Visualization of Cellular Functions

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Our primary goal is to better understand how the molecules for life behave in space and time. Signal transduction cascades involve multiple molecular components and are orchestrated by specific interactions and regulations. Such dynamics is revealed by optical means such as fluorescence readout. The green fluorescent protein (GFP) of thejelly fish Aequorea victoria is a spontaneously fluorescent, and has revolutionized our molecular and cell biology. Our projects mainly involve advancement of the GFP technologies, which include development ofgenetically-encodable indicators for specific cellular functions. One approach is to use twoGFPs of different colors to permit fluorescence resonance energy transfer (FRET), which is highly sensitive to the relative orientation and distance between the twofluorophores and alters the ratio of their emission intensities, an ideal readout for fast imaging and confocal microscopy. Another approach is to use circularly permuted GFP (cpGFP), in which the amino- and carboxy- portions have been interchanged and reconnected by a short spacer between the original termini. By exploiting these technologies, we have created not only calcium-sensitive proteins to obtain a better understanding of how intracellular calcium signals are generated and interpreted, but also new fluorescent indicators for visualization of signal transduction cascades that are currently assayed by grinding millions of cells. Furthermore, we develop new and unique optical technologies, by which fluorescent signals can be efficiently detected. For instance, our novel multi-color fluorescence imaging system allows the simultaneous observation of calcium-related events in conjunction with calcium dynamics in cells loaded with two or more fluorescent dyes.
日本組織細胞化学会の論文

Visualization of Cellular Functions

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