Parathyroid hormone controls paracellular Ca ²⁺ transport in the thick ascending limb by regulating the tight-junction protein Claudin14

Abstract

Renal Ca²⁺ reabsorption is essential for maintaining systemic Ca²⁺ homeostasis and is tightly regulated through the parathyroid hormone (PTH)/PTHrP receptor (PTH1R) signaling pathway. We investigated the role of PTH1R in the kidney by generating a mouse model with targeted deletion of PTH1R in the thick ascending limb of Henle (TAL) and in distal convoluted tubules (DCTs): <i>Ksp-cre;Pth1r</i>^<i>fl/fl</i> Mutant mice exhibited hypercalciuria and had lower serum calcium and markedly increased serum PTH levels. Unexpectedly, proteins involved in transcellular Ca²⁺ reabsorption in DCTs were not decreased. However, claudin14 (Cldn14), an inhibitory factor of the paracellular Ca²⁺ transport in the TAL, was significantly increased. Analyses by flow cytometry as well as the use of <i>Cldn14-lacZ</i> knock-in reporter mice confirmed increased Cldn14 expression and promoter activity in the TAL of <i>Ksp-cre;Pth1r</i>^<i>fl/fl</i> mice. Moreover, PTH treatment of HEK293 cells stably transfected with CLDN14-GFP, together with PTH1R, induced cytosolic translocation of CLDN14 from the tight junction. Furthermore, mice with high serum PTH levels, regardless of high or low serum calcium, demonstrated that PTH/PTH1R signaling exerts a suppressive effect on Cldn14. We therefore conclude that PTH1R signaling directly and indirectly regulates the paracellular Ca²⁺ transport pathway by modulating Cldn14 expression in the TAL. Finally, systemic deletion of Cldn14 completely rescued the hypercalciuric and lower serum calcium phenotype in <i>Ksp-cre;Pth1r</i>^<i>fl/fl</i> mice, emphasizing the importance of PTH in inhibiting Cldn14. Consequently, suppressing CLDN14 could provide a potential treatment to correct urinary Ca²⁺ loss, particularly in patients with hypoparathyroidism.