Gs-DREADD Knock-In Mice for Tissue-Specific, Temporal Stimulation of Cyclic AMP Signaling.
Hundreds of hormones and ligands stimulate cyclic AMP (cAMP) signaling in different tissues through the activation of G-protein-coupled receptors (GPCRs). Although the functions and individual effectors of cAMP signaling are well characterized in many tissues, pleiotropic effects of GPCR agonists limit investigations of physiological functions of cAMP signaling in individual cell types at different developmental stagesin vivoTo facilitate studies of cAMP signaling in specific cell populationsin vivo, we harnessed the power of DREADD (designer receptors exclusively activated by designer drugs) technology by creatingROSA26-based knock-in mice for the conditional expression of a Gs-coupled DREADD (rM3Ds-green fluorescent protein [GFP], or "GsD"). After Cre recombinase expression, GsD is activated temporally by the administration of the ligand clozapineN-oxide (CNO). In the same allele, we engineered a CREB-luciferase reporter transgene for noninvasive bioluminescence monitoring of CREB activity. After viral delivery of Cre recombinase to hepatocytesin vivo, GsD is expressed and allows CNO-dependent cAMP signaling and glycogen breakdown. The long-term expression of GsD in the liver results in constitutive CREB activity and hyperglycemia.ROSA26-Gs-DREADD mice can be used to study the physiological effects of cAMP signaling, acute or chronic, in liver or any tissue or cell type for which transgenic or viral Cre drivers are available.
Department of Integrative Biology and Pharmacology, McGovern Medical School at The University of Texas Health Science Center at Houston, Houston, Texas, USA.
Dmitry
D
Akhmedov
Department of Integrative Biology and Pharmacology, McGovern Medical School at The University of Texas Health Science Center at Houston, Houston, Texas, USA.
Maria G
MG
Mendoza-Rodriguez
Department of Integrative Biology and Pharmacology, McGovern Medical School at The University of Texas Health Science Center at Houston, Houston, Texas, USA.
Kavitha
K
Rajendran
Molecular Signaling Section, Laboratory of Bioorganic Chemistry, National Institute of Diabetes and Digestive and Kidney Diseases, Bethesda, Maryland, USA.
Mario
M
Rossi
Molecular Signaling Section, Laboratory of Bioorganic Chemistry, National Institute of Diabetes and Digestive and Kidney Diseases, Bethesda, Maryland, USA.
Jürgen
J
Wess
Department of Integrative Biology and Pharmacology, McGovern Medical School at The University of Texas Health Science Center at Houston, Houston, Texas, USA Rebecca.Berdeaux@uth.tmc.edu.|Institute of Molecular Medicine Center for Metabolic and Degenerative Diseases, McGovern Medical School at The University of Texas Health Science Center at Houston, Houston, Texas, USA.|Cell and Regulatory Biology Program of The University of Texas Graduate School of Biomedical Sciences, McGovern Medical School at The University of Texas Health Science Center at Houston, Houston, Texas, USA.
Rebecca
http://orcid.org/0000-0002-9929-1711
R
Berdeaux
D000818:Animals; D001786:Blood Glucose / Q000378:metabolism; D002478:Cells, Cultured; D003024:Clozapine / Q000031:analogs & derivatives / Q000494:pharmacology; D000242:Cyclic AMP / Q000378:metabolism*; D017362:Cyclic AMP Response Element-Binding Protein / Q000235:genetics; D005947:Glucose / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000235:genetics; D022781:Hepatocytes / Q000378:metabolism*; D006943:Hyperglycemia / Q000378:metabolism; D019426:Integrases / Q000235:genetics; D008099:Liver / Q000166:cytology / Q000378:metabolism*; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D022661:RNA, Untranslated / Q000235:genetics*; D043562:Receptors, G-Protein-Coupled / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D015398:Signal Transduction
D001786:Blood Glucose; C492004:Creb1 protein, mouse; D017362:Cyclic AMP Response Element-Binding Protein; C110853:Gt(ROSA)26Sor non-coding RNA, mouse; D022661:RNA, Untranslated; D043562:Receptors, G-Protein-Coupled; D049452:Green Fluorescent Proteins; D000242:Cyclic AMP; C045073:Cre recombinase; D019426:Integrases; D005947:Glucose; D003024:Clozapine; C079149:clozapine N-oxide
CREB; DREADD; GPCR; bioluminescence imaging; cyclic AMP; glucose metabolism; liver