2003年2月
Cell density and growth-dependent down-regulation of both intracellular calcium responses to agonist stimuli and expression of smooth-surfaced endoplasmic reticulum in MC3T3-E1 osteoblast-like cells
JOURNAL OF BIOLOGICAL CHEMISTRY
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- 巻
- 278
- 号
- 8
- 開始ページ
- 6433
- 終了ページ
- 6439
- 記述言語
- 英語
- 掲載種別
- 研究論文(学術雑誌)
- DOI
- 10.1074/jbc.M210243200
- 出版者・発行元
- AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC
A two-dimensional intracellular Ca2+ ([Ca2+](i)) imaging system was used to examine the relationship between [Ca2+](i) handling and the proliferation of MC3T3-E1 osteoblast-like cells. The resting [Ca2+](i) level in densely cultured cells was 1.5 times higher than the [Ca2+](i) level in sparsely cultured cells or in other cell types (mouse fibroblasts, rat vascular smooth muscle cells, and bovine endothelial cells). A high resting [Ca2+](i) level may be specific for MC3T3-E1 cells. MC3T3-E1 cells were stimulated with ATP (10 mum), caffeine (10 mm), thapsigargin (1 mum), or ionomycin (10 mum), and the effect on the [Ca2+](i) level of MC3T3-E1 cells was studied. The percentage of responding cells and the degree of [Ca2+](i) elevation were high in the sparsely cultured cells and low in densely cultured cells. The rank order for the percentage of responding cells and magnitude of the Ca2+ response to the stimuli was ionomycin > thapsigargin = ATP > caffeine and suggests the existence of differences among the various [Ca2+](i) channels. All Ca2+ responses in the sparsely cultured MC3T3-E1 cells, unlike in other cell types, disappeared after the cells reached confluence. Heptanol treatment of densely cultured cells restored the Ca2+ response, suggesting that cell-cell contact is involved with the confluence-dependent disappearance of the Ca2+ response. Immunohistological analysis of type 1 inositol trisphosphate receptors and electron microscopy showed distinct expression of inositol trisphosphate receptor proteins and smooth-surfaced endoplasmic reticulum in sparsely cultured cells but reduced levels in densely cultured cells. These results indicate that the underlying basis of confluence-dependent [Ca2+](i) regulation is down-regulation of smooth-surfaced endoplasmic reticulum by cell-cell contacts.
- リンク情報
- ID情報
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- DOI : 10.1074/jbc.M210243200
- ISSN : 0021-9258
- Web of Science ID : WOS:000181129400117