Knock-in Luciferase Reporter Mice for In Vivo Monitoring of CREB Activity

• Published in: PloS one Vol. 11; no. 6; p. e0158274
• Main Authors: Akhmedov, Dmitry, Rajendran, Kavitha, Mendoza-Rodriguez, Maria G, Berdeaux, Rebecca
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 23.06.2016; Public Library of Science (PLoS)
• Subjects: Albinism; Animals; Biology; Biology and Life Sciences; Bioluminescence; Biosynthesis; Cloning; CREB (DNA binding protein); Cyclic AMP response element-binding protein; Dynamic tests; Expression vectors; Fasting; G proteins; Genes; Hepatocytes; Homeostasis; In vivo methods and tests; Influence; Kinases; Laboratory animals; Liver; Luciferase; Medical schools; Medicine and Health Sciences; Metabolism; Muscles; Musculoskeletal system; Myocytes; Neurosciences; Pharmacology; Physical Sciences; Physiology; Proteins; Research and Analysis Methods; Rodents; Science; Skeletal muscle; Transcription; Transgenes; Transgenic; United States > US; Texas
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0158274

The cAMP response element binding protein (CREB) is induced during fasting in the liver, where it stimulates transcription of rate-limiting gluconeogenic genes to maintain metabolic homeostasis. Adenoviral and transgenic CREB reporters have been used to monitor hepatic CREB activity non-invasively using bioluminescence reporter imaging. However, adenoviral vectors and randomly inserted transgenes have several limitations. To overcome disadvantages of the currently used strategies, we created a ROSA26 knock-in CREB reporter mouse line (ROSA26-CRE-luc). cAMP-inducing ligands stimulate the reporter in primary hepatocytes and myocytes from ROSA26-CRE-luc animals. In vivo, these animals exhibit little hepatic CREB activity in the ad libitum fed state but robust induction after fasting. Strikingly, CREB was markedly stimulated in liver, but not in skeletal muscle, after overnight voluntary wheel-running exercise, uncovering differential regulation of CREB in these tissues under catabolic states. The ROSA26-CRE-luc mouse line is a useful resource to study dynamics of CREB activity longitudinally in vivo and can be used as a source of primary cells for analysis of CREB regulatory pathways ex vivo.