Apoptotic Bodies: Selective Detection in Extracellular Vesicles

Abstract

Normal and dying cells release various types of membrane-bound vesicles including microvesicles, exosomes, and apoptotic bodies. These vesicles play important roles in intercellular communication and signal transduction. However, their diverse forms and subtypes fluctuate in size and other properties. In result current purification approaches do not fully discriminate between different categories of extracellular vesicles. Here, we present a fluorescence technique that specifically identifies apoptotic bodies in preparations of microvesicles, exosomes, and other extracellular vesicles.The approach exclusively labels the vesicles that contain DNA with 5'PO₄ blunt-ended DNA breaks, such as those produced by the apoptotic CAD nuclease during apoptotic DNA degradation. The technique can be useful in studies of apoptosis involving microvesicles and exosomes.

Keywords: 5′P blunt-ended DNA breaks; Apoptotic bodies; Apoptotic bodies labeling; Cell signaling in apoptosis; Clearance of apoptotic cells; Exosomes; Extracellular vesicles; In situ ligation technique; Microvesicles; “eat-me” signal; “find-me” signal.