Measurement of intracellular signaling events induced by cell-cell interactions in a live organ.

测量活体器官中细胞间相互作用诱导的细胞内信号传导事件。

• 批准号: 06557024
• 负责人: NAKAYAMA Toshinori
• 金额: $ 4.54万
• 依托单位: Science University of Tokyo
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (A)
• 财政年份: 1994
• 资助国家: 日本
• 起止时间: 1994 至 1996
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-06557024/
• 关键词: calcium; oscillation; コンピューター画像解析; カルシウムイオン; 刺激伝達系; リンパ球; カルシウムオシレーション; コンピュータ画像解析

We have tried to established a system, in which intracellular signaling event are quantitatively detected and visualized on the computer displays. The materials used are lymphocytes, thymocytes, and thymic organs. Those cells were atimulated with anti-T cell antigen receptor antibodies or PMA & ionomycin. The intra-cellular free calcium ion concentration ([Ca2+]i) was measured in the stimulated cells for 15 min.to 1 hour. We could detect a rapid increase and long-lasting oscillations of [Ca2+]i quantitatively. These data obtained by using this grant are published in several journals. The measurement was also succeeded in thymocytes presented in the organ cultured thymus. This assasy system is one of the novel experimental detection system, and provide a possibility for analyzing intra-cellular signaling events in live organs.

我们试图建立一种对细胞内信号事件进行定量检测并在计算机显示器上可视化的系统。使用的材料是淋巴细胞、胸腺细胞和胸腺器官。用抗T细胞抗原受体抗体或PMA和离子霉素刺激这些细胞。刺激15min~1h后，测定细胞内游离钙离子浓度([Ca~(2+)]i)。我们可以定量检测到[Ca~(2+)]i的快速升高和持续振荡。通过使用这笔赠款获得的这些数据发表在几个期刊上。在器官培养的胸腺中出现的胸腺细胞也成功地进行了测量。该系统是一种新颖的实验检测系统，为分析活体器官的细胞内信号事件提供了可能。

项目成果

Kearse, K.P., Roberts, J.L., Munitz, T.I., Wiest, D.L., Nakayama, T., and Singer A.: "Developmental regulation of alphabeta T cell antigen receptor expression results from differential stability of nascent TCRalpha proteins within the endoplasmic reticulu

Kearse, K.P.、Roberts, J.L.、Munitz, T.I.、Wiest, D.L.、Nakayama, T. 和 Singer A.：“alphata T 细胞抗原受体表达的发育调节是由内质网内新生 TCRalpha 蛋白的稳定性差异造成的。

Wange,C.R.: "T-Cell Receptor-Mediated Signaling Events in CD4^+CD8^+ Thymocytes Undergoing Thymic Selection - Requirement of Calcineurin Activation for Thymic Positive Selection but not Negative Selection" J.Exp.Med.181. 927-941 (1995)

Wange,C.R.：“进行胸腺选择的 CD4^ CD8^ 胸腺细胞中 T 细胞受体介导的信号转导事件 - 胸腺正选择而非负选择需要钙调神经磷酸酶激活”J.Exp.Med.181。

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