PUBLICATION
SPLICS: a split green fluorescent protein-based contact site sensor for narrow and wide heterotypic organelle juxtaposition
- Authors
- Cieri, D., Vicario, M., Giacomello, M., Vallese, F., Filadi, R., Wagner, T., Pozzan, T., Pizzo, P., Scorrano, L., Brini, M., Calì, T.
- ID
- ZDB-PUB-190917-7
- Date
- 2018
- Source
- Cell death and differentiation 25: 1131-1145 (Journal)
- Registered Authors
- Keywords
- none
- MeSH Terms
-
- HEK293 Cells
- Mitochondrial Membranes/metabolism*
- Endoplasmic Reticulum/genetics
- Endoplasmic Reticulum/metabolism*
- Calcium Signaling*
- Autophagy*
- Apoptosis*
- Green Fluorescent Proteins/genetics
- Green Fluorescent Proteins/metabolism
- HeLa Cells
- Humans
- Mitochondria/genetics
- Mitochondria/metabolism*
- Animals
- Zebrafish
- PubMed
- 29229997 Full text @ Cell Death Differ.
Citation
Cieri, D., Vicario, M., Giacomello, M., Vallese, F., Filadi, R., Wagner, T., Pozzan, T., Pizzo, P., Scorrano, L., Brini, M., Calì, T. (2018) SPLICS: a split green fluorescent protein-based contact site sensor for narrow and wide heterotypic organelle juxtaposition. Cell death and differentiation. 25:1131-1145.
Abstract
Contact sites are discrete areas of organelle proximity that coordinate essential physiological processes across membranes, including Ca2+ signaling, lipid biosynthesis, apoptosis, and autophagy. However, tools to easily image inter-organelle proximity over a range of distances in living cells and in vivo are lacking. Here we report a split-GFP-based contact site sensor (SPLICS) engineered to fluoresce when organelles are in proximity. Two SPLICS versions efficiently measured narrow (8-10 nm) and wide (40-50 nm) juxtapositions between endoplasmic reticulum and mitochondria, documenting the existence of at least two types of contact sites in human cells. Narrow and wide ER-mitochondria contact sites responded differently to starvation, ER stress, mitochondrial shape modifications, and changes in the levels of modulators of ER-mitochondria juxtaposition. SPLICS detected contact sites in soma and axons of D. rerio Rohon Beard (RB) sensory neurons in vivo, extending its use to analyses of organelle juxtaposition in the whole animal.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping