“Zebrafishing” for Novel Genes Relevant to the Glomerular Filtration Barrier

Figure 4

Eye assay and dot blot for measuring glomerular filter integrity using transgenic l-fabp:DBP-eGFP zebrafish. The l-fabp:DBP-eGFP transgenic zebrafish produces a green-fluorescent plasma protein. The transgene expression is driven by the fabp-liver promoter and leads to expression of a vitamin D binding protein fused with eGFP. The promoter becomes active at 2 days postfertilization that leads to the production and accumulation of fluorescent plasma protein that can be monitored over the retinal vessel plexus. If a morpholino injection leads to a compromise of the glomerular filtration barrier, fluorescence accumulation measured over the retinal plexus does not occur, and the eGFP that is lost via the kidney can be detected in the fish water, for example, using a dot blot approach.