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

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

doi:10.1016/j.neuron.2018.07.020

Cited by 70 publications

(68 citation statements)

References 86 publications

(118 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Direct measurements of the downstream effects of RPE-encoding DA transients on SPNs have been difficult to achieve due the challenges of monitoring intracellular signaling in behaving animals (although see Goto et al 46 , Yamaguchi et al 47 , and Ma et al 48 ). Employing the fluorescence lifetime photometry technique in combination of pharmacological and optogenetic manipulations, we successfully monitored the dynamics of net PKA activity in SPNs during behavior.…”

Section: Spn Pka Modulation and Function During Behaviormentioning

confidence: 99%

Cell-type specific asynchronous modulation of PKA by dopamine during reward based learning

Lee¹,

Lodder²,

Chen³

et al. 2019

Preprint

View full text Add to dashboard Cite

Canonical reinforcement learning models postulate that dopamine neurons encode reward prediction error (RPE) and provide a teaching signal to striatal spiny projection neurons (SPNs) in the form of dopamine (DA) release. DA is thought to guide learning via dynamic modulation of protein kinase A (PKA) in SPNs. However, this fundamental assumption remains untested in behaving animals. Here we utilized multi-channel fiber photometry and fluorescence lifetime photometry (FLiP) to monitor the activity of DA neurons, extracellular DA levels, and net PKA activity in SPNs in the nucleus accumbens during learning. We found dynamic encoding of RPE in the activity of DA neurons, which is both necessary and sufficient to explain striatal DA levels and SPN PKA activity. The modulation of PKA in SPNs that express type-1 (D1R-SPNs) and type-2 (D2R-SPNs) DA receptors was dichotomous such that in each cell class it is selectively sensitive to increases and decreases in DA, respectively, and occur at and support different phases of learning. Thus, PKA-dependent pathways in D1R-and D2R-SPNs are asynchronously engaged by RPE-encoding DA signals to promote different aspects of reinforcement learning: the former responsible for the initial association between action and outcome and the latter responsible for refining the learned association.

show abstract

Section: Spn Pka Modulation and Function During Behaviormentioning

confidence: 99%

Cell-type specific asynchronous modulation of PKA by dopamine during reward based learning

Lee¹,

Lodder²,

Chen³

et al. 2019

Preprint

View full text Add to dashboard Cite

show abstract

“…In this work, we used previously published FRET-based reporters for PKA and ERK1/2 referred to as AKAR and EKAR respectively 13 . Variants of these biosensors have previously been used in vivo with microendoscopy 3,13 , 2-photon microscopy 5 , and fluorescence lifetime photometry 4 . These biosensors are substrates for protein kinase activity and contain phosphorylation-dependent interaction domains such that the biosensor undergoes a conformational change upon phosphorylation, leading to an increase in resonance-energy transfer (Fig.…”

Section: Resultsmentioning

confidence: 99%

“…In vivo imaging of FRET biosensors has previously been accomplished using microendoscopy 3,14 , 2-photon microscopy 5 and fluorescence lifetime photometry 4 , but these techniques are costly, technically demanding and require specialized equipment that make them impractical for wide application in non-specialized labs. In contrast, fiber photometry is a simple method for performing functional fluorescence recording in vivo and is compatible with many common behavioral tests 7–9 .…”

Section: Resultsmentioning

confidence: 99%

“…Many intracellular signalling biosensors have been developed which utilize changes in Förster resonance energy transfer (FRET) between two fluorescent proteins 11 , in order to report levels or activity of second messengers, protein kinases, GTPases, post-translational modifications and protein-protein interactions 12–15 . These tools have been widely used to dissect signaling pathways in cultured cells and have recently begun to be applied in vivo , aided by recent technological developments such as intravital imaging 14,16 , microendoscopy 3 , 2-photon microscopy 2,5,6,17 and fluorescence lifetime measurement 4 . Here we report a ratiometric fiber-photometry approach for real-time recording of genetically-encoded FRET biosensors in freely moving rodents.…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Dopamine D1 receptor signalling in dyskinetic Parkinsonian rats revealed by fiber photometry using FRET-based biosensors

