ANG II constricts descending vasa recta (DVR) through Ca²⁺ signaling in pericytes. We examined the role of PKC DVR pericytes isolated from the rat renal outer medulla. The PKC blocker staurosporine (10 μM) eliminated ANG II (10 nM)-induced vasoconstriction, inhibited pericyte cytoplasmic Ca²⁺ concentration ([Ca²⁺]_cyt) elevation, and blocked Mn²⁺ influx into the cytoplasm. Activation of PKC by either 1,2-dioctanoyl-sn-glycerol (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 ANG II- and PDBu-induced PKC stimulated DVR generation of reactive oxygen species (ROS), measured by oxidation of dihydroethidium (DHE). The effect of ANG II was only significant when ANG II AT₂ receptors were blocked with PD-123319 (10 nM). PDBu augmentation of DHE oxidation was blocked by either TEMPOL (1 mM) or diphenylene iodonium (10 μM). We conclude that ANG II and PKC activation increases DVR pericyte [Ca²⁺]_cyt, divalent ion conductance into the cytoplasm, and ROS generation.