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

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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.