title: Imaging ERK and PKA Activation in Single Dendritic Spines during Structural Plasticity.
abstract: Extracellular signal-regulated kinase (ERK) and protein kinase A (PKA) play important roles in LTP and spine structural plasticity. While fluorescence resonance energy transfer (FRET)-based sensors for these kinases had previously been developed, they did not provide sufficient sensitivity for imaging small neuronal compartments, such as single dendritic spines in brain slices. Here we improved the sensitivity of FRET-based kinase sensors for monitoring kinase activity under two-photon fluorescence lifetime imaging microscopy (2pFLIM). Using these improved sensors, we succeeded in imaging ERK and PKA activation in single dendritic spines during structural long-term potentiation (sLTP) in hippocampal CA1 pyramidal neurons, revealing that the activation of these kinases spreads widely with length constants of more than 10 μm. The strategy for improvement of sensors used here should be applicable for developing highly sensitive biosensors for various protein kinases. VIDEO ABSTRACT.
authors.affiliation: Max Planck Florida Institute for Neuroscience, 1 Max Planck Way, Jupiter, FL 33458, USA.
authors.forename: Shen
authors.initials: S
authors.lastname: Tang
authors.affiliation: Max Planck Florida Institute for Neuroscience, 1 Max Planck Way, Jupiter, FL 33458, USA. Electronic address: ryohei.yasuda@mpfi.org.
authors.forename: Ryohei
authors.initials: R
authors.lastname: Yasuda
mesh_terms: D000818:Animals; D056547:CA1 Region, Hippocampal / Q000378:metabolism; D002478:Cells, Cultured; D017868:Cyclic AMP-Dependent Protein Kinases / Q000378:metabolism*; D049229:Dendritic Spines / Q000201:enzymology* / Q000502:physiology*; D048049:Extracellular Signal-Regulated MAP Kinases / Q000378:metabolism*; D005260:Female; D031541:Fluorescence Resonance Energy Transfer / Q000379:methods*; D006801:Humans; D017774:Long-Term Potentiation / Q000502:physiology*; D008297:Male; D051379:Mice; D036641:Microscopy, Fluorescence, Multiphoton; D009473:Neuronal Plasticity / Q000502:physiology*; D017966:Pyramidal Cells / Q000378:metabolism
chemical_list: D017868:Cyclic AMP-Dependent Protein Kinases; D048049:Extracellular Signal-Regulated MAP Kinases
keywords: FLIM; FRET; LTP; hippocampus; kinase; signaling; synaptic plasticity