologic sections were performed at various intervals to monitor the injury response. Lesion size was measured using cSLO images and ImageJ software.

 
Results
 

Retinal lesions were visible by OCT and cSLO immediately after laser application. Early OCT changes include an increase in hyper-reflectance and loss of the outer retina banding pattern. Lesions were best detected using infrared darkfield imaging on cSLO, but they were also visible with autofluorescence imaging. The average lesion size on day 1 was 26057 ± 397μm2. After 1 week the lesion size and intensity began to decrease and at 15 weeks, lesions were barely detectable above background and the average size was 9404μm2. Histologic sections showed focal areas of damage localized to the outer retina. At 15 weeks, the damaged retina had regenerated and re-established a laminated structure. However, subtle changes can still be detected by cSLO and OCT imaging.

 
Conclusions
 

OCT guided laser photocoagulation can be used to induce focal retinal injury in zebrafish. Histologic analysis confirmed focal and selective damage to the outer retina. By week 15, the normal lamination and architecture of the retina was re-established. This laser model allows for focal destruction of the retina, leaving the surrounding retina intact, allowing for improved study of cellular proliferation, migration, and differentiation during the regenerative process in zebrafish.

 
 
Lesion size decreases over time. (Top) Plot of average lesion size over time. (Bottom) Representative cSLO images 0-9 weeks after laser. An outline of the superior lesion is shown in black.
 
Lesion size decreases over time. (Top) Plot of average lesion size over time. (Bottom) Representative cSLO images 0-9 weeks after laser. An outline of the superior lesion is shown in black.
 
 
Comparison of lesions 2 days and 15 weeks post-laser. (Top) H&E, (middle) OCT, (bottom) cSLO. Each set of images were obtained from a single animal. Black arrowheads point to the corresponding lesions.
 
Comparison of lesions 2 days and 15 weeks post-laser. (Top) H&E, (middle) OCT, (bottom) cSLO. Each set of images were obtained from a single animal. Black arrowheads point to the corresponding lesions.
 
Keywords: 687 regeneration • 578 laser • 688 retina  
×
×

This PDF is available to Subscribers Only

Sign in or purchase a subscription to access this content. ×

You must be signed into an individual account to use this feature.

×