A genetically encoded fluorescent sensor of ERK activity
- 9 December 2008
- journal article
- Published by Proceedings of the National Academy of Sciences in Proceedings of the National Academy of Sciences of the United States of America
- Vol. 105 (49), 19264-19269
- https://doi.org/10.1073/pnas.0804598105
Abstract
The activity of the ERK has complex spatial and temporal dynamics that are important for the specificity of downstream effects. However, current biochemical techniques do not allow for the measurement of ERK signaling with fine spatiotemporal resolution. We developed a genetically encoded, FRET-based sensor of ERK activity (the extracellular signal-regulated kinase activity reporter, EKAR), optimized for signal-to-noise ratio and fluorescence lifetime imaging. EKAR selectively and reversibly reported ERK activation in HEK293 cells after epidermal growth factor stimulation. EKAR signals were correlated with ERK phosphorylation, required ERK activity, and did not report the activities of JNK or p38. EKAR reported ERK activation in the dendrites and nucleus of hippocampal pyramidal neurons in brain slices after theta-burst stimuli or trains of back-propagating action potentials. EKAR therefore permits the measurement of spatiotemporal ERK signaling dynamics in living cells, including in neuronal compartments in intact tissues.Keywords
This publication has 48 references indexed in Scilit:
- The Spread of Ras Activity Triggered by Activation of a Single Dendritic SpineScience, 2008
- Locally dynamic synaptic learning rules in pyramidal neuron dendritesNature, 2007
- Analyzing protein kinase dynamics in living cells with FRET reportersMethods, 2006
- Mechanisms of MAPK signalling specificityBiochemical Society Transactions, 2006
- Supersensitive Ras activation in dendrites and spines revealed by two-photon fluorescence lifetime imagingNature Neuroscience, 2006
- Insulin disrupts β-adrenergic signalling to protein kinase A in adipocytesNature, 2005
- Late-phase long-term potentiation: getting to the nucleusNature Reviews Neuroscience, 2005
- Applying spectral fingerprinting to the analysis of FRET imagesMicroscopy Research and Technique, 2004
- An improved cyan fluorescent protein variant useful for FRETNature Biotechnology, 2004
- A variant of yellow fluorescent protein with fast and efficient maturation for cell-biological applicationsNature Biotechnology, 2002