Influence of zinc on selected cellular functions of cultured human retinal pigment epithelium.
Zinc is a necessary micronutrient, usually abundant in human RPE. Our study was undertaken to determine the effects of short-term, zinc deficiency on human retinal pigment epithelium (RPE) using a culture model of fetal human RPE cells. Human fetal RPE cells were isolated and cultured in Coon's modified Ham's F-12 medium. For zinc depletion studies, cells were cultured for 1 week in Chelex-treated Dulbecco's modified Eagle's medium containing low (0.25 microM) or physiologic (11 microM) total zinc concentrations as determined by flame atomic absorption spectroscopy. Protein synthesis was determined by incorporation of 35S-cysteine/methionine and labeled proteins analysed by polyacrylamide gel electrophoresis. Several cell parameters and enzymes were significantly reduced below control when cultured in low zinc: zinc content (40%), proliferation (63%), protein/well (50%), catalase activity (68%), alkaline phosphatase activity (61%), alpha-mannosidase activity (68%), and metallothionein (82%). No statistically significant decline was seen in acid phosphatase activity, superoxide dismutase activity, glutathione peroxidase activity and dexamethasone induction of metallothionein. Zinc repletion (100 microM, 1 h) increased catalase and alpha-mannosidase activities from 32% and 33% of control to 75% and 73%, respectively. Cycloheximide did not inhibit this short-term zinc-induced repletion of catalase or alpha-mannosidase. Protein synthesis in low zinc medium was depressed, but not significantly, as shown by incorporation of radiolabeled 35S-cysteine/methionine into newly synthesized proteins. The effects of zinc deficiency in cultured human RPE are selective. Adequate intracellular zinc was required for maximal activity of some enzymes. The dependence of catalase activity on zinc was not predicted and may help explain the observed decline in catalase activity seen with age in RPE. Our model of zinc deficiency should prove useful in elucidating the complex effects of zinc deficiency and repletion in human RPE.
Tate
D J
DJ
Sensory and Electrophysiology Research Unit, Touro Infirmary, New Orleans, LA 70115, USA.
Miceli
M V
MV
Newsome
D A
DA
Alcock
N W
NW
Oliver
P D
PD
D002455:Cell Division / Q000187:drug effects; D002478:Cells, Cultured; D002614:Chelating Agents / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D005136:Eye Proteins / Q000096:biosynthesis; D005333:Fetus / Q000166:cytology; D006801:Humans; D008361:Mannosidases / Q000378:metabolism; D008668:Metallothionein / Q000378:metabolism; D010088:Oxidoreductases / Q000378:metabolism; D010744:Phosphoric Monoester Hydrolases / Q000378:metabolism; D010857:Pigment Epithelium of Eye / Q000166:cytology / Q000187:drug effects / Q000196:embryology / Q000502:physiology*; D011137:Polystyrenes / Q000494:pharmacology; D011145:Polyvinyls / Q000494:pharmacology; D015032:Zinc / Q000172:deficiency / Q000494:pharmacology*; D043323:alpha-Mannosidase
D002614:Chelating Agents; D005136:Eye Proteins; D011137:Polystyrenes; D011145:Polyvinyls; C006960:chelex; D008668:Metallothionein; D010088:Oxidoreductases; D010744:Phosphoric Monoester Hydrolases; D008361:Mannosidases; D043323:alpha-Mannosidase; D015032:Zinc