A Cre-dependent reporter mouse for quantitative real-time imaging of protein kinase A activity dynamics
Abstract 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, FLI...
| Main Authors: | , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Nature Portfolio
2024-02-01
|
| Series: | Scientific Reports |
| Online Access: | https://doi.org/10.1038/s41598-024-53313-8 |
| _version_ | 1797275139086221312 |
|---|---|
| author | Elizabeth I. Tilden Aditi Maduskar Anna Oldenborg Bernardo L. Sabatini Yao Chen |
| author_facet | Elizabeth I. Tilden Aditi Maduskar Anna Oldenborg Bernardo L. Sabatini Yao Chen |
| author_sort | Elizabeth I. Tilden |
| collection | DOAJ |
| description | Abstract 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 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 Gαs and Gαq-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. |
| first_indexed | 2024-03-07T15:10:03Z |
| format | Article |
| id | doaj.art-58102d2e47ca4987be3fb89491c7361d |
| institution | Directory Open Access Journal |
| issn | 2045-2322 |
| language | English |
| last_indexed | 2024-03-07T15:10:03Z |
| publishDate | 2024-02-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Scientific Reports |
| spelling | doaj.art-58102d2e47ca4987be3fb89491c7361d2024-03-05T18:43:33ZengNature PortfolioScientific Reports2045-23222024-02-0114111110.1038/s41598-024-53313-8A Cre-dependent reporter mouse for quantitative real-time imaging of protein kinase A activity dynamicsElizabeth I. Tilden0Aditi Maduskar1Anna Oldenborg2Bernardo L. Sabatini3Yao Chen4Department of Neuroscience, Washington University in St. LouisDepartment of Neuroscience, Washington University in St. LouisDepartment of Neuroscience, Washington University in St. LouisDepartment of Neurobiology, Howard Hughes Medical Institute, Harvard Medical SchoolDepartment of Neuroscience, Washington University in St. LouisAbstract 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 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 Gαs and Gαq-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.https://doi.org/10.1038/s41598-024-53313-8 |
| spellingShingle | Elizabeth I. Tilden Aditi Maduskar Anna Oldenborg Bernardo L. Sabatini Yao Chen A Cre-dependent reporter mouse for quantitative real-time imaging of protein kinase A activity dynamics Scientific Reports |
| title | A Cre-dependent reporter mouse for quantitative real-time imaging of protein kinase A activity dynamics |
| title_full | A Cre-dependent reporter mouse for quantitative real-time imaging of protein kinase A activity dynamics |
| title_fullStr | A Cre-dependent reporter mouse for quantitative real-time imaging of protein kinase A activity dynamics |
| title_full_unstemmed | A Cre-dependent reporter mouse for quantitative real-time imaging of protein kinase A activity dynamics |
| title_short | A Cre-dependent reporter mouse for quantitative real-time imaging of protein kinase A activity dynamics |
| title_sort | cre dependent reporter mouse for quantitative real time imaging of protein kinase a activity dynamics |
| url | https://doi.org/10.1038/s41598-024-53313-8 |
| work_keys_str_mv | AT elizabethitilden acredependentreportermouseforquantitativerealtimeimagingofproteinkinaseaactivitydynamics AT aditimaduskar acredependentreportermouseforquantitativerealtimeimagingofproteinkinaseaactivitydynamics AT annaoldenborg acredependentreportermouseforquantitativerealtimeimagingofproteinkinaseaactivitydynamics AT bernardolsabatini acredependentreportermouseforquantitativerealtimeimagingofproteinkinaseaactivitydynamics AT yaochen acredependentreportermouseforquantitativerealtimeimagingofproteinkinaseaactivitydynamics AT elizabethitilden credependentreportermouseforquantitativerealtimeimagingofproteinkinaseaactivitydynamics AT aditimaduskar credependentreportermouseforquantitativerealtimeimagingofproteinkinaseaactivitydynamics AT annaoldenborg credependentreportermouseforquantitativerealtimeimagingofproteinkinaseaactivitydynamics AT bernardolsabatini credependentreportermouseforquantitativerealtimeimagingofproteinkinaseaactivitydynamics AT yaochen credependentreportermouseforquantitativerealtimeimagingofproteinkinaseaactivitydynamics |