Abstract: FR-PO998

Development of Method for Assessing Chemical-Induced Toxicity against Kidney Cells by In Vivo Live Imaging Technique Using Nephron-Visualized-Transparent-Zebrafish

Session Information

Category: Bioengineering and Informatics

  • 101 Bioengineering and Informatics

Authors

  • Akiyama, Shin'ichi, Nagoya University Graduate School of Medicine, Nagoya, Japan
  • Maruyama, Shoichi, Nagoya University Graduate School of Medicine, Nagoya, Japan
Background

Zebrafish is a well-established vertebrate model for studying genetically higher levels of biological phenomena and it is suitable for the identification of toxic- or therapeutic-compounds on a larger scale. Larva of zebrafish are transparent, so they are often used for in vivo phenotype-based toxicological assay, but because the body of juvenile or adult fish body is not transparent, it is not suitable for that.
In this study, we tried to develop a new platform for assessing chemicals safety and toxicity against kidney by using a nephron-visualized-transparent-zebrafish.

Methods

We generated nephron-visualized-transparent zebrafish, which expressing green fluorescent protein (GFP) in glomeruli and proximal-mid tubule, by bioengineering technique using a promoter region of slc20a1a gene, a Gal4-UAS system, GFP cDNA and transparent zebrafish (Casper line). The larvae, juvenile and young adult of this fish were administrated a gentamicin, as positive control of nephrotoxicin, or several compounds by microinjection or exposure. Further, when evaluating protein leakage from the glomeruli, the dye-labeled dextran (MW, 70k) was administered by microinjection. At 24 or 48 hours post administration, we evaluated the survival, edema formation, form and number of nephron, and leakage of dextran.

Results

Live nephrons of the nephron visualized transparent zebrafish, could be clearly observed through the skin from outside the body from larva to young adult fish (2 months post-fertilization). In the fish treated with nephrotoxicin, edema increase, nephron loss, and dextran leakage were observed in a dose-dependent manner.

Conclusion

These results suggest that our in vivo assessing technique using the nephron-visualized-transparent-zebrafish have the potential to provide more easy and advanced platforms for assessing kidney health impacts of chemicals.

Funding

  • Government Support - Non-U.S.