epHero – a tandem-fluorescent probe to track the fate of apoptotic cells during efferocytosis

Author:

Singh SanjnaORCID,Bensalem JulienORCID,Hein Leanne K.,Casey Aaron,Mäkinen Ville-Petteri,Sargeant Timothy J.ORCID

Abstract

AbstractThe efficient removal of apoptotic cells via efferocytosis is critical for maintaining optimal tissue function. This involves the binding and engulfment of apoptotic cells by phagocytes and the subsequent maturation of the phagosome, culminating in lysosomal fusion and cargo destruction. However, current approaches to measure efferocytosis rely on labelling apoptotic targets with fluorescent dyes, which do not sufficiently distinguish between changes to the engulfment and acidification of apoptotic material. To address this limitation, we have developed a genetically coded ratiometric probe epHero which when expressed in the cytoplasm of target cells, bypasses the need for additional labelling steps. We demonstrate that epHero is a pH-sensitive reporter for efferocytosis and can be used to simultaneously track changes to apoptotic cell uptake and acidification, both in vitro and in mice. As proof-of-principle, we modify extracellular nutrition to show how epHero can distinguish between changes to cargo engulfment and acidification. Thus, tracking efferocytosis with epHero is a simple, cost-effective improvement on conventional techniques.

Funder

Australian Research Training Program Scholarship from the Australian Government

Publisher

Springer Science and Business Media LLC

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