Plasma Membrane Ca 2+ -ATPase in Rat and Human Odontoblasts Mediates Dentin Mineralization.

Autor: Kimura M; Department of Physiology, Tokyo Dental College, 2-9-18, Kanda-Misaki-cho, Chiyoda-ku, Tokyo 101-0061, Japan., Mochizuki H; Department of Physiology, Tokyo Dental College, 2-9-18, Kanda-Misaki-cho, Chiyoda-ku, Tokyo 101-0061, Japan., Satou R; Department of Epidemiology and Public Health, Tokyo Dental College, Chiyodaku, Tokyo 101-0061, Japan., Iwasaki M; Department of Epidemiology and Public Health, Tokyo Dental College, Chiyodaku, Tokyo 101-0061, Japan., Kokubu E; Department of Microbiology, Tokyo Dental College, Chiyodaku, Tokyo 101-0061, Japan., Kono K; Department of Physiology, Tokyo Dental College, 2-9-18, Kanda-Misaki-cho, Chiyoda-ku, Tokyo 101-0061, Japan., Nomura S; Department of Physiology, Tokyo Dental College, 2-9-18, Kanda-Misaki-cho, Chiyoda-ku, Tokyo 101-0061, Japan., Sakurai T; Department of Physiology, Tokyo Dental College, 2-9-18, Kanda-Misaki-cho, Chiyoda-ku, Tokyo 101-0061, Japan., Kuroda H; Department of Physiology, Tokyo Dental College, 2-9-18, Kanda-Misaki-cho, Chiyoda-ku, Tokyo 101-0061, Japan.; Department of Dental Anesthesiology, Kanagawa Dental University, 1-23, Ogawacho, Kanagawa, Yokosuka-shi 238-8570, Japan., Shibukawa Y; Department of Physiology, Tokyo Dental College, 2-9-18, Kanda-Misaki-cho, Chiyoda-ku, Tokyo 101-0061, Japan.
Jazyk: angličtina
Zdroj: Biomolecules [Biomolecules] 2021 Jul 10; Vol. 11 (7). Date of Electronic Publication: 2021 Jul 10.
DOI: 10.3390/biom11071010
Abstrakt: Intracellular Ca 2+ signaling engendered by Ca 2+ influx and mobilization in odontoblasts is critical for dentinogenesis induced by multiple stimuli at the dentin surface. Increased Ca 2+ is exported by the Na + -Ca 2+ exchanger (NCX) and plasma membrane Ca 2+ -ATPase (PMCA) to maintain Ca 2+ homeostasis. We previously demonstrated a functional coupling between Ca 2+ extrusion by NCX and its influx through transient receptor potential channels in odontoblasts. Although the presence of PMCA in odontoblasts has been previously described, steady-state levels of mRNA-encoding PMCA subtypes, pharmacological properties, and other cellular functions remain unclear. Thus, we investigated PMCA mRNA levels and their contribution to mineralization under physiological conditions. We also examined the role of PMCA in the Ca 2+ extrusion pathway during hypotonic and alkaline stimulation-induced increases in intracellular free Ca 2+ concentration ([Ca 2+ ] i ). We performed RT-PCR and mineralization assays in human odontoblasts. [Ca 2+ ] i was measured using fura-2 fluorescence measurements in odontoblasts isolated from newborn Wistar rat incisor teeth and human odontoblasts. We detected mRNA encoding PMCA1-4 in human odontoblasts. The application of hypotonic or alkaline solutions transiently increased [Ca 2+ ] i in odontoblasts in both rat and human odontoblasts. The Ca 2+ extrusion efficiency during the hypotonic or alkaline solution-induced [Ca 2+ ] i increase was decreased by PMCA inhibitors in both cell types. Alizarin red and von Kossa staining showed that PMCA inhibition suppressed mineralization. In addition, alkaline stimulation (not hypotonic stimulation) to human odontoblasts upregulated the mRNA levels of dentin matrix protein-1 (DMP-1) and dentin sialophosphoprotein (DSPP). The PMCA inhibitor did not affect DMP-1 or DSPP mRNA levels at pH 7.4-8.8 and under isotonic and hypotonic conditions, respectively. We also observed PMCA1 immunoreactivity using immunofluorescence analysis. These findings indicate that PMCA participates in maintaining [Ca 2+ ] i homeostasis in odontoblasts by Ca 2+ extrusion following [Ca 2+ ] i elevation. In addition, PMCA participates in dentinogenesis by transporting Ca 2+ to the mineralizing front (which is independent of non-collagenous dentin matrix protein secretion) under physiological and pathological conditions following mechanical stimulation by hydrodynamic force inside dentinal tubules, or direct alkaline stimulation by the application of high-pH dental materials.
Databáze: MEDLINE
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