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1 Department of Medicine, Division of Nephrology, University of Maryland, Baltimore, MD, USA
2 Department of Medicine, Division of Nephrology, University of Maryland, Baltimore, MD, USA; Department of Physiology, University of Maryland, Baltimore, MD, USA
* To whom correspondence should be addressed. E-mail: tpallone{at}medicine.umaryland.edu.
Angiotensin II (AngII) constricts descending vasa recta (DVR) through Ca2+ signaling in pericytes. We examined the role of protein kinase C (PKC) DVR pericytes isolated from the rat renal outer medulla. The PKC blocker, staurosporine (10 µM), eliminated AngII (10 nM) induced vasoconstriction, inhibited pericyte cytoplasmic Ca2+ ([Ca2+]CYT) elevation and blocked Mn2+ influx into the cytoplasm. Activation of PKC by either 1, 2-dioctanoyl-sn-glycerol (DiOG, 10 µM) or phorbol 12, 13-dibutyrate (PDBu, 1 µM) induced both vasoconstriction and pericyte [Ca2+]CYT elevation. Diltiazem (10 µM) blocked the ability of PDBu to increase pericyte [Ca2+]CYT and enhance Mn2+ influx. Both AngII and PDBu induced PKC stimulated DVR generation of reactive oxygen species (ROS), measured by oxidation of dihydroethidium (DHE). The effect of AngII was only signficant when AngII, AT2 receptors were blocked with PD123319 (10 nM). PDBu augmentation of DHE oxidation was blocked by either tempol (1 mM) or diphenylene iodonium (10 µM). We conclude that AngII and PKC activation increases DVR pericyte [Ca2+]CYT, divalent ion conductance into the cytoplasm and ROS generation.
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