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Jing Xie, Alecia Cutler, Mariya Ali, Bela Anand-Apte; Retinoic acid signaling has an important role in development and maintenance of the blood retinal barrier. Invest. Ophthalmol. Vis. Sci. 2013;54(15):3746.
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In our previous work, we have demonstrated that the blood-retinal barrier (BRB) is established in zebrafish at 3 days postfertilization (dpf), and integrity of the BRB can be visualized in vivo using a transgenic zebrafish line, Tg(l-fabp:DBP-EGFP). The purpose of this study is to determine whether retinoic acid (RA) signaling is important for the BRB.
The l-fabp:DBP-EGFP embryos were treated with 5-20 uM DEAB or BMS 493, two specific inhibitors of the RA signaling, with or without all-trans retinoic acid (ATRA) from 2 dpf. From 5 to 7 dpf, leakage of the EGFP was detected under a fluorescent microscope. The RA signaling was also knocked down in the Tg(hsp:Cyp26a1) zebrafish to test its function the BRB.
The treatment of DEAB or BMS 493, results in non-functional BRB in 63-95% fish, while 0.5 uM ATRA can effectively rescue the BRB development. Similarly, the BRB can be disrupted in the Tg(hsp:Cyp26a1) zebrafish, either at embryonic or larval stage. The expression of the claudin-5 and ZO-1, two markers of the BRB, is also decreased in the fish treated with the inhibitors.
These results demonstrate that the RA signaling is essential in development and maintenance of the zebrafish BRB.
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