Imaging Ca2+ Nanosparks in Heart With a New Targeted Biosensor

doi:10.1161/CIRCRESAHA.114.302938

CORC > 北京大学 > 生命科学学院

	Imaging Ca2+ Nanosparks in Heart With a New Targeted Biosensor
	Shang, Wei ; Lu, Fujian ; Sun, Tao ; Xu, Jiejia ; Li, Lin-Lin ; Wang, Yanru ; Wang, Gang ; Chen, Liangyi ; Wang, Xianhua ; Cannell, Mark B. ; Wang, Shi-Qiang ; Cheng, Heping
刊名	circulation research
	2014
关键词	biosensing techniques calcium signaling excitation-contraction coupling junctin ryanodine receptor calcium release channel triadin RAT CARDIAC MYOCYTES RYANODINE RECEPTOR SARCOPLASMIC-RETICULUM CALCIUM TRANSIENTS INDUCTION DECAY RELEASE MUSCLE SPARKS CELLS TERMINATION
DOI	10.1161/CIRCRESAHA.114.302938
英文摘要	Rationale: In cardiac dyads, junctional Ca2+ directly controls the gating of the ryanodine receptors (RyRs), and is itself dominated by RyR-mediated Ca2+ release from the sarcoplasmic reticulum. Existing probes do not report such local Ca2+ signals because of probe diffusion, so a junction-targeted Ca2+ sensor should reveal new information on cardiac excitation-contraction coupling and its modification in disease states. Objective: To investigate Ca2+ signaling in the nanoscopic space of cardiac dyads by targeting a new sensitive Ca2+ biosensor (GCaMP6f) to the junctional space. Methods and Results: By fusing GCaMP6f to the N terminus of triadin 1 or junctin, GCaMP6f-triadin 1/junctin was targeted to dyadic junctions, where it colocalized with t-tubules and RyRs after adenovirus-mediated gene transfer. This membrane protein-tagged biosensor displayed approximate to 4x faster kinetics than native GCaMP6f. Confocal imaging revealed junctional Ca2+ transients (Ca2+ nanosparks) that were approximate to 50x smaller in volume than conventional Ca2+ sparks (measured with diffusible indicators). The presence of the biosensor did not disrupt normal Ca2+ signaling. Because no indicator diffusion occurred, the amplitude and timing of release measurements were improved, despite the small recording volume. We could also visualize coactivation of subclusters of RyRs within a single junctional region, as well as quarky Ca2+ release events. Conclusions: This new, targeted biosensor allows selective visualization and measurement of nanodomain Ca2+ dynamics in intact cells and can be used to give mechanistic insights into dyad RyR operation in health and in disease states such as when RyRs become orphaned.; http://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcApp=PARTNER_APP&SrcAuth=LinksAMR&KeyUT=WOS:000335586800006&DestLinkType=FullRecord&DestApp=ALL_WOS&UsrCustomerID=8e1609b174ce4e31116a60747a720701 ; Cardiac & Cardiovascular Systems; Hematology; Peripheral Vascular Disease; SCI(E); 12; ARTICLE; wsq@pku.edu.cn; chengp@pku.edu.cn; 3; 412-420; 114
语种	英语
内容类型	期刊论文
源URL	[http://ir.pku.edu.cn/handle/20.500.11897/390497]
专题	生命科学学院
推荐引用方式 GB/T 7714	Shang, Wei,Lu, Fujian,Sun, Tao,et al. Imaging Ca2+ Nanosparks in Heart With a New Targeted Biosensor[J]. circulation research,2014.
APA	Shang, Wei.,Lu, Fujian.,Sun, Tao.,Xu, Jiejia.,Li, Lin-Lin.,...&Cheng, Heping.(2014).Imaging Ca2+ Nanosparks in Heart With a New Targeted Biosensor.circulation research.
MLA	Shang, Wei,et al."Imaging Ca2+ Nanosparks in Heart With a New Targeted Biosensor".circulation research (2014).