Study on molecular Mechanism of Calcium Waves Using Rapid Scanning Confocal Microscopy

快速扫描共焦显微镜研究钙波的分子机制

• 批准号: 06670131
• 负责人: KAWANISHI Toru
• 金额: $ 1.34万
• 依托单位: National Institute of Health Sciences
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1994
• 资助国家: 日本
• 起止时间: 1994 至 1995
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-06670131/
• 关键词: confocal laser scanning microscopy; calcium ion; calcium wave; fluorescent probes; liver; hepatocyte

The mechanism of calcium waves was analyzed in primary cultured rat hepatocytes using rapid scanning confocal microscopy. Hormonal stimulations by phenylephrine, vasopressin and ATP and a proliferative stimulation by HGF induced repetitive increases of cytosolic free Ca^<2+> concentration ([Ca^<2+>] i). The increases in [Ca^<2+>] i originated from a specific same origin adjacent to the cell membrane and propagated across the cell like waves. In each cell the locus where the Ca^<2+> waves induced by an agonist wre originated was the same as those where the Ca^<2+> waves induced by the other agonists were originated. There was also a correlation in the peak height in the Ca^<2+> weaves induced by different agonists in each cell. However, there was no correlation between sensitivity of each cell to the agonists, which were measured as latent periods prior to Ca^<2+> rises after an addition of the agonists. The kinetic analysis of Ca^<2+> transients in several small regions in the cells suggested the followings : (1) the spatial patterns of Ca^<2+> waves were not depending on the increases of inostiol 1,4,5 triphosphate (Ins (1,4,5) P_3) in the limited parts of the cells and its propagation, but on the distribution of Ins (1,4,5) P_3-sensitive Ca^<2+> pools ; (2) the sensitivity of each cell to agonists was highly depending on the concentration of Ins (1,4,5) P_3 formed.

用快速扫描共聚焦显微镜分析了原代培养大鼠肝细胞内钙波的发生机制。苯肾上腺素、加压素和三磷酸腺苷的激素刺激以及HGF的增殖刺激引起细胞内游离钙离子浓度([Ca^&lt；2+&gt；]i)反复升高。[Ca^&lt；2+&gt；]i的增加起源于细胞膜附近的同一特定来源，并以波的形式在细胞内传播。在每个细胞中，一种激动剂诱发的Ca^&lt；2+&gt；波的起始点与其他激动剂诱发的Ca^&lt；2+&gt；波的起始点相同。不同激动剂对细胞内钙离子编织峰高度的影响也存在相关性。然而，每个细胞对激动剂的敏感性之间没有相关性，这是在加入激动剂后钙离子升高之前的潜伏期测量的。对细胞内几个小区域钙瞬变的动力学分析表明：(1)细胞内Ca~(2+)和Gt~(2+)波的空间分布模式不依赖于细胞局部1，4，5-三磷酸肌醇(Ins(1，4，5)P_3)的增加及其传播，而取决于Ins(1，4，5)P_3敏感的Ca^&lt；2+&gt；池的分布；(2)每个细胞对激动剂的敏感性与形成的Ins(1，4，5)P_3浓度密切相关。

项目成果

川西,徹: "細胞内カルシウムの動的観測" O plus E. 180. 83-90 (1994)

Kawanishi, Toru：“细胞内钙的动态观察”O plus E. 180. 83-90 (1994)

Takashi KATO,Kazuhiro TOYODA,Hisayuki OHATA,Kazutaka MOMOSE,Michihito TAKAHASHI and Toru KAWANISHI: "Kinetic Profiles of calcium waves in primary cultured hepatocytes revealed by rapid scanning confocal microscopy." Jpn.J.Pharmacol.67. 19P (1995)

Takashi KATO、Kazuhiro TOYODA、Hisayuki OHATA、Kazutaka MOMOSE、Michihito TAKAHASHI 和 Toru KAWANISHI：“通过快速扫描共聚焦显微镜揭示原代培养肝细胞中钙波的动力学特征。”

川西 徹: "細胞内カルシウムの動的観測" O plus E. 180. 83-91 (1994)

川西彻：《细胞内钙的动态观察》O plus E.180.83-91(1994)

Toru KAWANISHI,Takashi KATO,Hiroki ASOU,Chikako UNEYAMA,Kazuhiro TOYODA,Kazutaka MOMOSE,Michihito TAKAHASHI,and Yuzo HAYASHI: "Hepatocyte growth factor-induced calcium waves in hepatocytes as revealed with rapid scanning confocal microscopy." Cell Calcium

Toru KAWANISHI、Takashi KATO、Hiroki ASOU、Chikako UNEYAMA、Kazuhiro TOYODA、Kazutaka MOMOSE、Michihito TAKAHASHI 和 Yuzo HAYASHI：“通过快速扫描共聚焦显微镜揭示肝细胞生长因子诱导的肝细胞钙波。”

Toru KAWANISHI: "The relation between apontaneous calcium oscillations and cell proliferation in cultured smooth muscle cells" Jpm. J. Pharmacol. 65. 59-62 (1994)

Toru KAWANISHI：“培养的平滑肌细胞中自发钙振荡与细胞增殖之间的关系”Jpm。