Abstract
Fluorescence resonance energy transfer using genetically encoded biosensors has proven to be a powerful technique to monitor the spatiotemporal dynamics of cAMP signals stimulated by G(s)-coupled receptors in living cells. In contrast, real-time imaging of G(i)-mediated cAMP signals under native conditions remains challenging. Here, we describe the use of transgenic mice for cAMP imaging in living pituitary slices and primary pituitary cells. This technique can be widely used to assess the contribution of various pituitary receptors, including individual G(i) protein-coupled somatostatin receptors, to the regulation of cAMP levels under physiologically relevant settings.
MeSH terms
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Animals
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Cells, Cultured
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Cyclic AMP / metabolism*
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Female
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Fluorescence Resonance Energy Transfer / methods
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Guanine Nucleotide Exchange Factors / genetics
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Guanine Nucleotide Exchange Factors / metabolism
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Luminescent Proteins / genetics
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Luminescent Proteins / metabolism
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Male
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Mice
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Mice, Knockout
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Mice, Transgenic
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Molecular Dynamics Simulation
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Pituitary Gland / cytology
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Pituitary Gland / metabolism*
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Receptors, G-Protein-Coupled / genetics
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Receptors, G-Protein-Coupled / metabolism
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Receptors, Somatostatin / genetics
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Receptors, Somatostatin / metabolism*
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Signal Transduction
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Time Factors
Substances
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Epac protein, mouse
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Guanine Nucleotide Exchange Factors
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Luminescent Proteins
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Receptors, G-Protein-Coupled
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Receptors, Somatostatin
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Sstr2 protein, mouse
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Cyclic AMP