Knock-in Luciferase Reporter Mice for In Vivo Monitoring of CREB Activity.
Knock-in Luciferase Reporter Mice for In Vivo Monitoring of CREB Activity.
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DOI：
10.1371/journal.pone.0158274
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发表时间：
2016
期刊：
影响因子：
3.7
通讯作者：
Berdeaux R
中科院分区：
文献类型：
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作者：
Akhmedov D;Rajendran K;Mendoza-Rodriguez MG;Berdeaux R
CAMP反应元件结合蛋白(CREB)是在肝脏禁食期间诱导的，它刺激限速糖异生基因的转录，以维持代谢动态平衡。腺病毒和转基因CREB报告基因已被用于无创监测肝脏CREB活性的生物发光报告成像。然而，腺病毒载体和随机插入的转基因有几个局限性。为了克服目前使用的策略的缺点，我们创建了一个rosa26敲入CREB报告鼠系(rosa26-cre-Luc)。CAMP诱导配体在rosa26-Cre-Luc动物的原代肝细胞和心肌细胞中刺激报告基因。在活体内，这些动物在随意喂养状态下肝脏CREB活性很低，但禁食后表现出强劲的诱导活性。值得注意的是，经过整夜的自愿轮式运动后，CREB在肝脏中被显著刺激，但在骨骼肌中没有被刺激，揭示了CREB在分解代谢状态下在这些组织中的差异调节。ROSA26-Cre-Luc小鼠系是纵向研究体内CREB活性动态的有用资源，可作为体外CREB调控途径分析的原代细胞来源。
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.