Biological roles for FGF-5 in choroidal neovascularization

M. A Z Keithahn, A. E. Aotaki-Keen, M. Friedlander, Leonard M Hjelmeland, Lawrence S Morse

Research output: Contribution to journalArticle

Abstract

Purpose. Previously we have shown that bovine choroidal microvascular endothelial cells exhibit in vitro expression of FGF-5 at the mRNA and protein levels. The purpose of this study was to investigate the possible biological roles of FGF-5 for cell types involved in the process of choroidal neovascularization. Methods. The mitogenic activity of commercially obtained human recombinant FGF-5 was examined using a cell counting-based proliferation assay. A variety of cell types including bovine choroidal microvascular endothelial, fetal bovine heart endothelial, bovine aortic endothelial, human umbilical vein endothelial, Balb/c3T3 fibroblasts, and human retinal pigment epithelial (RPE) cells were tested. Balb/c3T3 cells were used as a cell line which is known to respond mitogenically to the commercial preparation of FGF-5. Basic FGF served as a positive mitogenic control for all cell lines. Results. Balb/c3T3 fibroblasts exhibited a mitogenic response to FGF-5 with an approximate ED50 of 1.8-3.7 nM which is within the range reported by the manufacturer using a similar embryonic fibroblast cell line. FGF-5 was also found to stimulate proliferation of RPE cells. Basic FGF displayed mitogenic activity for all cell lines tested with ED50's ranging from 2-100 pM. Human recombinant FGF-5 was not mitogenic for any of the endothelial cell lines tested. Conclusions. Human recombinant FGF-5 does not appear to be a mitogen for endothelial cells in vitro. This commercially prepared FGF-5 lacks carbohydrate modification found in the protein produced by eukaryotic cells. Our current studies are focused on the role which glycosylation may play in stabilizing FGF-5 and thus the biological activity of this protein. We are also investigating the possibility that FGF-5 is chemotactic in vitro without being mitogenic for these cells. FGF-5 could also be serving a strictly paracrine role in the stimulation of fibroblasts and/or RPE cells during the formation of choroidal neovascular membranes.

Original languageEnglish (US)
JournalInvestigative Ophthalmology and Visual Science
Volume37
Issue number3
StatePublished - Feb 15 1996

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Choroidal Neovascularization
Retinal Pigments
Cell Line
Fibroblasts
Endothelial Cells
Epithelial Cells
Fetal Heart
Umbilical Veins
Proteins
Eukaryotic Cells
Glycosylation
Mitogens
Carbohydrates
Messenger RNA
Membranes
In Vitro Techniques

ASJC Scopus subject areas

  • Ophthalmology

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Biological roles for FGF-5 in choroidal neovascularization. / Keithahn, M. A Z; Aotaki-Keen, A. E.; Friedlander, M.; Hjelmeland, Leonard M; Morse, Lawrence S.

In: Investigative Ophthalmology and Visual Science, Vol. 37, No. 3, 15.02.1996.

Research output: Contribution to journalArticle

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abstract = "Purpose. Previously we have shown that bovine choroidal microvascular endothelial cells exhibit in vitro expression of FGF-5 at the mRNA and protein levels. The purpose of this study was to investigate the possible biological roles of FGF-5 for cell types involved in the process of choroidal neovascularization. Methods. The mitogenic activity of commercially obtained human recombinant FGF-5 was examined using a cell counting-based proliferation assay. A variety of cell types including bovine choroidal microvascular endothelial, fetal bovine heart endothelial, bovine aortic endothelial, human umbilical vein endothelial, Balb/c3T3 fibroblasts, and human retinal pigment epithelial (RPE) cells were tested. Balb/c3T3 cells were used as a cell line which is known to respond mitogenically to the commercial preparation of FGF-5. Basic FGF served as a positive mitogenic control for all cell lines. Results. Balb/c3T3 fibroblasts exhibited a mitogenic response to FGF-5 with an approximate ED50 of 1.8-3.7 nM which is within the range reported by the manufacturer using a similar embryonic fibroblast cell line. FGF-5 was also found to stimulate proliferation of RPE cells. Basic FGF displayed mitogenic activity for all cell lines tested with ED50's ranging from 2-100 pM. Human recombinant FGF-5 was not mitogenic for any of the endothelial cell lines tested. Conclusions. Human recombinant FGF-5 does not appear to be a mitogen for endothelial cells in vitro. This commercially prepared FGF-5 lacks carbohydrate modification found in the protein produced by eukaryotic cells. Our current studies are focused on the role which glycosylation may play in stabilizing FGF-5 and thus the biological activity of this protein. We are also investigating the possibility that FGF-5 is chemotactic in vitro without being mitogenic for these cells. FGF-5 could also be serving a strictly paracrine role in the stimulation of fibroblasts and/or RPE cells during the formation of choroidal neovascular membranes.",
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