ＭＺＴ（株）ゼブラフィッシュ創薬研究所

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high-throughput individual zebrafish screening

野本毅1,2、清水陽嘉1,2、寺見文宏1,2、森葵泉1,2、松岡さおり1,2、櫛田友紀1,2、西野加奈子1,2、○田中利男1,2
1三重大学大学院医学系研究科システムズ薬理学
2三重大学メディカルゼブラフィッシュ研究センター

Zebrafish have been extensively used for nephrotoxicity screening worldwide, because of their 84% homology to the human pathological genome, which suggests similarity to humans in toxicity mechanisms, and their whole-body transparency, which allows direct analysis of renal morphology and renal function. In addition, zebrafish is the most suitable vertebrate for in vivo nephrotoxicity high-throughput screening due to its high fecundity and rapid organogenesis. However, conventional methods were not sensitive enough for early diagnosis. Therefore, it was decided to apply intravenous fluorescent labeled dextran of one molecular weight for nephrotoxic proteinuria quantification. But it is not an endogenous protein and has only one molecular weight. Therefore, the determination of nephrotoxic proteinuria by transgenic zebrafish of plasma proteins (1/2vdbp-mCherry:50kDa, NL-D3:35.5kDa) was reported. However, in these cases, the molecular weight was still one of a kind, and the inability to discriminate between glomerular and tubular functions in nephrotoxicity remained an issue. Therefore, in 2013 (ACS Chem Neurosci. 4:1183-93), we independently developed a novel fluorescent dye ZMB741 that rapidly binds to all molecular weight plasma proteins in vivo. We have established a protocol for in vivo quantification of nephrotoxic proteinuria in individual zebrafish. As a result, we report the creation of an in vivo high-throughput nephrotoxicity screening system based on a completely new principle that can discriminate between glomerular and tubular function in nephrotoxicity.

ＭＺＴ（株）ゼブラフィッシュ創薬研究所

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high-throughput individual zebrafish screening

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