Real-time monitoring of somatostatin receptor-cAMP signaling in live pituitary.
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.
Jacobs
Stefan
S
Institute of Pharmacology and Toxicology, University of Würzburg, 97078 Würzburg, Germany.
Calebiro
Davide
D
Nikolaev
Viacheslav O
VO
Lohse
Martin J
MJ
Schulz
Stefan
S
D000818:Animals; D002478:Cells, Cultured; D000242:Cyclic AMP / Q000378:metabolism*; D005260:Female; D031541:Fluorescence Resonance Energy Transfer / Q000379:methods; D020662:Guanine Nucleotide Exchange Factors / Q000235:genetics / Q000378:metabolism; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism; D008297:Male; D051379:Mice; D018345:Mice, Knockout; D008822:Mice, Transgenic; D056004:Molecular Dynamics Simulation; D010902:Pituitary Gland / Q000166:cytology / Q000378:metabolism*; D043562:Receptors, G-Protein-Coupled / Q000235:genetics / Q000378:metabolism; D017481:Receptors, Somatostatin / Q000235:genetics / Q000378:metabolism*; D015398:Signal Transduction; D013997:Time Factors
C482601:Epac protein, mouse; D020662:Guanine Nucleotide Exchange Factors; D008164:Luminescent Proteins; D043562:Receptors, G-Protein-Coupled; D017481:Receptors, Somatostatin; C492049:Sstr2 protein, mouse; D000242:Cyclic AMP