Journal
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
Volume 23, Issue 23, Pages -Publisher
MDPI
DOI: 10.3390/ijms232315069
Keywords
zebrafish; trypsin digestion; retinal vasculature; hyperglycemia; blood-retinal barrier; tight junction
Funding
- Korea Institute of Oriental Medicine [K15270, KSN20213302]
- National Marine Biodiversity Institute of Korea - Ministry of Oceans and Fisheries [2022M00400, 20220027]
- National Research Foundation of Korea - Ministry of Education [2018R1A6A1A03025523]
- National Research Council of Science & Technology (NST), Republic of Korea [K15270] Funding Source: Korea Institute of Science & Technology Information (KISTI), National Science & Technology Information Service (NTIS)
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The evaluation of retinal vascular structures is crucial for studying ophthalmic diseases. This study introduces a rapid and convenient method to investigate the fine vessel structures in the intact retinal vasculature of adult zebrafish. The method allows for detailed examination of tight junction structures and identification of different cell types in the retinal vasculature. The findings also show the potential of using this method to study retinal vascular diseases in zebrafish models.
The evaluation of retinal vascular structures is important for analyzing various ophthalmic diseases. Conventional trypsin digestion was used for separating retinal vasculatures in mouse, rat, and other animal models; however, the trypsin method alone is technically difficult to perform and has not been reported in zebrafish to date. In this study, we introduced a rapid and convenient method that allows the investigation of fine vessel structures at a cellular level in the relatively intact retinal vasculature of adult zebrafish. Using an anti-ZO-1 antibody, tight junction structures in retinal vessels were examined in detail and several different cell types constituting blood vessels in arterial and capillary areas were identified. In addition, using cell type-specific antibodies, we identified smooth muscle cells, blood cells, and endothelial cells in the retinal vasculature. Finally, using the hyperglycemic model, we observed the dilation of retinal vessels, the downregulation of tight junction proteins, and the reduction in smooth muscle cells. Based on these results, we provide a rapid and convenient method for the study of retinal vasculature disease in the zebrafish animal model.
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