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

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 efficie...

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Main Authors: 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, Youjun Wang
Format: Article
Language:English
Published: Nature Portfolio 2025-04-01
Series:Nature Communications
Online Access:https://doi.org/10.1038/s41467-025-58705-6
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author 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
Youjun Wang
author_facet 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
Youjun Wang
author_sort Wenjia Gu
collection DOAJ
description 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.
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issn 2041-1723
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publishDate 2025-04-01
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spelling doaj-art-dac6bfe4fd3b4d2a9dcfad7a1631a0e22025-08-20T02:11:42ZengNature PortfolioNature Communications2041-17232025-04-0116111210.1038/s41467-025-58705-6Highly dynamic and sensitive NEMOer calcium indicators for imaging ER calcium signals in excitable cellsWenjia Gu0Jia-Hui Chen1Yiyin Zhang2Zhirong Wang3Jia Li4Sijia Wang5Hanhan Zhang6Amin Jiang7Ziyi Zhong8Jiaxuan Zhang9Ze Xu10Panpan Liu11Chao Xi12Tingting Hou13Donald L. Gill14Dong Li15Yu Mu16Shi-Qiang Wang17Ai-Hui Tang18Youjun Wang19Beijing Key Laboratory of Gene Resource and Molecular Development, College of Life Sciences, Beijing Normal UniversityDepartment of Anatomy, School of Basic Medicine, Anhui Medical UniversityState Key Laboratory of Membrane Biology, College of Life Sciences, Peking UniversityInstitute of Neuroscience, State Key Laboratory of Neuroscience, Center for Excellence in Brain Science and Intelligence Technology, Chinese Academy of SciencesBeijing Key Laboratory of Gene Resource and Molecular Development, College of Life Sciences, Beijing Normal UniversityInstitute of Neuroscience, State Key Laboratory of Neuroscience, Center for Excellence in Brain Science and Intelligence Technology, Chinese Academy of SciencesBeijing Key Laboratory of Gene Resource and Molecular Development, College of Life Sciences, Beijing Normal UniversityNational Laboratory of Biomacromolecules, CAS Center for Excellence in Biomacromolecules, Institute of Biophysics, Chinese Academy of SciencesBeijing Key Laboratory of Gene Resource and Molecular Development, College of Life Sciences, Beijing Normal UniversityBeijing Key Laboratory of Gene Resource and Molecular Development, College of Life Sciences, Beijing Normal UniversityState Key Laboratory of Membrane Biology, College of Life Sciences, Peking UniversityBeijing Key Laboratory of Gene Resource and Molecular Development, College of Life Sciences, Beijing Normal UniversityBeijing Key Laboratory of Gene Resource and Molecular Development, College of Life Sciences, Beijing Normal UniversityState Key Laboratory of Membrane Biology, College of Life Sciences, Peking UniversityDepartment of Cellular and Molecular Physiology, Pennsylvania State University College of MedicineNational Laboratory of Biomacromolecules, CAS Center for Excellence in Biomacromolecules, Institute of Biophysics, Chinese Academy of SciencesInstitute of Neuroscience, State Key Laboratory of Neuroscience, Center for Excellence in Brain Science and Intelligence Technology, Chinese Academy of SciencesState Key Laboratory of Membrane Biology, College of Life Sciences, Peking UniversityAnhui Province Key Laboratory of Biomedical Imaging and Intelligent Processing, Institute of Artificial Intelligence, Hefei Comprehensive National Science CenterBeijing Key Laboratory of Gene Resource and Molecular Development, College of Life Sciences, Beijing Normal UniversityAbstract 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.https://doi.org/10.1038/s41467-025-58705-6
spellingShingle 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
Youjun Wang
Highly dynamic and sensitive NEMOer calcium indicators for imaging ER calcium signals in excitable cells
Nature Communications
title Highly dynamic and sensitive NEMOer calcium indicators for imaging ER calcium signals in excitable cells
title_full Highly dynamic and sensitive NEMOer calcium indicators for imaging ER calcium signals in excitable cells
title_fullStr Highly dynamic and sensitive NEMOer calcium indicators for imaging ER calcium signals in excitable cells
title_full_unstemmed Highly dynamic and sensitive NEMOer calcium indicators for imaging ER calcium signals in excitable cells
title_short Highly dynamic and sensitive NEMOer calcium indicators for imaging ER calcium signals in excitable cells
title_sort highly dynamic and sensitive nemoer calcium indicators for imaging er calcium signals in excitable cells
url https://doi.org/10.1038/s41467-025-58705-6
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