Abstract
AbstractIntracellular signaling dynamics play a crucial role in cell function. Protein kinase A (PKA) is a key signaling molecule that has diverse functions, from regulating metabolism and brain activity to guiding development and cancer progression. We previously developed an optical reporter, FLIM-AKAR, that allows for quantitative imaging of PKA activity via fluorescence lifetime imaging microscopy and photometry. However, using viral infection or electroporation for the delivery of FLIM-AKAR is invasive, cannot easily target sparse or hard-to-transfect/infect cell types, and results in variable expression. Here, we developed a reporter mouse,FL-AK, which expresses FLIM-AKAR in aCre-dependent manner from theROSA26locus.FL-AKprovides robust and consistent expression of FLIM-AKAR over time. Functionally, the mouse line reports an increase in PKA activity in response to activation of both Gαsand Gαq-coupled receptors in brain slices.In vivo, FL-AKreports PKA phosphorylation in response to neuromodulator receptor activation. Thus,FL-AKprovides a quantitative, robust, and flexible method to reveal the dynamics of PKA activity in diverse cell types.
Publisher
Cold Spring Harbor Laboratory
Cited by
1 articles.
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