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E. Yang, J. H. Kim, K.-W. Kim, Y. S. Yu, J. H. Kim; Investigation of Barrier Characteristics of Zebrafish Retinal Vessels. Invest. Ophthalmol. Vis. Sci. 2010;51(13):43.
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© ARVO (1962-2015); The Authors (2016-present)
Esther Yang1, Jin Hyoung Kim1, Kyu-Won Kim2, Young Suk Yu1, Jeong Hun Kim11Fight against Angiogenesis-Related Blindness (FARB) Laboratory, Department of Ophthalmology, College of Medicine, Seoul National University & Seoul Artificial Eye Center Clinical Research Institute, Seoul National University Hospital, Seoul, Korea; 2NeuroVascular Coordination Research Center, College of Pharmacy and Research Institute of Pharmaceutical Sciences, Seoul National University, Seoul, Korea
To investigate barrier characteristics of zebrafish retinal vessels.
Materials and Methods: :
Tg(fli1:EGFP) zebrafish, which expresses enhanced green fluorescent protein (EGFP) in all blood vessels throughout embryogenesis, were sacrificed at 5 and 7 days post-fertilization (dpf). Immunofluorescence staining of GFAP (astrocyte marker), NG2 (pericyte marker), and tight junction proteins (ZO-1, Occludin, and Claudin-5) was done for evaluation of barrier characteristics of retinal vessels. Vascular permeability was assessed in the retina and brain after injection of FITC-dextran (2000kDa), Rhodamin-dextran (10kDa), and DAPI (350Da) tracers.
With staining of GFAP and NG2, only NG2 was found to be co-localized with EGFP- expressing endothelial cells. ZO-1, Occludin, and Claudin-5 were all expressed in the zebrafish retina, but only Claudin-5 was expressed in the EGFP-expressing endothelial cells. In the assessment of vascular permeability with FITC-dextran, Rhodamin-dextran, and DAPI, retinal vessels were found to be permeable to all tracers, while brain vessels were impermeable to even DAPI.
Our results show that the zebrafish retinal vessel at 5 to 7 dpf was found to be lack of barrier characteristics, different from the brain vessel.
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