Measuring the Acidification of the Phagosomal Lumen in Live C. elegans Embryos
Overview
Overview
Journal
STAR Protoc
Publisher
Cell Press
Specialties
Biology
Biomedical Engineering
Science
Biomedical Engineering
Science
Date
2023 Jun 4
PMID
37270784
Authors
Authors
Affiliations
Affiliations
Soon will be listed here.
Abstract
In metazoans, the acidification of the phagosomal lumen is essential for the efficient degradation of cargoes. Here, we present a protocol for measuring the rate of acidification inside phagosomal lumen containing apoptotic cells in living C. elegans embryos. We describe steps for generating a worm population, selecting embryos, and mounting embryos on agar pads. We then detail live imaging of embryos and data analysis. This protocol is applicable to any organism in which real-time fluorescence imaging can be performed. For complete details on the use and execution of this protocol, please refer to Pena-Ramos et al. (2022)..
References
1.
Treusch S, Knuth S, Slaugenhaupt S, Goldin E, Grant B, Fares H
. Caenorhabditis elegans functional orthologue of human protein h-mucolipin-1 is required for lysosome biogenesis. Proc Natl Acad Sci U S A. 2004; 101(13):4483-8.
PMC: 384773.
DOI: 10.1073/pnas.0400709101.
View
2.
Shen Q, He B, Lu N, Conradt B, Grant B, Zhou Z
. Phagocytic receptor signaling regulates clathrin and epsin-mediated cytoskeletal remodeling during apoptotic cell engulfment in C. elegans. Development. 2013; 140(15):3230-43.
PMC: 3931732.
DOI: 10.1242/dev.093732.
View
3.
Deretic V, Singh S, Master S, Harris J, Roberts E, Kyei G
. Mycobacterium tuberculosis inhibition of phagolysosome biogenesis and autophagy as a host defence mechanism. Cell Microbiol. 2006; 8(5):719-27.
DOI: 10.1111/j.1462-5822.2006.00705.x.
View
4.
Sokolovska A, Becker C, Stuart L
. Measurement of phagocytosis, phagosome acidification, and intracellular killing of Staphylococcus aureus. Curr Protoc Immunol. 2012; Chapter 14:14.30.1-14.30.12.
DOI: 10.1002/0471142735.im1430s99.
View
5.
Sulston J, Schierenberg E, White J, Thomson J
. The embryonic cell lineage of the nematode Caenorhabditis elegans. Dev Biol. 1983; 100(1):64-119.
DOI: 10.1016/0012-1606(83)90201-4.
View