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

Authors:
Lei Ma
Lei Ma
Xijing Hospital
China
Bart C Jongbloets
Bart C Jongbloets
University Medical Center Utrecht
Netherlands
Wei-Hong Xiong
Wei-Hong Xiong
Johns Hopkins University School of Medicine
United States
Joshua B Melander
Joshua B Melander
Oregon Health and Science University
Maozhen Qin
Maozhen Qin
Vollum Institute
Tess J Lameyer
Tess J Lameyer
Madison | United States
Boris V Zemelman
Boris V Zemelman
Howard Hughes Medical Institute
United States

Neuron 2018 Aug 9;99(4):665-679.e5. Epub 2018 Aug 9.

Vollum Institute, Oregon Health & Science University, Portland, OR 97239, USA. Electronic address:

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.

Abstract Video

Seeing the Intracellular Action of Neuromodulation In Vivo


Source: Cell Press

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http://dx.doi.org/10.1016/j.neuron.2018.07.020DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6152931PMC

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August 2018
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