Podocytic PKC-alpha is regulated in murine and human diabetes and mediates nephrin endocytosis.
BACKGROUND
Microalbuminuria is an early lesion during the development of diabetic nephropathy. The loss of high molecular weight proteins in the urine is usually associated with decreased expression of slit diaphragm proteins. Nephrin, is the major component of the glomerular slit diaphragm and loss of nephrin has been well described in rodent models of experimental diabetes as well as in human diabetic nephropathy.


RESULTS
In this manuscript we analyzed the role of PKC-alpha (PKCalpha) on endocytosis of nephrin in podocytes. We found that treatment of diabetic mice with a PKCalpha-inhibitor (GO6976) leads to preserved nephrin expression and reduced proteinuria. In vitro, we found that high glucose stimulation would induce PKCalpha protein expression in murine and human podocytes. We can demonstrate that PKCalpha mediates nephrin endocytosis in podocytes and that overexpression of PKCalpha leads to an augmented endocytosis response. After PKC-activation, we demonstrate an inducible association of PKCalpha, PICK1 and nephrin in podocytes. Moreover, we can demonstrate a strong induction of PKCalpha in podocytes of patients with diabetic nephropathy.


CONCLUSIONS
We therefore conclude that activation of PKCalpha is a pathomechanistic key event during the development of diabetic nephropathy. PKCalpha is involved in reduction of nephrin surface expression and therefore PKCalpha inhibition might be a novel target molecule for anti-proteinuric therapy.
Tossidou
Irini
I
Division of Nephrology, Department of Medicine, Hannover Medical School, Hannover, Germany.
Teng
Beina
B
Menne
Jan
J
Shushakova
Nelli
N
Park
Joon-Keun
JK
Becker
Jan U
JU
Modde
Friedrich
F
Leitges
Michael
M
Haller
Hermann
H
Schiffer
Mario
M
D000818:Animals; D003920:Diabetes Mellitus / Q000201:enzymology* / Q000378:metabolism; D003928:Diabetic Nephropathies / Q000209:etiology / Q000473:pathology; D004705:Endocytosis; D015971:Gene Expression Regulation, Enzymologic; D005947:Glucose / Q000494:pharmacology; D006801:Humans; D008565:Membrane Proteins / Q000378:metabolism*; D051379:Mice; D050199:Podocytes / Q000378:metabolism* / Q000502:physiology; D051571:Protein Kinase C-alpha / Q000235:genetics / Q000378:metabolism*
D008565:Membrane Proteins; C113267:nephrin; D051571:Protein Kinase C-alpha; D005947:Glucose