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Ratiometric measurement of MAM Ca2+ dynamics using a modified CalfluxVTN

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  • Eunbyul Cho

    (Pohang University of Science and Technology)

  • Youngsik Woo

    (Pohang University of Science and Technology)

  • Yeongjun Suh

    (Pohang University of Science and Technology)

  • Bo Kyoung Suh

    (Pohang University of Science and Technology)

  • Soo Jeong Kim

    (Pohang University of Science and Technology)

  • Truong Thi My Nhung

    (Pohang University of Science and Technology)

  • Jin Yeong Yoo

    (Pohang University of Science and Technology)

  • Tran Diem Nghi

    (Pohang University of Science and Technology)

  • Su Been Lee

    (Pohang University of Science and Technology)

  • Dong Jin Mun

    (Pohang University of Science and Technology)

  • Sang Ki Park

    (Pohang University of Science and Technology)

Abstract

Mitochondria-associated ER membrane (MAM) is a structure where these calcium-regulating organelles form close physical contact sites for efficient Ca2+ crosstalk. Despite the central importance of MAM Ca2+ dynamics in diverse biological processes, directly and specifically measuring Ca2+ concentrations inside MAM is technically challenging. Here, we develop MAM-Calflux, a MAM-specific BRET-based Ca2+ indicator. The successful application of the bimolecular fluorescence complementation (BiFC) concept highlights Ca2+-responsive BRET signals in MAM. The BiFC strategy imparts dual functionality as a Ca2+ indicator and quantitative structural marker specific for MAM. As a ratiometric Ca2+ indicator, MAM-Calflux estimates steady-state MAM Ca2+ levels. Finally, it enables the visualization of uneven intracellular distribution of MAM Ca2+ and the elucidation of abnormally accumulated MAM Ca2+ from the neurons of Parkinson’s disease mouse model in both steady-state and stimulated conditions. Therefore, we propose that MAM-Calflux can be a versatile tool for ratiometrically measuring dynamic inter-organellar Ca2+ communication.

Suggested Citation

  • Eunbyul Cho & Youngsik Woo & Yeongjun Suh & Bo Kyoung Suh & Soo Jeong Kim & Truong Thi My Nhung & Jin Yeong Yoo & Tran Diem Nghi & Su Been Lee & Dong Jin Mun & Sang Ki Park, 2023. "Ratiometric measurement of MAM Ca2+ dynamics using a modified CalfluxVTN," Nature Communications, Nature, vol. 14(1), pages 1-14, December.
  • Handle: RePEc:nat:natcom:v:14:y:2023:i:1:d:10.1038_s41467-023-39343-2
    DOI: 10.1038/s41467-023-39343-2
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