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

被引：55

作者：

Ma, Lei ^{[1
]}

Jongbloets, Bart C. ^{[1
]}

Xiong, Wei-Hong ^{[1
]}

Melander, Joshua B. ^{[1
,3
]}

Qin, Maozhen ^{[1
]}

Lameyer, Tess J. ^{[1
]}

Harrison, Madeleine F. ^{[2
]}

Zemelman, Boris V. ^{[2
]}

Mao, Tianyi ^{[1
]}

Zhong, Haining ^{[1
]}

机构：

[1] Oregon Hlth & Sci Univ, Vollum Inst, Portland, OR 97239 USA

[2] Univ Texas Austin, Ctr Learning & Memory, Austin, TX 78712 USA

[3] Stanford Univ, Stanford Neurosci Program, Stanford, CA 94305 USA

来源：

NEURON | 2018年 / 99卷 / 04期

关键词：

IN-VIVO; DEVELOPMENTAL SWITCH; SUBCELLULAR DYNAMICS; PKA; DOPAMINE; CAMP; NORADRENALINE; MODULATION; NOREPINEPHRINE; MECHANISMS;

D O I：

10.1016/j.neuron.2018.07.020

中图分类号：

Q189 [神经科学];

学科分类号：

071006 ;

摘要：

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 tAKAR alpha) 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 tAKARa 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.

引用

页码：665 / +

页数：20