A Highly Sensitive A-Kinase Activity Reporter for Imaging Neuromodulatory Events in Awake Mice

Lei Ma, Bart C. Jongbloets, Wei-Hong Xiong, Joshua B. Melander, Maozhen Qin, Tess J. Lameyer, Madeleine F. Harrison, Boris V. Zemelman, Tianyi Mao, Haining Zhong

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

Neuromodulation imposes powerful control over brain function, and cAMP-dependent protein kinase (PKA) is a central downstream mediator of multiple neuromodulators. Although genetically encoded PKA sensors have been developed, single-cell imaging of PKA activity in living mice has not been established. Here, we used two-photon fluorescence lifetime imaging microscopy (2pFLIM) to visualize genetically encoded PKA sensors in response to the neuromodulators norepinephrine and dopamine. We screened available PKA sensors for 2pFLIM and further developed a variant (named tAKARα) with increased sensitivity and a broadened dynamic range. This sensor allowed detection of PKA activation by norepinephrine at physiologically relevant concentrations and kinetics, and by optogenetically released dopamine. In vivo longitudinal 2pFLIM imaging of tAKARα tracked bidirectional PKA activities in individual neurons in awake mice and revealed neuromodulatory PKA events that were associated with wakefulness, pharmacological manipulation, and locomotion. This new sensor combined with 2pFLIM will enable interrogation of neuromodulation-induced PKA signaling in awake animals. Video Abstract: It remains challenging to visualize neuromodulation, a major mode of neuronal communication, in vivo. Ma et al. demonstrate that two-photon fluorescence lifetime imaging of an improved genetically encoded sensor enables monitoring of neuromodulatory PKA signaling with single-cell resolution in behaving mice.

Original languageEnglish (US)
Pages (from-to)665-679.e5
JournalNeuron
Volume99
Issue number4
DOIs
StatePublished - Aug 22 2018

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Protein Kinases
Phosphotransferases
Optical Imaging
Photons
Neurotransmitter Agents
Dopamine
Norepinephrine
Wakefulness
Locomotion
Cyclic AMP-Dependent Protein Kinases
Microscopy
Communication
Pharmacology
Neurons
Brain

Keywords

  • A Kinase Activity Reporter (AKAR)
  • cAMP dependent kinase (PKA)
  • dopamine
  • enforced locomotion
  • Forster resonance energy transfer (FRET)
  • in vivo imaging
  • neuromodulation
  • norepinephrine
  • two-photon fluorescence lifetime imaging (2pFLIM)
  • wakefulness

ASJC Scopus subject areas

  • Neuroscience(all)

Cite this

A Highly Sensitive A-Kinase Activity Reporter for Imaging Neuromodulatory Events in Awake Mice. / Ma, Lei; Jongbloets, Bart C.; Xiong, Wei-Hong; Melander, Joshua B.; Qin, Maozhen; Lameyer, Tess J.; Harrison, Madeleine F.; Zemelman, Boris V.; Mao, Tianyi; Zhong, Haining.

In: Neuron, Vol. 99, No. 4, 22.08.2018, p. 665-679.e5.

Research output: Contribution to journalArticle

Ma, L, Jongbloets, BC, Xiong, W-H, Melander, JB, Qin, M, Lameyer, TJ, Harrison, MF, Zemelman, BV, Mao, T & Zhong, H 2018, 'A Highly Sensitive A-Kinase Activity Reporter for Imaging Neuromodulatory Events in Awake Mice', Neuron, vol. 99, no. 4, pp. 665-679.e5. https://doi.org/10.1016/j.neuron.2018.07.020
Ma, Lei ; Jongbloets, Bart C. ; Xiong, Wei-Hong ; Melander, Joshua B. ; Qin, Maozhen ; Lameyer, Tess J. ; Harrison, Madeleine F. ; Zemelman, Boris V. ; Mao, Tianyi ; Zhong, Haining. / A Highly Sensitive A-Kinase Activity Reporter for Imaging Neuromodulatory Events in Awake Mice. In: Neuron. 2018 ; Vol. 99, No. 4. pp. 665-679.e5.
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