Real-time monitoring of cellular signaling in salivary secretion by multiphoton excitation imaging
| Project/Area Number |
15591975
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Functional basic dentistry
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| Research Institution | Health Sciences University of Hokkaido, School of Dentistry |
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Principal Investigator |
TOJYO Yosuke Health Sciences University of Hokkaido, School of Dentistry, Professor, 歯学部, 教授 (90111731)
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| Co-Investigator(Kenkyū-buntansha) |
TANIMURA Akihiko Health Sciences University of Hokkaido, School of Dentistry, Associate Professor, 歯学部, 助教授 (70217149)
NEZU Akihiro Health Sciences University of Hokkaido, School of Dentistry, Instructor, 歯学部, 助手 (00305913)
MORITA Takao Health Sciences University of Hokkaido, School of Dentistry, Instructor, 歯学部, 助手 (20326549)
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| Project Period (FY) |
2003 – 2004
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| Project Status |
Completed (Fiscal Year 2004)
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| Budget Amount *help |
¥3,600,000 (Direct Cost: ¥3,600,000)
Fiscal Year 2004: ¥2,000,000 (Direct Cost: ¥2,000,000)
Fiscal Year 2003: ¥1,600,000 (Direct Cost: ¥1,600,000)
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| Keywords | multiphoton laser microscopy / salivary gland / calcium signal / calcium wave / duct cells / acinar cells / tissue slices / fura-2 / 細胞内カルシウムイオン / エピネフリン / 多光子励起法 / レーザー顕微鏡 / イノシトール1,4,5-三リン酸 / カルシウムストア / 耳下腺細胞 |
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| Research Abstract |
1.The agonist-induced Ca^<2+> signaling in rat salivary slices and cell aggregates was visualized using multiphoton excitation microscopy. The Ca^<2+> images were compared with the results obtained by a conventional C^<2+> imaging system (ARGUS HiSCA).
2. Salivary cell aggregates were loaded with fura-2 and the changes in fluorescence were monitored. Stimulation with a low concentration (0.1μM) of epinephrine (Epi) induced an increase in in a part of duct cells, while most of the cells responded to its higher concentration (1μM). The Ca^<2+> response was initiated in a duct cell and followed by a rise in [Ca^<2+>]i in the neighboring cells.
3. We characterized the duct cells based on their [Ca^<2+>]i-responses to Epi and ATP. About 40 % of duct cells were highly responsive to Epi and ATP. Another 40 % of them were less responsive to Epi but highly responsive to ATP. About 2 % of the cells responded only to ATP, and 10 % of the cells responded to neither Epi nor ATP.
4. Stimulation with 1μM Epi induced an intracellular propagating-[Ca^<2+>]i wave from luminal to basal area of duct cells.
5. The [Ca^<2+>]i responses in rat parotid gland slices were monitored by two-photon excitation microscopy. Stimulation with 1μEpi caused simultaneous rises in [Ca^<2+>]i in all acinar cells, while the initiation time of [Ca^<2+>]i responses varied with duct cells. The intercellular Cat wave propagation occurred in some duct cells.
6. The agonist induced [Ca^<2+>]i responses were compared in rat submandibular acini and ducts. Acini were highly responsive to carbachol and substance P, while duct; were highly responsive to Epi but did not respond to substance P.
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Report
(3 results)
Research Products
(3 results)
