A genetically encoded fluorescent sensor of ERK activity.
Proc Natl Acad Sci U S A
; 105(49): 19264-9, 2008 Dec 09.
Article
em En
| MEDLINE
| ID: mdl-19033456
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.
Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Células Piramidais
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Sistema de Sinalização das MAP Quinases
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Transferência Ressonante de Energia de Fluorescência
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MAP Quinases Reguladas por Sinal Extracelular
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Corantes Fluorescentes
Limite:
Animals
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Humans
Idioma:
En
Ano de publicação:
2008
Tipo de documento:
Article