Split genetically encoded calcium indicators for interorganellar junctions

• 成果类型: Article
• 署名作者: Olszakier, Shunit; Hussein, Wessal; Heinrich, Ronit; Andreyanov, Michael; Blau, Achinoam; Otor, Yara; Schiller, Jackie; Kellner, Shai; Berlin, Shai
• 署名单位: Technion Israel Institute of Technology; Rappaport Faculty of Medicine
• 刊物名称: PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
• ISSN/ISSBN: 0027-12223
• DOI: 10.1073/pnas.2415268122
• 发表日期: 2025-05-20
• 关键词: green fluorescent proteins; ca2+ indicators; mitochondria; translation; plasticity; dendrites; er; receptors; mechanism; neurons

摘要

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 Ca2+ 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.