Premium
Photophysical Studies of A2‐E, Putative Precursor of Lipofuscin, in Human Retinal Pigment Epithelial Cells
Author(s) -
Cubeddu Rinaldo,
Taroni Paola,
Hu DanNing,
Sakai Naomi,
Nakanishi Koji,
Roberts Joan E.
Publication year - 1999
Publication title -
photochemistry and photobiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.818
H-Index - 131
eISSN - 1751-1097
pISSN - 0031-8655
DOI - 10.1111/j.1751-1097.1999.tb07986.x
Subject(s) - lipofuscin , pigment , retinal , retinal pigment epithelium , fluorescence , biophysics , chemistry , retina , fluorescence microscope , biology , biochemistry , microbiology and biotechnology , photochemistry , optics , organic chemistry , physics , neuroscience
. With age, human retinal pigment epithelial cells accumulate lipofuscin that can absorb photons in the visible range leading to light‐induced damage and impaired vision. A putative precursor of lipofuscin, 2‐[2,6‐dimethyl‐8‐(2,6,6‐trimethyl‐l‐cyclohexen‐l‐yl)‐lE,3E:,5E,7E‐octa‐tetraenyl]‐l‐(2‐hydroxyethyl)‐4‐[4‐methyl‐6‐(2,6,6‐tri‐methyl‐1 ‐cyclohexen‐1 ‐y 1)‐ IE, 3E , 5E‐hexatrienyl] ‐pyridi‐nium (A2‐E), has recently been isolated and characterized from aged human retinal pigment epithelial cells. We have found that A2‐E inhibits the growth of human retinal pigment epithelial cells at concentrations greater than 1 μ M. Time‐resolved fluorescence measurements of 1 μ M A2‐E in solution, performed under 413 nm excitation, showed that fluorescence wave forms integrated across the spectrum (450–600 nm) were best‐fitted with three decay times in the nanosecond and subnanosecond time scale: 6.6,1.9 and 0.33 ns. Untreated retinal pigment epithelial cells were characterized by three fluorescence lifetimes: 6.3, 1.7 and 0.35 ns. In retinal pigment epithelial cells treated with 1 μ M A2‐E, the fluorescence decay was significantly faster, with the marked presence (30%) of a fourth short lifetime (0.12 ns). These fluorescence decay times for A2‐E bound to human retinal pigment epithelial cells are similar to those of lipofuscin granules isolated from aged human retinal pigment epithelial cells. This similarity supports the hypothesis that A2‐E is a precursor of lipofuscin and suggests that A2‐E may play a role in the overall light damage associated with age‐related retinal diseases.