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.
Copyright © 2019 Elsevier Inc. All rights reserved.
Conflict of interest statement
Declaration of Interests R.Y. is a founder of Florida Lifetime Imaging LLC.
The Role of CREB, SRF, and MEF2 in Activity-Dependent Neuronal Plasticity in the Visual Cortex.J Neurosci. 2017 Jul 12;37(28):6628-6637. doi: 10.1523/JNEUROSCI.0766-17.2017. Epub 2017 Jun 12. J Neurosci. 2017. PMID: 28607167 Free PMC article.
Activity-Dependent Dynamics of the Transcription Factor of cAMP-Response Element Binding Protein in Cortical Neurons Revealed by Single-Molecule Imaging.J Neurosci. 2017 Jan 4;37(1):1-10. doi: 10.1523/JNEUROSCI.0943-16.2016. J Neurosci. 2017. PMID: 28053025 Free PMC article.
Shaping synaptic plasticity: the role of activity-mediated epigenetic regulation on gene transcription.Int J Dev Neurosci. 2013 Oct;31(6):359-69. doi: 10.1016/j.ijdevneu.2013.04.003. Epub 2013 May 9. Int J Dev Neurosci. 2013. PMID: 23665156 Review.
Brief Novel Visual Experience Fundamentally Changes Synaptic Plasticity in the Mouse Visual Cortex.J Neurosci. 2017 Sep 27;37(39):9353-9360. doi: 10.1523/JNEUROSCI.0334-17.2017. Epub 2017 Aug 18. J Neurosci. 2017. PMID: 28821676 Free PMC article.
Visualizing Adult Cortical Plasticity Using Intrinsic Signal Optical Imaging.In: Frostig RD, editor. In Vivo Optical Imaging of Brain Function. 2nd edition. Boca Raton (FL): CRC Press/Taylor & Francis; 2009. Chapter 9. In Vivo Optical Imaging of Brain Function. 2nd edition. 2009. PMID: 26844321 Free Books & Documents. Review.