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Am J Physiol Renal Physiol (April 4, 2006). doi:10.1152/ajprenal.00448.2005
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Submitted on November 9, 2005
Accepted on March 29, 2006

Autocrine VEGF system in podocytes regulates podocin and its interaction with CD2AP

Fangxia Guan1, Guillermo Villegas1, Jason Teichman2, Peter Mundel3, and Alda Tufro2*

1 Pediatrics/Nephrology, Albert Einstein College of Medicine, Bronx, New York, United States; Developmental and Molecular Biology, Albert Einstein College of Medicine, New York
2 Pediatrics/Nephrology, Albert Einstein College of Medicine, Bronx, New York, United States; Developmental and Molecular Biology, Albert Einstein College of Medicine
3 Medicine, Mount Sinai School of Medicine, New York, New York, United States

* To whom correspondence should be addressed. E-mail: atufro{at}aecom.yu.edu.

Vascular endothelial growth factor (VEGF-A) signaling is required for endothelial cell differentiation, vasculogenesis, angiogenesis and vascular patterning. During kidney morphogenesis, podocyte VEGF-A guides endothelial cells towards developing glomeruli. Podocyte VEGF-A expression continues throughout life but its function after completion of development remains unclear. Here we examined the expression of VEGF-A and its receptors VEGFR1, VEGFR2, NP1 and NP2 in conditionally immortalized mouse podocytes cultured in undifferentiated and differentiated conditions using RT-PCR and western analysis. VEGF-A secretion was assessed by ELISA and western analysis. Upon podocyte differentiation VEGF-A protein expression and secretion increased three- fold. Differentiated podocytes expressed eight-fold higher VEGFR2 mRNA levels than undifferentiated podocytes, whereas VEGFR1, sVEGFR1, NP1 and NP2 mRNA levels were similar. We examined the regulation and function of the VEGF-A system by exposing differentiated podocytes to recombinant VEGF165 [20ng/ml] or control media for 24h. VEGF165 induced a two-fold increase in VEGFR2 mRNA and protein levels whereas VEGFR1, sVEGFR1, NP1 and NP2 mRNA levels remained unchanged. VEGF165 induced VEGFR2 phosphorylation. VEGF165 reduced podocyte apoptosis ~40% whereas anti-VEGFR2 neutralizing antibody enhanced it two-fold. We determined that VEGF-A signaling regulates slit diaphragm proteins by inducing a dose-response podocin upregulation and increasing its interaction with CD2AP. The data indicate that podocytes in culture have a functional autocrine VEGF-A system that is regulated by differentiation and ligand availability. VEGF-A functions in podocytes include promoting survival through VEGFR2, inducing podocin upregulation and increasing podocin/CD2AP interaction.




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