Dataset Information


Podocin-green fluorescence protein allows visualization and functional analysis of podocytes.

ABSTRACT: Podocytes do not remain fully differentiated when cultured, and they are difficult to image in vivo, making the study of podocyte biology challenging. Zebrafish embryos are transparent and develop a single, midline, pronephric glomerulus accessible for imaging and systematic functional analysis. Here, we describe a transgenic zebrafish line that expresses green fluorescence protein (GFP) from the zebrafish podocin promoter. The line recapitulates the endogenous pronephric podocin expression pattern, showing GFP expression exclusively in podocytes starting 2 days postfertilization. Using the podocyte GFP signal as a guide for dissection, we examined the pronephric glomerulus by scanning electron microscopy; the surface ultrastructure exhibited fine, interdigitating podocyte foot processes surrounding glomerular capillaries. To determine whether the GFP signal could serve as a direct readout of developmental abnormalities or injury to the glomerulus, we knocked down the podocyte-associated protein crb2b; this led to a loss of GFP signal. Thus, podocin-GFP zebrafish provide a model for ultrastructural studies and in vivo visualization and functional analysis of glomerular podocytes. This model should also be useful for high-throughput genetic or chemical analysis of glomerular development and function.

PROVIDER: S-EPMC3374364 | BioStudies |

REPOSITORIES: biostudies

Similar Datasets

| S-EPMC2836015 | BioStudies
| S-EPMC3819108 | BioStudies
| S-EPMC3358760 | BioStudies
| S-EPMC5696060 | BioStudies
| S-EPMC5639718 | BioStudies
| S-EPMC3396653 | BioStudies
| S-EPMC343810 | BioStudies
| S-EPMC3884556 | BioStudies
| S-EPMC6035194 | BioStudies
| S-EPMC4814194 | BioStudies