TY  - JOUR
AU  - Mauker, Philipp
AU  - Beckmann, Daniela
AU  - Kitowski, Annabel
AU  - Heise, Constanze
AU  - Wientjens, Chantal
AU  - Davidson, Andrew J
AU  - Wanderoy, Simone
AU  - Fabre, Gabin
AU  - Harbauer, Angelika B
AU  - Wood, Will
AU  - Wilhelm, Christoph
AU  - Thorn-Seshold, Julia
AU  - Misgeld, Thomas
AU  - Kerschensteiner, Martin
AU  - Thorn-Seshold, Oliver
TI  - Fluorogenic Chemical Probes for Wash-free Imaging of Cell Membrane Damage in Ferroptosis, Necrosis, and Axon Injury.
JO  - Journal of the American Chemical Society
VL  - 146
IS  - 16
SN  - 0002-7863
CY  - Washington, DC
PB  - ACS Publications
M1  - DZNE-2025-01275
SP  - 11072 - 11082
PY  - 2024
AB  - Selectively labeling cells with damaged membranes is needed not only for identifying dead cells in culture, but also for imaging membrane barrier dysfunction in pathologies in vivo. Most membrane permeability stains are permanently colored or fluorescent dyes that need washing to remove their non-uptaken extracellular background and reach good image contrast. Others are DNA-binding environment-dependent fluorophores, which lack design modularity, have potential toxicity, and can only detect permeabilization of cell volumes containing a nucleus (i.e., cannot delineate damaged volumes in vivo nor image non-nucleated cell types or compartments). Here, we develop modular fluorogenic probes that reveal the whole cytosolic volume of damaged cells, with near-zero background fluorescence so that no washing is needed. We identify a specific disulfonated fluorogenic probe type that only enters cells with damaged membranes, then is enzymatically activated and marks them. The esterase probe MDG1 is a reliable tool to reveal live cells that have been permeabilized by biological, biochemical, or physical membrane damage, and it can be used in multicolor microscopy. We confirm the modularity of this approach by also adapting it for improved hydrolytic stability, as the redox probe MDG2. We conclude by showing the unique performance of MDG probes in revealing axonal membrane damage (which DNA fluorogens cannot achieve) and in discriminating damage on a cell-by-cell basis in embryos in vivo. The MDG design thus provides powerful modular tools for wash-free in vivo imaging of membrane damage, and indicates how designs may be adapted for selective delivery of drug cargoes to these damaged cells: offering an outlook from selective diagnosis toward therapy of membrane-compromised cells in disease.
LB  - PUB:(DE-HGF)16
C6  - pmid:38592946
DO  - DOI:10.1021/jacs.3c07662
UR  - https://pub.dzne.de/record/282305
ER  -