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High‐resolution imaging of Ca 2+ , redox status, ROS and pH using GFP biosensors
Author(s) -
Choi WonGyu,
Swanson Sarah J.,
Gilroy Simon
Publication year - 2012
Publication title -
the plant journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.058
H-Index - 269
eISSN - 1365-313X
pISSN - 0960-7412
DOI - 10.1111/j.1365-313x.2012.04917.x
Subject(s) - reactive oxygen species , green fluorescent protein , biophysics , subcellular localization , biosensor , chemistry , redox , nanotechnology , biochemistry , biology , microbiology and biotechnology , materials science , cytoplasm , gene , organic chemistry
Summary Many plant response systems are linked to complex dynamics in signaling molecules such as Ca 2+ and reactive oxygen species (ROS) and to pH. Regulatory changes in these molecules can occur in the timeframe of seconds and are often limited to specific subcellular locales. Thus, to understand how Ca 2+ , ROS and pH form part of plants’ regulatory networks, it is essential to capture their rapid dynamics with resolutions that span the whole plant to subcellular dimensions. Defining the spatio‐temporal signaling ‘signatures’ of these regulators at high resolution has now been greatly facilitated by the generation of plants expressing a range of GFP‐based bioprobes. For Ca 2+ and pH, probes such as the yellow cameleon Ca 2+ sensors (principally YC2.1 and 3.6) or the pHluorin and H148D pH sensors provide a robust suite of tools to image changes in these ions. For ROS, the tools are much more limited, with the GFP‐based H 2 O 2 sensor Hyper representing a significant advance for the field. However, with this probe, its marked pH sensitivity provides a key challenge to interpretation without using appropriate controls to test for potentially coupled pH‐dependent changes. Most of these Ca 2+ ‐, ROS‐ and pH‐imaging biosensors are compatible with the standard configurations of confocal microscopes available to many researchers. These probes therefore represent a readily accessible toolkit to monitor cellular signaling. Their use does require appreciation of a minimal set of controls but these are largely related to ensuring that neither the probe itself nor the imaging conditions used perturb the biology of the plant under study.