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PMID:11786407

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Citation

Roselli, S, Gribouval, O, Boute, N, Sich, M, Benessy, F, Attié, T, Gubler, MC and Antignac, C (2002) Podocin localizes in the kidney to the slit diaphragm area. Am. J. Pathol. 160:131-9

Abstract

We recently cloned a novel gene, NPHS2, involved in autosomal recessive steroid-resistant nephrotic syndrome. This gene encodes a novel podocyte protein, podocin. Given its similarity with the stomatin family proteins, podocin is predicted to be an integral membrane protein with a single membrane domain forming a hairpin-like structure placing both N- and C-termini in the cytosol. Here, we show by in situ hybridization, that during development, the NPHS2 transcript is first expressed in mesonephric podocytes from the S-shaped body and, later, in the metanephric kidney, in the future podocytes at the late S-shaped body stage. In the mature kidney, NPHS2 is exclusively expressed in the podocytes of mature glomeruli. We generated rabbit polyclonal antibodies against fusion proteins derived from the N- and the C-terminal regions of podocin which detected a single band of 49-kd in transfected HEK293 cell lysates by immunoprecipitation and Western blotting. By immunohistology, podocin was detected in podocytes from the early capillary loop stage in the developing nephrons, and at the basal pole, along the GBM, in mature glomeruli. By electron microscopy, we demonstrate that podocin is facing the slit diaphragm with its two ends in the cytoplasm of the foot processes, in agreement with its predicted structure. Our results suggest that podocin could serve to anchor directly or indirectly components of the slit diaphragm to the cytoskeleton.

Links

PubMed PMC1867125 Online version:10.1016/S0002-9440(10)64357-X

Keywords

Cell Line; Fetus; Fluorescent Antibody Technique; Humans; Intercellular Junctions/metabolism; Intercellular Junctions/ultrastructure; Intracellular Signaling Peptides and Proteins; Kidney/metabolism; Kidney Glomerulus/metabolism; Kidney Glomerulus/ultrastructure; Membrane Proteins/genetics; Membrane Proteins/metabolism; Microscopy, Confocal; Microscopy, Immunoelectron; RNA, Messenger/metabolism; Tissue Distribution

Significance

Annotations

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

MOUSE:PODO

located_in

GO:0031235: intrinsic component of the cytoplasmic side of the plasma membrane

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

MOUSE:PODO

colocalizes_with

GO:0036057: slit diaphragm

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

HUMAN:PODO

located_in

GO:0031235: intrinsic component of the cytoplasmic side of the plasma membrane

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

HUMAN:PODO

colocalizes_with

GO:0036057: slit diaphragm

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

HUMAN:PODO

involved_in

GO:0072249: metanephric glomerular visceral epithelial cell development

ECO:0000270: expression pattern evidence used in manual assertion

P

Seeded From UniProt

complete


See also

References

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