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Improved calcium sensor GCaMP-X overcomes the calcium channel perturbations induced by the calmodulin in GCaMP

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  • Yaxiong Yang

    (Tsinghua University
    Beihang University
    Beihang University
    Tsinghua University)

  • Nan Liu

    (Tsinghua University
    Yunan University)

  • Yuanyuan He

    (Tsinghua University)

  • Yuxia Liu

    (Tsinghua University)

  • Lin Ge

    (Tsinghua University)

  • Linzhi Zou

    (Tsinghua University)

  • Sen Song

    (Tsinghua University
    Tsinghua University)

  • Wei Xiong

    (Tsinghua University
    Tsinghua University)

  • Xiaodong Liu

    (Tsinghua University
    Beihang University
    Beihang University
    Tsinghua University)

Abstract

GCaMP, one popular type of genetically-encoded Ca2+ indicator, has been associated with various side-effects. Here we unveil the intrinsic problem prevailing over different versions and applications, showing that GCaMP containing CaM (calmodulin) interferes with both gating and signaling of L-type calcium channels (CaV1). GCaMP acts as an impaired apoCaM and Ca2+/CaM, both critical to CaV1, which disrupts Ca2+ dynamics and gene expression. We then design and implement GCaMP-X, by incorporating an extra apoCaM-binding motif, effectively protecting CaV1-dependent excitation–transcription coupling from perturbations. GCaMP-X resolves the problems of detrimental nuclear accumulation, acute and chronic Ca2+ dysregulation, and aberrant transcription signaling and cell morphogenesis, while still demonstrating excellent Ca2+-sensing characteristics partly inherited from GCaMP. In summary, CaM/CaV1 gating and signaling mechanisms are elucidated for GCaMP side-effects, while allowing the development of GCaMP-X to appropriately monitor cytosolic, submembrane or nuclear Ca2+, which is also expected to guide the future design of CaM-based molecular tools.

Suggested Citation

  • Yaxiong Yang & Nan Liu & Yuanyuan He & Yuxia Liu & Lin Ge & Linzhi Zou & Sen Song & Wei Xiong & Xiaodong Liu, 2018. "Improved calcium sensor GCaMP-X overcomes the calcium channel perturbations induced by the calmodulin in GCaMP," Nature Communications, Nature, vol. 9(1), pages 1-18, December.
    • Handle: RePEc:nat:natcom:v:9:y:2018:i:1:d:10.1038_s41467-018-03719-6
    DOI: 10.1038/s41467-018-03719-6
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    Cited by:

    1. Han Chin Wang & Amy M. LeMessurier & Daniel E. Feldman, 2022. "Tuning instability of non-columnar neurons in the salt-and-pepper whisker map in somatosensory cortex," Nature Communications, Nature, vol. 13(1), pages 1-18, December.
    2. Sverre Grødem & Ingeborg Nymoen & Guro Helén Vatne & Frederik Sebastian Rogge & Valgerður Björnsdóttir & Kristian Kinden Lensjø & Marianne Fyhn, 2023. "An updated suite of viral vectors for in vivo calcium imaging using intracerebral and retro-orbital injections in male mice," Nature Communications, Nature, vol. 14(1), pages 1-13, December.

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