PUBLICATION
Podocin-Green Fluorescence Protein Allows Visualization and Functional Analysis of Podocytes
- Authors
- He, B., Ebarasi, L., Hultenby, K., Tryggvason, K., and Betsholtz, C.
- ID
- ZDB-PUB-110524-19
- Date
- 2011
- Source
- Journal of the American Society of Nephrology : JASN 22(6): 1019-1023 (Journal)
- Registered Authors
- Betsholtz, Christer, Ebarasi, Lwaki, He, Bing, Tryggvason, Karlynn
- Keywords
- none
- MeSH Terms
-
- Models, Animal
- Animals
- Kidney Glomerulus/cytology
- Kidney Glomerulus/metabolism
- Kidney Glomerulus/ultrastructure
- Membrane Proteins/genetics
- Membrane Proteins/metabolism*
- Intracellular Signaling Peptides and Proteins/genetics
- Intracellular Signaling Peptides and Proteins/metabolism*
- Microscopy, Electron, Scanning
- Animals, Genetically Modified
- Zebrafish Proteins/genetics
- Zebrafish Proteins/metabolism
- Podocytes/cytology*
- Podocytes/metabolism*
- Podocytes/ultrastructure
- Zebrafish
- Green Fluorescent Proteins/genetics
- Green Fluorescent Proteins/metabolism*
- PubMed
- 21566056 Full text @ J. Am. Soc. Nephrol.
Citation
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.
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.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping