In Vivo and in Vitro Monitoring of Phagosome Maturation in Retinal Pigment Epithelium Cells

• DOI: 10.1007/978-1-4614-3209-8_11
• Publication Date: 2014-03-24T07:46:47Z
• Authors: Julian Esteve-Rudd; Vanda S. Lopes; Mei Jiang; David S. Williams
• Keywords: Mice; Opsins; Antibody Specificity; Phagosomes; Animals; Retinal Pigment Epithelium; In Vitro Techniques; Retinal Photoreceptor Cell Outer Segment; Cells, Cultured

The ingestion and degradation of photoreceptor disk membranes is a critical and major role for the retinal pigment epithelium (RPE). To help elucidate the cellular events involved in this role, functional in vivo and in vitro assays need to be developed further. Here we propose a method to help monitor phagosome maturation, using antibodies against different epitopes of opsin. We show that antibodies specific for the C-terminus of opsin label only immature phagosomes located in the apical region of the RPE. In contrast, antibodies recognizing the N-terminus also label more mature phagosomes, located more basally. The combined use of antibodies against different opsin epitopes thus provides a valuable tool in the study of phagosome maturation in the RPE.