Abstract 401: Prostaglandin E2 Stimulates Renin Synthesis in Mouse Collecting Duct M-1 Cells via Ep1 Receptor Through Pkc/camp/creb Pathway

Abstract

Prostaglandin E2 (PGE2) plays a major role in regulating renin expression and release by the renal juxtaglomerular (JG) cells. Recently it has been demonstrated that PGE2-dependent upregulation of renin in JG cells is mediated by activation of E prostaoind receptor type 4 (EP4) via cAMP accumulation. Renin is also produced by the principal cells of the collecting ducts (CD) and is upregulated during angiotensin II-dependent hypertension. However, the effects of PGE2 on CD renin remain unknown. Four types of receptors have been described in rat and mouse CD, EP1, EP3 and EP4. Here, we tested the hypothesis that renin is upregulated by PGE2 via activation of EP receptors in mouse CD M-1 cells. By immunostaining we confirmed the presence of EP1, EP3 and EP4 receptors, while EP2 was not detected. A dose response treatment with PGE2 showed increased levels of renin mRNA and protein with a maximum response at 1 μmol/L (mRNA: 19.3 ± 3.0; P<0.05; protein: 3.01 ± 0.08 fold change; P<0.05). To assess which EP receptor is involved in the renin upregulation we used specific EP receptor antagonists: ONO-8711 (EP1; 10 nmol/L), L-798106 (EP3; 10 μmol/L) and AH 23848 (EP4; 10 μmol/L). EP1 antagonist suppressed the PGE2-mediated upregulation of collecting duct renin mRNA and protein (mRNA: 1.0 ± 0.2; protein: 0.98 ± 0.13 fold change; P=NS), while EP4 antagonist only partially decreased it (mRNA: 11.2 ± 2.8; P<0.05; protein: 2.81 ± 0.07 fold change; P<0.05). EP3 antagonist exacerbated the PGE2 mediated-upregulation of renin (mRNA: 50.3 ± 6.0; P<0.05; protein: 3.56 ± 0.08; fold change; P<0.05). Because EP1 is a Gq linked receptor that activates PKC, we further assessed the effects of PKC inhibition using calphostin C and a PKCα dominant negative (DN) on renin expression. Calphostin C and PKCα-DN blunted the PGE2-induced renin upregulation. Importantly, the increases in cAMP levels and phosphorylation of the cAMP response element-binding transcription factor (CREB) mediated by PGE2 were also prevented by both treatments. The results indicate that in mouse CD cells, EP1 receptor activation upregulates renin synthesis via PKC/cAMP/CREB, suggesting that the presence of PGE2 in renal medullary tissues may contribute to the stimulation of collecting duct renin.