The signaling pathway mediating modulation of Na(+)-ATPase of proximal tubule cells by atrial natriuretic peptides (ANP) and urodilatin through receptors located in luminal and basolateral membranes (BLM) is investigated. In isolated BLM, 10(-11)M ANP or 10(-11)M urodilatin inhibited the enzyme activity (50%). Immunodetection revealed the presence of NPR-A in BLM and LLC-PK1 cells. Both compounds increased protein kinase G (PKG) activity (80%) and this effect did not occur with 10(-6)M LY83583, a specific inhibitor of guanylyl cyclase. The inhibitory effect of these peptides on Na(+)-ATPase activity did not occur after addition of 10(-6)M KT5823, a specific inhibitor of PKG. LLC-PK1 cells were used to investigate if ANP and urodilatin change the activity of sodium pumps by luminal receptor interaction. ANP and urodilatin inhibited Na(+)-ATPase activity (50%), with maximal effect at 10(-10)M, similar to 10(-7)M db-cGMP, and did not occur with 10(-7)M LY83583, a guanylyl cyclase inhibitor. ANP and urodilatin specifically inhibit Na(+)-ATPase activity by activation of the cGMP/PKG pathway through NPR-A located in luminal membrane and BLM, increasing understanding of the mechanism of natriuretic peptides on renal sodium excretion, with proximal tubule Na(+)-ATPase one possible target.
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