Abstract
Podocytes are specialized epithelial cells of the kidney blood filtration barrier that contribute to permselectivity via a series of interdigitating actin-rich foot processes. Positioned between adjacent projections is a unique cell junction known as the slit diaphragm, which is physically connected to the actin cytoskeleton via the transmembrane protein nephrin. Evidence indicates that tyrosine phosphorylation of the intracellular tail of nephrin initiates signaling events, including recruitment of cytoplasmic adaptor proteins Nck1 andNck2 that regulate actin cytoskeletal dynamics. Nephrin tyrosine phosphorylation is altered in human and experimental renal diseases characterized by pathologic foot process remodeling, prompting the hypothesis that phosphonephrin signaling directly influences podocyte morphology. To explore this possibility, we generated and analyzed knockin mice with mutations that disrupt nephrin tyrosine phosphorylation and Nck1/2 binding (nephrinY3F/Y3F mice). Homozygous nephrinY3F/Y3F mice developed progressive proteinuria accompanied by structural changes in the filtration barrier, including podocyte foot process effacement, irregular thickening of the glomerular basement membrane, and dilated capillary loops, with a similar but later onset phenotype in heterozygous animals. Furthermore, compared with wild-type mice, nephrinY3F/Y3F mice displayed delayed recovery in podocyte injurymodels. Profilingof nephrin tyrosinephosphorylationdynamics in wild-typemice subjectedtopodocyte injury indicated site-specific differences in phosphorylation at baseline, injury, and recovery, which correlated with loss of nephrin-Nck1/2 association during foot process effacement. Our results define an essential requirement for nephrin tyrosine phosphorylation in stabilizing podocyte morphology and suggest a model in whichdynamic changes in phosphotyrosine-based signaling confer plasticity to thepodocyte actin cytoskeleton.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 2422-2435 |
| Number of pages | 14 |
| Journal | Journal of the American Society of Nephrology |
| Volume | 27 |
| Issue number | 8 |
| DOIs | |
| State | Published - 2016 |
Funding
This work was supported by grants fromthe Kidney Foundation of Canada (to N.J.), Canadian Institutes of Health Research (to N.J.), and the Sick Kids Foundation (to N.J.). L.A.N. and C.E.M. were supported by Natural Sciences and Engineering Research Council of Canada Alexander Graham Bell Canada Graduate Doctoral Scholarships. C.E.M., M.J.P., and A.K.C. were supported by Ontario Graduate Scholarships. N.J. holds a Tier II Canada Research Chair in Eukaryotic Cellular Signaling and was the recipient of a New Investigator Award from the Kidney Research Scientist Core Education and National Training Program.
ASJC Scopus subject areas
- General Medicine
