First Author | Tilden EI | Year | 2023 |
Journal | bioRxiv | PubMed ID | 37961214 |
Mgi Jnum | J:343075 | Mgi Id | MGI:7563786 |
Doi | 10.1101/2023.10.31.565028 | Citation | Tilden EI, et al. (2023) A Cre-dependent reporter mouse for quantitative real-time imaging of Protein Kinase A activity dynamics. bioRxiv |
abstractText | Intracellular 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 a Cre-dependent manner from the ROSA26 locus. FL-AK provides 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 G(alphas) and G(alphaq)-coupled receptors in brain slices. In vivo, FL-AK reports PKA phosphorylation in response to neuromodulator receptor activation. Thus, FL-AK provides a quantitative, robust, and flexible method to reveal the dynamics of PKA activity in diverse cell types. |