CEDIE   05498
CENTRO DE INVESTIGACIONES ENDOCRINOLOGICAS "DR. CESAR BERGADA"
Unidad Ejecutora - UE
artículos
Título:
Signaling cascade of insulin-induced stimulation of L-dopa uptake in renal proximal tubule cells
Autor/es:
CARRANZA ANDREA; MUSOLINO PATRICIA; VILLAR MARCELO; NOWICKI SUSANA
Revista:
AMERICAN JOURNAL OF PHYSIOLOGY-CELL PHYSIOLOGY
Editorial:
The American Physiology Society
Referencias:
Lugar: Bethesda; Año: 2008 vol. 295 p. 1602 - 1609
ISSN:
0363-6143
Resumen:
The inward L-dopa transport supplies renal proximal tubule cells (PTCs) with the precursor for dopamine synthesis. We have previously described insulin-induced stimulation of L-dopa uptake into PTCs. In the present paper we examined insulin- related signaling pathways involved in the increase of L-dopa transport into isolated rat PTCs.  Insulin (50-500 mU/ml) increased L-dopa uptake by PTCs reaching the maximal increment (60% over the control) at 200 mU/ml. At this concentration insulin also increased insulin receptor tyrosine phosphorylation. Both effects were abrogated by the tyrosine kinase inhibitor genistein (5 mM). In line, inhibition of the protein tyrosine phosphatase by pervanadate (0.2-100 mM) caused a concentration-dependent increase in both, the uptake of L-dopa (up to 400%), and protein tyrosine phosphorylation. A synergistic effect between pervanadate and insulin on L-dopa uptake was observed only when threshold (0.2 mM), but not maximal (5 mM), concentrations of pervanadate were assayed. Insulin- induced stimulation of L-dopa uptake was also abolished by inhibition of phosphatydil inositol 3-kinase (PI3K, wortmanine 100nM, and LY294002 25 mM) and protein kinase C (PKC, RO81220, 1 mM). Insulin-induced activation of PKC-z was confirmed in vitro by its translocation from the cytosol to the membrane fraction, and in vivo by immunohistochemistry studies. Insulin caused a wortmanine-sensitive increase in Akt/protein kinase B (Akt/PKB) phosphorylation and a dose-dependent translocation of Akt/PKB to the membrane fraction. Our findings suggest that insulin activates PKC-z, and Akt/PKB downstream of PI3K, and these pathways contribute to the insulin-induced increase of L-dopa uptake into PTCs.