Ratiometric Measurement of MAM Ca Dynamics Using a Modified CalfluxVTN

Overview

Journal Nat Commun

Specialty Biology

Date 2023 Jun 16

PMID 37328454

Authors

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

Affiliations

Soon will be listed here.

Abstract

Mitochondria-associated ER membrane (MAM) is a structure where these calcium-regulating organelles form close physical contact sites for efficient Ca crosstalk. Despite the central importance of MAM Ca dynamics in diverse biological processes, directly and specifically measuring Ca concentrations inside MAM is technically challenging. Here, we develop MAM-Calflux, a MAM-specific BRET-based Ca indicator. The successful application of the bimolecular fluorescence complementation (BiFC) concept highlights Ca-responsive BRET signals in MAM. The BiFC strategy imparts dual functionality as a Ca indicator and quantitative structural marker specific for MAM. As a ratiometric Ca indicator, MAM-Calflux estimates steady-state MAM Ca levels. Finally, it enables the visualization of uneven intracellular distribution of MAM Ca and the elucidation of abnormally accumulated MAM Ca 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 Ca communication.

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