Ratiometric measurement of MAM Ca2+ dynamics using a modified CalfluxVTN
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+ co...
| Main Authors: | , , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Nature Portfolio
2023-06-01
|
| Series: | Nature Communications |
| Online Access: | https://doi.org/10.1038/s41467-023-39343-2 |
| _version_ | 1827922836491075584 |
|---|---|
| author | 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 |
| author_facet | 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 |
| author_sort | Eunbyul Cho |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-03-13T04:48:26Z |
| format | Article |
| id | doaj.art-6a6f8fa77b9946719b3ea11aa09bf31e |
| institution | Directory Open Access Journal |
| issn | 2041-1723 |
| language | English |
| last_indexed | 2024-03-13T04:48:26Z |
| publishDate | 2023-06-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Nature Communications |
| spelling | doaj.art-6a6f8fa77b9946719b3ea11aa09bf31e2023-06-18T11:19:22ZengNature PortfolioNature Communications2041-17232023-06-0114111410.1038/s41467-023-39343-2Ratiometric measurement of MAM Ca2+ dynamics using a modified CalfluxVTNEunbyul Cho0Youngsik Woo1Yeongjun Suh2Bo Kyoung Suh3Soo Jeong Kim4Truong Thi My Nhung5Jin Yeong Yoo6Tran Diem Nghi7Su Been Lee8Dong Jin Mun9Sang Ki Park10Department of Life Sciences, Pohang University of Science and TechnologyDepartment of Life Sciences, Pohang University of Science and TechnologyDepartment of Life Sciences, Pohang University of Science and TechnologyDepartment of Life Sciences, Pohang University of Science and TechnologyDepartment of Life Sciences, Pohang University of Science and TechnologyDepartment of Life Sciences, Pohang University of Science and TechnologyDepartment of Life Sciences, Pohang University of Science and TechnologyDepartment of Life Sciences, Pohang University of Science and TechnologyDepartment of Life Sciences, Pohang University of Science and TechnologyDepartment of Life Sciences, Pohang University of Science and TechnologyDepartment of Life Sciences, Pohang University of Science and TechnologyAbstract 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.https://doi.org/10.1038/s41467-023-39343-2 |
| spellingShingle | 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 Ratiometric measurement of MAM Ca2+ dynamics using a modified CalfluxVTN Nature Communications |
| title | Ratiometric measurement of MAM Ca2+ dynamics using a modified CalfluxVTN |
| title_full | Ratiometric measurement of MAM Ca2+ dynamics using a modified CalfluxVTN |
| title_fullStr | Ratiometric measurement of MAM Ca2+ dynamics using a modified CalfluxVTN |
| title_full_unstemmed | Ratiometric measurement of MAM Ca2+ dynamics using a modified CalfluxVTN |
| title_short | Ratiometric measurement of MAM Ca2+ dynamics using a modified CalfluxVTN |
| title_sort | ratiometric measurement of mam ca2 dynamics using a modified calfluxvtn |
| url | https://doi.org/10.1038/s41467-023-39343-2 |
| work_keys_str_mv | AT eunbyulcho ratiometricmeasurementofmamca2dynamicsusingamodifiedcalfluxvtn AT youngsikwoo ratiometricmeasurementofmamca2dynamicsusingamodifiedcalfluxvtn AT yeongjunsuh ratiometricmeasurementofmamca2dynamicsusingamodifiedcalfluxvtn AT bokyoungsuh ratiometricmeasurementofmamca2dynamicsusingamodifiedcalfluxvtn AT soojeongkim ratiometricmeasurementofmamca2dynamicsusingamodifiedcalfluxvtn AT truongthimynhung ratiometricmeasurementofmamca2dynamicsusingamodifiedcalfluxvtn AT jinyeongyoo ratiometricmeasurementofmamca2dynamicsusingamodifiedcalfluxvtn AT trandiemnghi ratiometricmeasurementofmamca2dynamicsusingamodifiedcalfluxvtn AT subeenlee ratiometricmeasurementofmamca2dynamicsusingamodifiedcalfluxvtn AT dongjinmun ratiometricmeasurementofmamca2dynamicsusingamodifiedcalfluxvtn AT sangkipark ratiometricmeasurementofmamca2dynamicsusingamodifiedcalfluxvtn |