Highly dynamic and sensitive NEMOer calcium indicators for imaging ER calcium signals in excitable cells

Wenjia Gu, Jia-Hui Chen, Yiyin Zhang, Zhirong Wang, Jia Li, Sijia Wang, Hanhan Zhang, Amin Jiang, Ziyi Zhong, Jiaxuan Zhang, Ze Xu, Panpan Liu, Chao Xi, Tingting Hou, Donald L. Gill, Dong Li, Yu Mu (), Shi-Qiang Wang (), Ai-Hui Tang () and Youjun Wang ()
Additional contact information
Wenjia Gu: Beijing Normal University
Jia-Hui Chen: Anhui Medical University
Yiyin Zhang: Peking University
Zhirong Wang: Chinese Academy of Sciences
Jia Li: Beijing Normal University
Sijia Wang: Chinese Academy of Sciences
Hanhan Zhang: Beijing Normal University
Amin Jiang: Chinese Academy of Sciences
Ziyi Zhong: Beijing Normal University
Jiaxuan Zhang: Beijing Normal University
Ze Xu: Peking University
Panpan Liu: Beijing Normal University
Chao Xi: Beijing Normal University
Tingting Hou: Peking University
Donald L. Gill: Pennsylvania State University College of Medicine
Dong Li: Chinese Academy of Sciences
Yu Mu: Chinese Academy of Sciences
Shi-Qiang Wang: Peking University
Ai-Hui Tang: Hefei Comprehensive National Science Center
Youjun Wang: Beijing Normal University

Nature Communications, 2025, vol. 16, issue 1, 1-12

Abstract: Abstract The Endoplasmic/sarcoplasmic reticulum (ER/SR) is central to calcium (Ca2+) signaling, yet current genetically encoded Ca2+ indicators (GECIs) cannot detect elementary Ca2+ release events from ER/SR, particularly in muscle cells. Here, we report NEMOer, a set of organellar GECIs, to efficiently capture ER Ca2+ dynamics with increased sensitivity and responsiveness. NEMOer indicators exhibit dynamic ranges an order of magnitude larger than G-CEPIA1er, enabling 2.7-fold more sensitive detection of Ca2+ transients in both non-excitable and excitable cells. The ratiometric version further allows super-resolution monitoring of local ER Ca2+ homeostasis and dynamics. Notably, NEMOer-f enabled the inaugural detection of Ca2+ blinks, elementary Ca2+ releasing signals from the SR of cardiomyocytes, as well as in vivo spontaneous SR Ca2+ releases in zebrafish. In summary, the highly dynamic NEMOer sensors expand the repertoire of organellar Ca2+ sensors that allow real-time monitoring of intricate Ca2+ dynamics and homeostasis in live cells with high spatiotemporal resolution.

Date: 2025
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DOI: 10.1038/s41467-025-58705-6

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