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PMID:21566056

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Citation

He, B, Ebarasi, L, Hultenby, K, Tryggvason, K and Betsholtz, C (2011) Podocin-green fluorescence protein allows visualization and functional analysis of podocytes. J. Am. Soc. Nephrol. 22:1019-23

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.

Links

PubMed PMC3374364 Online version:10.1681/ASN.2010121291

Keywords

Animals; Animals, Genetically Modified; Green Fluorescent Proteins/genetics; Green Fluorescent Proteins/metabolism; Intracellular Signaling Peptides and Proteins/genetics; Intracellular Signaling Peptides and Proteins/metabolism; Kidney Glomerulus/cytology; Kidney Glomerulus/metabolism; Kidney Glomerulus/ultrastructure; Membrane Proteins/genetics; Membrane Proteins/metabolism; Microscopy, Electron, Scanning; Models, Animal; Podocytes/cytology; Podocytes/metabolism; Podocytes/ultrastructure; Zebrafish; Zebrafish Proteins/genetics; Zebrafish Proteins/metabolism

Significance

Annotations

Gene product Qualifier GO Term Evidence Code with/from Aspect Extension Notes Status

DANRE:A0A0R4IBP9

involved_in

GO:0032835: glomerulus development

ECO:0000315: mutant phenotype evidence used in manual assertion

ZFIN:ZDB-MRPHLNO-060612-1

P

Seeded From UniProt

complete

DANRE:I3VKD7

involved_in

GO:0032835: glomerulus development

ECO:0000315: mutant phenotype evidence used in manual assertion

ZFIN:ZDB-MRPHLNO-060612-1

P

Seeded From UniProt

complete

DANRE:Q1A5L1

involved_in

GO:0032835: glomerulus development

ECO:0000315: mutant phenotype evidence used in manual assertion

ZFIN:ZDB-MRPHLNO-060612-1

P

Seeded From UniProt

complete


See also

References

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