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首页> 外文期刊>Proceedings of the National Academy of Sciences of the United States of America >Imaging cellular signals in the heart in vivo: Cardiac expression of the high-signal Ca~(2+) indicator GCaMP2
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Imaging cellular signals in the heart in vivo: Cardiac expression of the high-signal Ca~(2+) indicator GCaMP2

机译：在体内对心脏中的细胞信号进行成像：高信号Ca〜（2+）指示剂GCaMP2的心脏表达

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摘要

Genetically encoded sensor proteins provide unique opportunities to advance the understanding of complex cellular interactions in physiologically relevant contexts; however, previously described sensors have proved to be of limited use to report cell signaling in vivo in mammals. Here, we describe an improved Ca~(2+) sensor, GCaMP2, its inducible expression in the mouse heart, and its use to examine signaling in heart cells in vivo. The high brightness and stability of GCaMP2 enable the measurement of myocyte Ca~(2+) transients in all regions of the beating mouse heart and prolonged pacing and mapping studies in isolated, perfused hearts. Trans-gene expression is efficiently temporally regulated in cardiomyo-cyte GCaMP2 mice, allowing recording of in vivo signals 4 weeks after transgene induction. High-resolution imaging of Ca~(2+) waves in GCaMP2-expressing embryos revealed key aspects of electrical conduction in the preseptated heart. At embryonic day (e.d.) 10.5, atrial and ventricular conduction occur rapidly, consistent with the early formation of specialized conduction pathways. However, conduction is markedly slowed through the atrioventricular canal in the e.d. 10.5 heart, forming the basis for an effective atrioventricular delay before development of the AV node, as rapid ventricular activation occurs after activation of the distal AV canal tissue. Consistent with the elimination of the inner AV canal muscle layer at e.d. 13.5, atrioventricular conduction through the canal was abolished at this stage. These studies demonstrate that GCaMP2 will have broad utility in the dissection of numerous complex cellular interactions in mammals, in vivo.
机译：遗传编码的传感器蛋白提供了独特的机会，可以促进在生理相关情况下对复杂细胞相互作用的理解。然而，先前描述的传感器已被证明在报告哺乳动物体内细胞信号传导方面用途有限。在这里，我们描述了一种改进的Ca〜（2+）传感器GCaMP2，其在小鼠心脏中的可诱导表达，以及其在体内检查心脏细胞中的信号传导的用途。 GCaMP2的高亮度和稳定性使其能够在跳动的小鼠心脏的所有区域中测量心肌细胞Ca〜（2+）的瞬变，并能在孤立的灌注心脏中进行长时间的起搏和作图研究。转基因的表达在心肌细胞GCaMP2小鼠中得到有效的时间调控，允许在转基因诱导后4周记录体内信号。对表达GCaMP2的胚胎中Ca〜（2+）波的高分辨率成像揭示了心脏前部传导的关键方面。在胚胎日（例如）10.5，心房和心室传导迅速发生，这与早期的专门传导通路形成一致。但是，通过房室管的传导明显减慢。 10.5心脏，为房室结发展之前有效的房室延迟奠定了基础，因为快速的心室激活发生在远端AV管组织激活之后。与在e.d处消除内AV管肌肉层一致13.5，在此阶段，通过导管的房室传导被取消。这些研究表明，GCaMP2在体内哺乳动物体内众多复杂细胞相互作用的解剖中将具有广泛的用途。

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