Renal Na+ excretion consequent to pharmacogenetic activation of Gq-DREADD in principal cells

Stimulation of metabotropic G(q)-coupled purinergic P2Y(2) receptors decreases activity of the epithelial Na+ channel (ENaC) in renal principal cells of the distal nephron. The physiological consequences of P2Y2 receptor signaling disruption in the P2Y(2) receptor knockout mouse are decreased Na+ excretion and increased arterial blood pressure. However, because of the global nature of this knockout model, the quantitative contribution of ENaC and distal nephron compared with that of upstream renal vascular and tubular elements to changes in urinary excretion and arterial blood pressure is obscure. Moreover, it is uncertain whether stimulation of P2Y(2) receptor inhibition of ENaC is sufficient to drive renal (urinary) Na+ excretion (UNaV). Here, using a pharmacogenetic approach and selective agonism of the P2Y(2) receptor, we test the sufficiency of targeted stimulation of G(q) signaling in principal cells of the distal nephron and P2Y(2) receptors to increase UNaV. Selective stimulation of the P2Y(2) receptor with the ligand MRS2768 decreased ENaC activity in freshly isolated tubules (as assessed by patch-clamp electrophysiology) and increased UNaV (as assessed in metabolic cages). Similarly, selective agonism of hM3Dq-designer receptors exclusively activated by designer drugs (DREADD) restrictively expressed in principal cells of the distal nephron with clozapine-N-oxide decreased ENaC activity and, consequently, increased UNaV. Clozapine-N-oxide, when applied to control littermates, failed to affect ENaC and UNaV. This study represents the first use of pharmacogenetic (DREADD) technology in the renal tubule and demonstrated that selective activation of the P2Y(2) receptor and G(q) signaling in principal cells is sufficient to promote renal salt excretion.