Parathyroid hormone (PTH) regulates the sodium chloride cotransporter via Ras guanyl releasing protein 1 (Ras-GRP1) and extracellular signal-regulated kinase (ERK)1/2 mitogen-activated protein kinase (MAPK) pathway

The sodium chloride cotransporter (NCC) is the principal salt absorptive pathway in the mammalian distal convoluted tubule (DCT) and is the site of action of thiazide diuretics. Using a mammalian cell model system to assess NCC function, we demonstrated previously that Ras guanyl releasing protein 1 (Ras-GRP1) mediates phorbol ester-induced suppression of the function and surface expression of NCC in a protein kinase C (PKC)-independent and extracellular signal-regulated kinase (ERK)1/2-dependent manner. Given that phorbol esters are functional analogs of diacylglycerol (DAG), this finding suggested a potential physiologic regulation of NCC by DAG. The parathyroid hormone (PTH) receptor is a G-protein-coupled receptor that is expressed in the DCT and activates PLC resulting in the generation of DAG. In this article, we demonstrate that PTH suppresses NCC function via a PLC/Ras-GRP1/ERK pathway. A functional assessment of NCC measuring thiazide-sensitive 22Na+ flux revealed that PTH suppresses NCC function. The inhibition of PLC prevented the suppression of NCC, indicating that PLC was necessary for this effect. Inhibitors of PKC and protein kinase A (PKA) had no effect on this suppression, but mitogen-activated protein kinase (MAPK) inhibitors prevented the PTH effect completely. Ras-GRP1 activates the MAPK pathway though activation of the small G-protein Ras. Gene silencing of Ras-GRP1 prevented the PTH-mediated suppression of NCC activity, the activation of the H-Ras isoform of Ras, and the activation of ERK1/2 MAPK. This finding confirmed the critical role of Ras-GRP1 in mediating the PTH-induced suppression of NCC activity through stimulation of the MAPK pathway.