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      Imaging ERK and PKA Activation in Single Dendritic Spines during Structural Plasticity

      research-article
      1 , 1
      Neuron
      FRET, FLIM, synaptic plasticity, LTP, hippocampus, kinase, signaling

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          SUMMARY

          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.

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          Author and article information

          Journal
          8809320
          1600
          Neuron
          Neuron
          Neuron
          0896-6273
          1097-4199
          6 July 2018
          09 March 2017
          22 March 2017
          12 July 2018
          : 93
          : 6
          : 1315-1324.e3
          Affiliations
          [1 ]Max Planck Florida Institute for Neuroscience, 1 Max Planck Way, Jupiter, FL 33458, USA
          Author notes
          [* ]Correspondence: ryohei.yasuda@ 123456mpfi.org
          [2]

          Lead Contact

          Article
          PMC6042854 PMC6042854 6042854 nihpa979673
          10.1016/j.neuron.2017.02.032
          6042854
          28285819
          628ffe79-4359-49a6-a122-0ca6d234e36c
          History
          Categories
          Article

          signaling,FRET,FLIM,synaptic plasticity,LTP,hippocampus,kinase
          signaling, FRET, FLIM, synaptic plasticity, LTP, hippocampus, kinase

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