The Theodore Bücher lecture. Investigating signal transduction with genetically encoded fluorescent probes.
Ca2+ and cAMP are ubiquitous second messengers in eukaryotes and control numerous physiological responses ranging from fertilization to cell death induction. To distinguish between these different responses, their subtle regulation in time, space and amplitude is needed. Therefore, the characterization of the signalling process requires measurement of second messengers with tools of precise localization, high dynamic range and as little disturbance of cell physiology as possible. Recently, fluorescent proteins of marine jellyfish have given rise to a set of genetically encoded biosensors which fulfil these criteria and which have already led to important new insights into the subcellular handling of Ca2+ and cAMP. The use of these probes in combination with new microscopical methods such as two-photon microscopy now enables researchers to study second messenger signalling in intact tissues. In this review, the genetically encoded measurement probes and their origin are briefly introduced and some recent insights into the spatio-temporal complexity of both Ca2+ and cAMP signalling obtained with these tools are discussed.
Pozzan
Tullio
T
Department of Biomedical Sciences, CNR Institute of Neurosciences, University of Padua, and Venetian Institute for Molecular Medicine, Padua, Italy. tullio.pozzan@unipd.it
Mongillo
Marco
M
Rudolf
Rüdiger
R
D000331:Aequorin / Q000378:metabolism; D000818:Animals; D015374:Biosensing Techniques; D002118:Calcium / Q000378:metabolism; D000242:Cyclic AMP / Q000378:metabolism; D004721:Endoplasmic Reticulum / Q000378:metabolism; D005456:Fluorescent Dyes / Q000494:pharmacology*; D049452:Green Fluorescent Proteins; D006801:Humans; D007700:Kinetics; D008164:Luminescent Proteins / Q000378:metabolism; D008928:Mitochondria / Q000378:metabolism; D008954:Models, Biological; D009154:Mutation; D011493:Protein Kinase C / Q000378:metabolism; D064546:Protein Kinase C beta; D037401:Scyphozoa; D015398:Signal Transduction; D014162:Transfection
D005456:Fluorescent Dyes; D008164:Luminescent Proteins; D049452:Green Fluorescent Proteins; D000331:Aequorin; D000242:Cyclic AMP; D011493:Protein Kinase C; D064546:Protein Kinase C beta; D002118:Calcium