Podocyte changes upon induction of albuminuria in Thy-1.1 transgenic mice.
BACKGROUND
Thy-1.1 transgenic mice, characterized by ectopic expression of the Thy-1.1 protein on podocytes, spontaneously develop proteinuria and focal glomerulosclerosis (FGS). Injection of a monoclonal antibody (mAb) directed against the Thy-1.1 protein in young transgenic mice induces a massive albuminuria that is followed by an accelerated FGS within 3 weeks. This albuminuria is complement and leukocyte independent. The time course of proteinuria, the pathogenesis of the acute proteinuria and the dose dependency of FGS are unknown.


METHODS
Albuminuria was measured in Thy-1.1 transgenic mice after injection of different doses of anti-Thy-1.1 mAb and at different time points within the first 24 h after injection. Podocytic foot processes and slit pore diameter were quantitated by electron microscopy. Changes in expression of slit pore constituents (podocin, CD2AP, nephrin and ZO-1), cytoskeleton-associated proteins (actin, alpha-actinin, ezrin and synaptopodin), the GDH-podocyte adhesion molecules alpha(3)-integrin, and heparan sulfate were studied by immunofluorescence. FGS was scored by light microscopy at 3 weeks after induction of albuminuria.


RESULTS
Albuminuria in Thy-1.1 transgenic mice was observed within 10 min after anti-Thy-1.1 mAb injection. This rapid development of albuminuria was accompanied by a reduction in number of podocytic foot processes from 20.0 +/- 0.7/10 microm glomerular basement membrane (GBM) in saline-treated transgenic mice to 8.0 +/- 0.5 and 2.2 +/- 0.2 in anti-Thy-1.1-treated mice, at 10 min and 8 h after treatment, respectively. In addition, we observed a significant decrease in width of remaining slit pores, from 32.7 +/- 1.1 to 26.8 +/- 1.4 nm at 10 min after mAb injection. By immunofluorescence, we did not observe major changes in the expression pattern of any of the proteins studied. There was no correlation between the injected dose of the anti-Thy-1.1 mAb and the acute albuminuria. In contrast, the percentage of FGS at 3 weeks correlated with the dose, and a significant correlation between the percentage of FGS and the time-averaged albuminuria over the 3 week study period (P < 0.001) was found.


CONCLUSIONS
Injection of mAb directed against the Thy-1.1 protein, in young non-albuminuric Thy-1.1 transgenic mice, induced an acute albuminuria within 10 min, which was accompanied by foot process effacement. Notably, we observed a decrease in slit pore width although the expression of slit pore proteins was unchanged. Also, the acute albuminuria could not be related to alterations in cytoskeleton-associated proteins, the GBM adhesion molecule alpha(3)-integrin or heparan sulfate in the GBM. The dose-dependent development of FGS and the correlation between the percentage FGS and time-averaged albuminuria suggest that, in our model, FGS is a consequence of podocyte injury. However, the data leave open the possibility that albuminuria itself contributes to FGS development. The Thy-1.1 transgenic mouse model is an excellent model to study further the relationship between podocytic injury, albuminuria and the development of FGS.
Smeets
Bart
B
Department of Pathology, University Medical Center Nijmegen, PO Box 9101, 6500 HB Nijmegen, The Netherlands. b.smeets@pathol.umcn.nl
Dijkman
Henry B P M
HB
te Loeke
Nathalie A J M
NA
van Son
Jacco P H F
JP
Steenbergen
Eric J
EJ
Assmann
Karel J M
KJ
Wetzels
Jack F M
JF
Groenen
Patricia J T A
PJ
D000419:Albuminuria / Q000276:immunology* / Q000503:physiopathology; D000818:Animals; D000911:Antibodies, Monoclonal / Q000009:adverse effects; D004305:Dose-Response Relationship, Drug; D004847:Epithelial Cells / Q000276:immunology*; D005923:Glomerulosclerosis, Focal Segmental / Q000150:complications / Q000276:immunology* / Q000503:physiopathology; D007678:Kidney Glomerulus / Q000276:immunology* / Q000503:physiopathology; D051379:Mice; D008822:Mice, Transgenic / Q000276:immunology; D023421:Models, Animal; D018800:Thy-1 Antigens / Q000276:immunology*; D013997:Time Factors
D000911:Antibodies, Monoclonal; D018800:Thy-1 Antigens