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Expression of neurotrophic factors in cultured human retinal pigment epithelial cells
K Ishida1, N Yoshimura, M Yoshida
1Department of Ophthalmology and Visual Science, Kyoto University Graduate School of Medicine, Japan.
Purpose:
To see if cultured human retinal pigment epithelial (RPE) cells have the capacity to synthesize neurotrophins, including nerve growth factor (NGF), brain-derived growth factor (BDNF), and neurotrophin-3 (NT-3).
Methods:
Expression of mRNAs for the neurotrophins was studied by the reverse transcription polymerase chain reaction (PCR) method. Quantitative analysis of the gene expression was done by using a semiquantitative PCR method. Secretion of NGF-like immunoreactivity (NGF-LI) into the culture medium was analyzed by enzyme immunoassay (EIA).
Results:
Cultured human RPE cells were found to express mRNAs for NGF, BDNF and NT-3. In the conditioned culture medium of the human RPE, 9.44 +/- 0.62 pg/ml (mean +/- SEM, n = 6) NGF-LI was found. Pretreatment of human RPE cells with interleukin-l (IL-1) (20 ng/ml), phorbol myristate acetate (PMA) (100 ng/ml) or tumor necrosis factor-alpha (TNF-alpha) (40 ng/ml) was found to increase the mRNA expression of neurotrophins and also to increase secretion of NGF-LI into the culture medium.
Conclusions:
Our data demonstrate that cultured human RPE cells have the capacity to synthesize neurotrophins, and that various stimulations can up-regulate gene and protein expression of NGF by these cells.