Lysosomes contain hydrolytic enzymes that can degrade proteins and DNA. Leakage of these reactive compounds through a compromised lysosomal membrane causes lysosomal cell death, which can have apoptotic, necrotic, or mixed morphology. Lysosomal cathepsin proteases, such as cathepsin D, and the lysosomal endonuclease, DNase II, have both been implicated in lysosome-related cell death. Here, we present a fluorescence dual-labeling technique for simultaneous visualization of these two markers of lysosomal activity linked to cell death. The approach labels the intracellular distribution of cathepsin D and the sites with DNase II-type breaks in fixed tissue sections. It determines the lysosomal or extra-lysosomal localization of the markers and can be useful in studying pathways and signals of lysosomal cell death.
Keywords: Cathepsin D; Clearance of apoptotic cells; DNA breaks; DNase II; DNase II-type breaks; Florescence dual-labeling technique; Lysosomal cell death; Lysosomes; Phagolysosomes; Vaccinia topoisomerase-based labeling.