Jones-Tabah¹,

Mohammad²,

Hadj-Youssef³

et al. 2020

Preprint

View full text Add to dashboard Cite

Like many G protein-coupled receptors (GPCRs), the signalling pathways regulated by the dopamine D1 receptor (D1R) are dynamic, cell-type specific, and can change in response to disease or drug exposures. In striatal neurons, the D1R activates cAMP/protein kinase A (PKA) signalling. However, in Parkinson’s disease (PD), alterations in this pathway lead to activation of extracellular regulated kinases (ERK1/2), contributing to L-DOPA-induced dyskinesia (LID). In order to detect D1R activation in vivo and to study the progressive dysregulation of D1R signalling in PD and LID, we developed ratiometric fiber-photometry with Förster resonance energy transfer (FRET) biosensors and optically detected PKA and ERK1/2 signalling in freely moving rats. We show that in Parkinsonian animals, D1R signalling through PKA and ERK1/2 is sensitized, but that following chronic treatment with L-DOPA, these pathways become partially desensitized while concurrently D1R activation leads to greater induction of dyskinesia.

show abstract

“…Finally, further research is required to monitor cAMP production and PKA activation in behaving animals. Recent advances in the development of reporters for cAMP (Muntean et al, 2018), and PKA (Ma et al, 2018) will enable a spatial and temporal analysis of cAMP/PKA signaling in animals consuming alcohol.…”

Section: And the Collapsin Responsementioning

confidence: 99%

cAMP-Fyn signaling in the dorsomedial striatum direct pathway drives excessive alcohol use

Ehinger

Morisot

Phamluong

et al. 2020

Preprint

View full text Add to dashboard Cite

Fyn kinase in the dorsomedial striatum (DMS) of rodents plays a central role in mechanisms underlying excessive alcohol intake. The DMS is comprised mostly of medium spiny neurons (MSNs) that are divided into dopamine D1 receptor expressing neurons (dMSNs), and dopamine D2 receptor expressing neurons (iMSNs). Here, we examined the cell-type specificity of Fyn's actions on the development of alcohol use. First, we knocked down Fyn selectively in DMS dMSNs or iMSNs of mice and measured the level of alcohol consumption. We found that downregulation of Fyn in dMSNs, but not in iMSNs, reduces excessive alcohol but not saccharin intake and preference. D1Rs are coupled to Gαs/olf, which activate cAMP signaling. To examine whether Fyn's actions are mediated through cAMP signaling, DMS dMSNs were infected with GαsDREADD, and the activation of Fyn signaling was measured following CNO treatment. We found that remote stimulation of cAMP signaling in DMS dMSNs activates Fyn and promotes the phosphorylation of the Fyn substrate, GluN2B. Next, we tested whether remote activation of GαsDREADD in DMS dMSNs or iMSNs alters alcohol intake, and observed that CNO-dependent activation of GαsDREADD in DMS dMSNs but not iMSNs increases alcohol but not saccharin intake and preference. Finally, we examined the contribution of Fyn to GαsDREADD-dependent increase in alcohol intake, and found that systemic administration of the Fyn inhibitor, AZD0503 blocks GαsDREADD-dependent increase in alcohol consumption.Our results suggest that the cAMP-Fyn axis in the DMS dMSNs is a molecular transducer of mechanisms underlying the development of excessive alcohol consumption.

show abstract

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

Cited by 70 publications

References 86 publications

Cell-type specific asynchronous modulation of PKA by dopamine during reward based learning

Cell-type specific asynchronous modulation of PKA by dopamine during reward based learning

Dopamine D1 receptor signalling in dyskinetic Parkinsonian rats revealed by fiber photometry using FRET-based biosensors

cAMP-Fyn signaling in the dorsomedial striatum direct pathway drives excessive alcohol use

Contact Info

Product

Resources

About