1] Emmy Noether Group of the DFG, European Heart Research Institute Göttingen, University Medical Center Göttingen, D-37075 Göttingen, Germany [2] Department of Cardiology and Pulmonology, Heart Research Center Göttingen, University Medical Center Göttingen, Georg August University, D-37075 Göttingen, Germany [3] Institute of Experimental Cardiovascular Research, University Medical Center Hamburg-Eppendorf, D-20246 Hamburg, Germany.
Department of Cardiology and Pulmonology, Heart Research Center Göttingen, University Medical Center Göttingen, Georg August University, D-37075 Göttingen, Germany.
Nat Commun. 2015 Apr 28;6:6965. doi: 10.1038/ncomms7965.
3',5'-cyclic adenosine monophosphate (cAMP) is an ubiquitous second messenger that regulates physiological functions by acting in distinct subcellular microdomains. Although several targeted cAMP biosensors are developed and used in single cells, it is unclear whether such biosensors can be successfully applied in vivo, especially in the context of disease. Here, we describe a transgenic mouse model expressing a targeted cAMP sensor and analyse microdomain-specific second messenger dynamics in the vicinity of the sarcoplasmic/endoplasmic reticulum calcium ATPase (SERCA). We demonstrate the biocompatibility of this targeted sensor and its potential for real-time monitoring of compartmentalized cAMP signalling in adult cardiomyocytes isolated from a healthy mouse heart and from an in vivo cardiac disease model. In particular, we uncover the existence of a phosphodiesterase-dependent receptor-microdomain communication, which is affected in hypertrophy, resulting in reduced β-adrenergic receptor-cAMP signalling to SERCA.
3',5'-环磷酸腺苷 (cAMP) 是一种普遍存在的第二信使，通过在不同的亚细胞微域中发挥作用来调节生理功能。尽管已经开发并在单细胞中使用了几种靶向 cAMP 生物传感器，但尚不清楚这些生物传感器是否可以成功应用于体内，特别是在疾病的背景下。在这里，我们描述了一种表达靶向 cAMP 传感器的转基因小鼠模型，并分析了肌浆网/内质网钙 ATP 酶 (SERCA) 附近的微域特异性第二信使动力学。我们证明了这种靶向传感器的生物相容性及其在实时监测来自健康小鼠心脏和体内心脏疾病模型的分离成年心肌细胞中分隔的 cAMP 信号传导的潜力。特别是，我们揭示了存在一种依赖磷酸二酯酶的受体-微域通讯，该通讯在肥大中受到影响，导致 β-肾上腺素能受体-cAMP 信号传导到 SERCA 减少。
