Imaging Interorganelle Contacts and Local Calcium Dynamics at the ER-mitochondrial Interface

Overview

Journal Mol Cell

Publisher Cell Press

Specialty Cell Biology

Date 2010 Jul 7

PMID 20603080

Citations 360

Authors

Gyorgy Csordas

Peter Varnai

Tunde Golenar

Swati Roy

George Purkins

Timothy G Schneider

Tamas Balla

Gyorgy Hajnoczky

Affiliations

Soon will be listed here.

Abstract

The ER-mitochondrial junction provides a local calcium signaling domain that is critical for both matching energy production with demand and the control of apoptosis. Here, we visualize ER-mitochondrial contact sites and monitor the localized [Ca(2+)] changes ([Ca(2+)](ER-mt)) using drug-inducible fluorescent interorganelle linkers. We show that all mitochondria have contacts with the ER, but plasma membrane (PM)-mitochondrial contacts are less frequent because of interleaving ER stacks in both RBL-2H3 and H9c2 cells. Single mitochondria display discrete patches of ER contacts and show heterogeneity in the ER-mitochondrial Ca(2+) transfer. Pericam-tagged linkers revealed IP(3)-induced [Ca(2+)](ER-mt) signals that exceeded 9 microM and endured buffering bulk cytoplasmic [Ca(2+)] increases. Altering linker length to modify the space available for the Ca(2+) transfer machinery had a biphasic effect on [Ca(2+)](ER-mt) signals. These studies provide direct evidence for the existence of high-Ca(2+) microdomains between the ER and mitochondria and suggest an optimal gap width for efficient Ca(2+) transfer.

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