|ZFIN ID: ZDB-PUB-110524-19|
Podocin-Green Fluorescence Protein Allows Visualization and Functional Analysis of Podocytes
He, B., Ebarasi, L., Hultenby, K., Tryggvason, K., and Betsholtz, C.
|Source:||Journal of the American Society of Nephrology : JASN 22(6): 1019-1023 (Journal)|
|Registered Authors:||Betsholtz, Christer, Ebarasi, Lwaki, He, Bing, Tryggvason, Karlynn|
|PubMed:||21566056 Full text @ J. Am. Soc. Nephrol.|
He, B., Ebarasi, L., Hultenby, K., Tryggvason, K., and Betsholtz, C. (2011) Podocin-Green Fluorescence Protein Allows Visualization and Functional Analysis of Podocytes. Journal of the American Society of Nephrology : JASN. 22(6):1019-1023.
ABSTRACTPodocytes 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.