In Vivo Imaging of the Coupling between Neuronal and CREB Activity in the Mouse Brain

Neuron. 2020 Mar 4;105(5):799-812.e5. doi: 10.1016/j.neuron.2019.11.028. Epub 2019 Dec 26.


Sensory experiences cause long-term modifications of neuronal circuits by modulating activity-dependent transcription programs that are vital for regulation of long-term synaptic plasticity and memory. However, it has not been possible to precisely determine the interaction between neuronal activity patterns and transcription factor activity. Here we present a technique using two-photon fluorescence lifetime imaging (2pFLIM) with new FRET biosensors to chronically image in vivo signaling of CREB, an activity-dependent transcription factor important for synaptic plasticity, at single-cell resolution. Simultaneous imaging of the red-shifted CREB sensor and GCaMP permitted exploration of how experience shapes the interplay between CREB and neuronal activity in the neocortex of awake mice. Dark rearing increased the sensitivity of CREB activity to Ca2+ elevations and prolonged the duration of CREB activation to more than 24 h in the visual cortex. This technique will allow researchers to unravel the transcriptional dynamics underlying experience-dependent plasticity in the brain.

Keywords: CREB; FLIM; FRET; in vivo imaging.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Calcium / metabolism*
  • Cyclic AMP Response Element-Binding Protein / metabolism*
  • Darkness
  • Fluorescence Resonance Energy Transfer
  • Mice
  • Neocortex / cytology
  • Neocortex / metabolism*
  • Neural Pathways
  • Neuronal Plasticity*
  • Neurons / cytology
  • Neurons / metabolism*
  • Photic Stimulation
  • Signal Transduction
  • Single-Cell Analysis
  • Somatosensory Cortex / cytology
  • Somatosensory Cortex / metabolism
  • Visual Cortex / cytology
  • Visual Cortex / metabolism


  • Creb1 protein, mouse
  • Cyclic AMP Response Element-Binding Protein
  • Calcium