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<p>The Endoplasmic/sarcoplasmic reticulum (ER/SR) is central to calcium (Ca<sup>2+</sup>) signaling, yet current genetically encoded Ca<sup>2+</sup> indicators (GECIs) cannot detect elementary Ca<sup>2+</sup> release events from ER/SR, particularly in muscle cells. Here, we report NEMOer, a set of organellar GECIs, to efficiently capture ER Ca<sup>2+</sup> 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 Ca<sup>2+</sup> transients in both non-excitable and excitable cells. The ratiometric version further allows super-resolution monitoring of local ER Ca<sup>2+</sup> homeostasis and dynamics. Notably, NEMOer-f enabled the inaugural detection of Ca<sup>2+</sup> blinks, elementary Ca<sup>2+</sup> releasing signals from the SR of cardiomyocytes, as well as in vivo spontaneous SR Ca<sup>2+</sup> releases in zebrafish. In summary, the highly dynamic NEMOer sensors expand the repertoire of organellar Ca<sup>2+</sup> sensors that allow real-time monitoring of intricate Ca<sup>2+</sup> dynamics and homeostasis in live cells with high spatiotemporal resolution.</p>

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

  • 期刊论文

  • 出版模式：

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  • 发表日期：

    2025年4月11日

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  • 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

摘要

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.