Intracellular cAMP signaling-induced Ca 2+ influx mediated by calcium homeostasis modulator 1 (CALHM1) in human odontoblasts.

Autor: Kimura M; Department of Physiology, Tokyo Dental College, Tokyo, 101-0061, Japan., Nomura S; Department of Physiology, Tokyo Dental College, Tokyo, 101-0061, Japan., Ouchi T; Department of Physiology, Tokyo Dental College, Tokyo, 101-0061, Japan., Kurashima R; Department of Physiology, Tokyo Dental College, Tokyo, 101-0061, Japan., Nakano R; Laboratory for Mucosal Immunity, Center for Integrative Medical Sciences, RIKEN Yokohama Institute, Yokohama, 230-0045, Japan.; Japan Animal Specialty Medical Institute (JASMINE), Yokohama, 224-0001, Japan., Sekiya H; Department of Endodontics, Tokyo Dental College, Tokyo, 101-0061, Japan., Kuroda H; Department of Physiology, Tokyo Dental College, Tokyo, 101-0061, Japan.; Department of Dental Anesthesiology, Kanagawa Dental University, Yokosuka, 238-8570, Japan., Kono K; Department of Physiology, Tokyo Dental College, Tokyo, 101-0061, Japan., Shibukawa Y; Department of Physiology, Tokyo Dental College, Tokyo, 101-0061, Japan. yshibuka@tdc.ac.jp.
Jazyk: angličtina
Zdroj: Pflugers Archiv : European journal of physiology [Pflugers Arch] 2024 Nov 12. Date of Electronic Publication: 2024 Nov 12.
DOI: 10.1007/s00424-024-03038-4
Abstrakt: In odontoblasts, intracellular Ca 2+ signaling plays key roles in reactionary dentin formation and generation of dentinal pain. Odontoblasts also express several G s protein-coupled receptors that promote production of cyclic AMP (cAMP). However, the crosstalk between intracellular cAMP and Ca 2+ signaling, as well as the role of cAMP in the cellular functions of odontoblasts, remains unclear. In this study, we measured intracellular cAMP levels and intracellular free Ca 2+ concentration ([Ca 2+ ] i ). We also investigated the effect of intracellular cAMP on mineralization by the odontoblasts. In the presence of extracellular Ca 2+ , the application of forskolin (adenylyl cyclase activator) or isoproterenol (G s protein-coupled beta-2 adrenergic receptor agonist) increased intracellular cAMP levels and [Ca 2+ ] i in odontoblasts. The [Ca 2+ ] i increases could not be observed by removing extracellular Ca 2+ , indicating that cAMP is capable to activate Ca 2+ entry. Forskolin-induced [Ca 2+ ] i increase was inhibited by a protein kinase A inhibitor in odontoblasts. The [Ca 2+ ] i increase was sensitive to Gd 3+ , 2APB, or Zn 2+ but not verapamil, ML218, or La 3+ . In immunofluorescence analyses, odontoblasts were immunopositive for calcium homeostasis modulator 1 (CALHM1), which was found close to ionotropic ATP receptor subtype, P2X 3 receptors. When CALHM1 was knocked down, forskolin-induced [Ca 2+ ] i increase was suppressed. Alizarin red and von Kossa staining showed that forskolin decreased mineralization. These findings suggest that activation of adenylyl cyclase elicited increases in the intracellular cAMP level and Ca 2+ influx via protein kinase A activation in odontoblasts. Subsequent cAMP-dependent Ca 2+ influx was mediated by CALHM1 in odontoblasts. In addition, the intracellular cAMP signaling pathway in odontoblasts negatively mediated dentinogenesis.
Competing Interests: Declarations Ethical approval Not applicable. Competing interests The authors declare no competing interests.
(© 2024. The Author(s).)
Databáze: MEDLINE
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