Deprecated: Implicit conversion from float 233.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534
Deprecated: Implicit conversion from float 233.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534
Deprecated: Implicit conversion from float 233.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534
Deprecated: Implicit conversion from float 233.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534
Deprecated: Implicit conversion from float 233.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534
Deprecated: Implicit conversion from float 233.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534
Deprecated: Implicit conversion from float 233.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534
Deprecated: Implicit conversion from float 267.2 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534
Deprecated: Implicit conversion from float 267.2 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534
Warning: imagejpeg(C:\Inetpub\vhosts\kidney.de\httpdocs\phplern\28604827
.jpg): Failed to open stream: No such file or directory in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 117 PLoS+One
2017 ; 12
(6
): e0179217
Nephropedia Template TP
gab.com Text
Twit Text FOAVip
Twit Text #
English Wikipedia
A novel in vivo method to quantify slit diaphragm protein abundance in murine
proteinuric kidney disease
#MMPMID28604827
Haase R
; Potthoff SA
; Meyer-Schwesinger C
; Frosch C
; Wiech T
; Panzer U
; Königshausen E
; Stegbauer J
; Sellin L
; Rump LC
; Quack I
; Woznowski M
PLoS One
2017[]; 12
(6
): e0179217
PMID28604827
show ga
Injury of the glomerular filter causes proteinuria by disrupting the sensitive
interplay of the glomerular protein network. To date, studies of the expression
and trafficking of glomerular proteins have been mostly limited to in vitro or
histologic studies. Here, we report a novel in vivo biotinylation assay that
allows the quantification of surface expression of glomerular proteins in mice.
Kidneys were perfused in situ with biotin before harvest. Afterwards glomeruli
were isolated and lyzed. The protein of interest was separated by
immunoprecipitation and the amount of surface-expressed protein was quantified by
Western blot analysis with streptavidin staining. As proof-of-concept, we
examined the presence of nephrin in the slit diaphragm in two well-established
murine models of proteinuric kidney disease: nephrotoxic nephritis and adriamycin
nephropathy. In proteinuric animals, significantly less nephrin was detected in
the slit diaphragm. When proteinuria decreased once again during the course of
disease, the amount of surface nephrin returned to the baseline. Our present
results suggest that our assay is a valuable tool to study the glomerular filter
in proteinuric kidney diseases. Note that the assay is not limited to proteins
expressed in the slit diaphragm, and all surface proteins that are accessible to
biotin perfusion and immunoprecipitation qualify for this analysis.