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Real-time monitoring of cellular signaling in salivary secretion by multiphoton excitation imaging

通过多光子激发成像实时监测唾液分泌中的细胞信号传导

基本信息

批准号：
15591975
负责人：
TOJYO Yosuke
金额：
$ 2.3万
依托单位：
Health Sciences University of Hokkaido, School of Dentistry
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2003
资助国家：
日本
起止时间：
2003 至 2004
项目状态：
已结题

项目摘要

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.

1.用多光子激发显微镜观察激动剂诱导的大鼠唾液切片和细胞聚集体中的Ca^2+信号。将Ca^2+图像与常规C^2+成像系统（ARGUS HiSCA）获得的结果进行比较。用fura-2加载唾液细胞聚集体，并监测荧光的变化。低浓度（0.1μM）的肾上腺素（Epi）刺激可引起部分导管细胞的Epi升高，而高浓度（1μM）的Epi刺激可引起大部分导管细胞的Epi升高。导管细胞的Ca^<2+>反应开始，随后邻近细胞的[Ca^<2+]i升高.我们基于导管细胞对Epi和ATP的[Ca^<2+>] i反应来表征导管细胞。约40%的导管细胞对Epi和ATP高度反应.另外40%的人对Epi反应较低，但对ATP反应较高。大约2%的细胞只对ATP有反应，10%的细胞对Epi和ATP都没有反应。用1μM Epi刺激诱导导管细胞的胞内传播-[Ca^<2+>]i波从管腔到基底区。用双光子激发显微镜观察大鼠腮腺切片[Ca^<2+>]i的变化。1μEpi刺激引起所有腺泡细胞内[Ca^<2+>]i同时升高，而[Ca^<2+>]i反应的起始时间因导管细胞而异。细胞间Cat波在部分导管细胞中传播.在大鼠下颌下腺腺泡和导管中比较激动剂诱导的[Ca^<2+>]i反应。腺泡对氨甲酰胆碱和P物质有高度反应，而导管对Epi有高度反应，但对P物质无反应。