FIGURE 9.

Zebrafish injected with TAT-Neph1CD were resistant to PAN- and Ang II-induced injury. A, PAN (25 mg/ml) was co-injected with 0.25 mg/ml of either TAT-GFP (control) or TAT-Neph1CD (test) in 72-hpf zebrafish via the common cardinal vein, and the development of pericardial edema was monitored at 96 hpf. Significant increase in pericardial edema and curved body shape was noted in the control, whereas the test zebrafish had minimal or no edema, and their body shapes were grossly normal. B, quantitative analysis from three independent experiments suggested about 40% reduction of PAN-induced edema in test zebrafish as compared with the control. C and D, embryos either injected with 0.25 mg/ml of either TAT-GFP (control) or TAT-Neph1CD (test) were grown in the E3 medium containing adriamycin (30.3 mg/liter). The phenotype (pericardial edema an indication of renal abnormality) was observed at 96 hpf (more than 90% embryos develop edema in this model). However, the number of zebrafish embryos with pericardial edema was significantly reduced with TAT-Neph1CD injection. D, quantitative analysis further suggested a 25% decrease in pericardial edema in embryos injected with TAT-Neph1CD when compared with the embryos injected with TAT-GFP (p < 0.05). E, histological analysis of sections from 72 hpf of TAT-GFP-injected embryos treated with adriamycin and stained with hematoxylin and eosin showed prominent dilation of the pronephric tubules, whereas this phenotype was absent in TAT-Neph1CD-injected embryos treated with adriamycin. F, schematic representation of the in vivo filtration assay using transgenic zebrafish expressing GFP-VDBP from liver. Zebrafish injected with TAT-Neph1CD and uninjected were treated with adriamycin to induce glomerular injury, and the excreted GFP-VDBP was estimated in the medium. G, zebrafish embryos injected with TAT-Neph1CD showed a measurable reduction (∼30% with a p value of <0.05) in the excreted GFP-VDBP in the medium when compared with the control embryos.