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Changes in retinal pigment epithelial cell autofluorescence and protein expression associated with phagocytosis of rod outer segments in vitro
Author(s) -
Rakoczy Piroska,
Kennedy Christopher,
ThompsonWallis Dawn,
Mann Krishna,
Constable Ian
Publication year - 1992
Publication title -
biology of the cell
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.543
H-Index - 85
eISSN - 1768-322X
pISSN - 0248-4900
DOI - 10.1016/0248-4900(92)90194-6
Subject(s) - autofluorescence , biology , phagocytosis , lipofuscin , flow cytometry , retinal , retinal pigment epithelium , in vitro , retina , pigment , microbiology and biotechnology , cell culture , biochemistry , fluorescence , chemistry , genetics , physics , organic chemistry , quantum mechanics , neuroscience
Summary— The accumulation of autofluorescent lipofuscin was quantified in cultured human retinal pigment epithelial (RPE) cells phagocytosing bovine rod outer segments (BROS) and the expression of proteins in these cells was investigated. Results showed a steady increase in autofluorescence of RPE cells over a 4‐week period as measured by fluorophotometric flow cytometry. A significantly greater increase in autofluorescence was found in the cultured RPE cells from a 7‐year‐old donor compared with those from a 47‐year‐old donor. Within both groups the BROs‐challenged cells had significantly higher fluorescence readings than the control cells which were not challenged. Autoradiography of 35 S‐labelled proteins separated by polyacrylamide gel electrophoresis (PAGE) revealed a small distinct band at 102 kDa in BROS‐challenged RPE cells of both bovine and human origin that did not appear in control or microsphere‐phagocytosing RPE cells. The intensity of the signal was unrelated to the duration of the challenge period.