AUTHOR=Wen Jiliang , Chen Zhenghao , Zhao Mengmeng , Zu Shulu , Zhao Shengtian , Wang Shaoyong , Zhang Xiulin TITLE=Cell Deformation at the Air-Liquid Interface Evokes Intracellular Ca2+ Increase and ATP Release in Cultured Rat Urothelial Cells JOURNAL=Frontiers in Physiology VOLUME=12 YEAR=2021 URL=https://www.frontiersin.org/journals/physiology/articles/10.3389/fphys.2021.631022 DOI=10.3389/fphys.2021.631022 ISSN=1664-042X ABSTRACT=

Urothelial cells have been implicated in bladder mechanosensory transduction, and thus, initiation of the micturition reflex. Cell deformation caused by tension forces at an air-liquid interface (ALI) can induce an increase in intracellular Ca²⁺ concentration ([Ca²⁺]_i) and ATP release in some epithelial cells. In this study, we aimed to examine the cellular mechanisms underlying ALI-induced [Ca²⁺]_i increase in cultured urothelial cells. The ALI was created by stopping the influx of the perfusion but maintaining efflux. The [Ca²⁺]_i increase was measured using the Ca²⁺ imaging method. The ALI evoked a reversible [Ca²⁺]_i increase and ATP release in urothelial cells, which was almost abolished by GdCl₃. The specific antagonist of the transient receptor potential vanilloid (TRPV4) channel (HC0674) and the antagonist of the pannexin 1 channel (¹⁰panx) both diminished the [Ca²⁺]_i increase. The blocker of Ca²⁺-ATPase pumps on the endoplasmic reticulum (thapsigargin), the IP3 receptor antagonist (Xest-C), and the ryanodine receptor antagonist (ryanodine) all attenuated the [Ca²⁺]_i increase. Degrading extracellular ATP with apyrase or blocking ATP receptors (P2X or P2Y) with pyridoxalphosphate-6-azophenyl-2',4'-disulfonic acid (PPADS) significantly attenuated the [Ca²⁺]_i increase. Our results suggest that both Ca²⁺ influx via TRPV4 or pannexin 1 and Ca²⁺ release from intracellular Ca²⁺ stores via IP3 or ryanodine receptors contribute to the mechanical responses of urothelial cells. The release of ATP further enhances the [Ca²⁺]_i increase by activating P2X and P2Y receptors via autocrine or paracrine mechanisms.