Gs-DREADD Knock-In Mice for Tissue-Specific, Temporal Stimulation of Cyclic AMP Signaling.
Gs-DREADD Knock-In Mice for Tissue-Specific, Temporal Stimulation of Cyclic AMP Signaling.
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DOI：
10.1128/mcb.00584-16
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发表时间：
2017-05-01
影响因子：
5.3
通讯作者：
Berdeaux R
中科院分区：
文献类型：
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作者：
Akhmedov D;Mendoza-Rodriguez MG;Rajendran K;Rossi M;Wess J;Berdeaux R
数以百计的激素和配体通过激活g蛋白偶联受体（gpcr）刺激不同组织中的环AMP （cAMP）信号。尽管cAMP信号的功能和个体效应在许多组织中都得到了很好的表征，但GPCR激动剂的多效性限制了对体内不同发育阶段个体细胞类型中cAMP信号生理功能的研究。为了促进体内特定细胞群中cAMP信号的研究，我们利用了DREADD（由设计药物完全激活的设计受体）技术的力量，通过创建基于rosa26的敲入小鼠来条件表达gs偶联的DREADD （rm3d -绿色荧光蛋白[GFP]，或“GsD”）。Cre重组酶表达后，GsD被配体氯氮平n -氧化物（CNO）暂时激活。在相同的等位基因中，我们设计了一个CREB荧光素酶报告基因，用于CREB活性的无创生物发光监测。在体内，病毒将Cre重组酶传递到肝细胞后，GsD被表达，并允许cno依赖性cAMP信号和糖原分解。GsD在肝脏中的长期表达导致构成性CREB活性和高血糖。ROSA26-Gs-DREADD小鼠可用于研究急性或慢性cAMP信号在肝脏或任何可获得转基因或病毒Cre驱动的组织或细胞类型中的生理作用。
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 stages in vivo. To facilitate studies of cAMP signaling in specific cell populations in vivo, we harnessed the power of DREADD (designer receptors exclusively activated by designer drugs) technology by creating ROSA26-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 clozapine N-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 hepatocytes in 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.
DOI：
10.1007/978-1-4939-2914-6_14
发表时间：
2015
期刊：
Methods in molecular biology (Clifton, N.J.)
影响因子：
--
作者：
Rossi, Mario;Cui, Zhenzhong;Nakajima, Ken-ichiro;Hu, Jianxin;Zhu, Lu;Wess, Jurgen
通讯作者：
Wess, Jurgen