CHARACTERIZATION OF SODIUM-CALCIUM EXCHANGE IN RABBIT RENAL ARTERIOLES

Experiments were performed to test the hypothesis that renal arteriole s exhibit Na-Ca exchange capability and that this process is regulated by protein kinase C (PKC). Glomeruli with attached arterioles were di ssected from rabbit kidney and loaded with fura-2 for measurement of i ntracellular calcium concentration ([Ca2+](i)) using microscope-based photometry. In tissue bathed in Ringer's solution containing 150 mM Na + and 1.5 mM Ca2+, afferent and efferent arteriolar [Ca2+](i) averaged 136 +/- 6 and 154 +/- 7 nM, respectively. Removal of extracellular Na + increased afferent arteriolar [Ca2+](i) by 70 +/- 7 nM, while effere nt arteriolar [Ca2+](i) only increased by 39 +/- 5 nM (P < 0.01 vs. af ferent arteriole). These responses were inhibited by 6 mM Ni2- and req uired extracellular Ca2+, but were unaffected by 10 mu M diltiazem. Af ter incubation in 500 mu M ouabain, 5 mu M monensin, and 5 mu M nigeri cin, [Ca2+](i) responses to removal of extracellular Na+ were exaggera ted significantly, averaging 174 +/- 50 nM in afferent arterioles and 222 +/- 82 nM in efferent arterioles (NS vs. afferent arterioles). Mor eover, responses to removal of extracellular Na+ were enhanced by 100 nM phorbol 12-myristate 13-acetate, an effect which was blocked by PKC inhibition (25 nM K252b). These data indicate that both afferent and efferent arterio]es express the Na-Ca exchanger, and that PKC activity impacts on exchange capacity in these vessels.