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