Split genetically encoded calcium indicators for interorganellar junctions
Abstract
Genetically encoded calcium indicators (GECIs) have revolutionized the study of cellular calcium signaling, offering powerful tools for real-time optical monitoring of calcium dynamics. Although contemporary GECIs can be targeted to various organelles, there are no means to obtain active and functional GECIs exclusively at interorganellar junctions. To address this gap, we have developed a toolbox of split versions of green and red GECIs designed to reassemble only when the two “halves” come into proximity. We developed split probes to investigate interorganellar connectivity and activity between mitochondria and the ER (via split-MEGIC) or between the plasma membrane and the ER (via split-sf-MEMBER). We employ the various split-sensors to image neural Ca 2+ activity in vitro and in vivo and, in the process, identify Mito–ER junctions and calcium activity within individual dendritic spines by use of split-MEGIC.
Article Details
Journal Info
Proceedings of the National Academy of Sciences
National Academy of Sciences
Authors (9)
Shunit Olszakier
Department of Neuroscience, Ruth and Bruce Rappaport Faculty of Medicine, Technion–Israel Institute of Technology
Wessal Hussein
Department of Neuroscience, Ruth and Bruce Rappaport Faculty of Medicine, Technion–Israel Institute of Technology
Ronit Heinrich
Department of Neuroscience, Ruth and Bruce Rappaport Faculty of Medicine, Technion–Israel Institute of Technology
Michael Andreyanov
Department of Neuroscience, Ruth and Bruce Rappaport Faculty of Medicine, Technion–Israel Institute of Technology
Achinoam Blau
Department of Neuroscience, Ruth and Bruce Rappaport Faculty of Medicine, Technion–Israel Institute of Technology
Yara Otor
Department of Neuroscience, Ruth and Bruce Rappaport Faculty of Medicine, Technion–Israel Institute of Technology
Jackie Schiller
Department of Neuroscience, Ruth and Bruce Rappaport Faculty of Medicine, Technion–Israel Institute of Technology
Shai Kellner
Department of Neuroscience, Ruth and Bruce Rappaport Faculty of Medicine, Technion–Israel Institute of Technology
Shai Berlin
Department of Neuroscience, Ruth and Bruce Rappaport Faculty of Medicine, Technion–Israel Institute of Technology