title: Heterogeneity in ornithine cytotoxicity of bovine retinal pigment epithelial cells in primary culture.
abstract: Gyrate atrophy of the choroid and retina is a chorioretinal degeneration caused by hyperornithinemia and a deficiency of ornithine-delta-aminotransferase (OAT). We recently showed that ornithine exhibits cytotoxicity to human retinal pigment epithelial (RPE) cell lines treated with the OAT inhibitor, 5-fluoromethylornithine (5-FMOrn), and suggested that this system may be an in vitro model of gyrate atrophy. In the present study, in order to apply this system to primary cultured RPE cells, we freshly prepared RPE cells from bovine eyes and studied the effect of ornithine on cell damage. Two phenotypes, epithelioid and fusiform, which coexisted in the primary culture and epithelioid phenotype cells, but not fusiform ones, were severely damaged and partially detached from the substrate by 10 m m ornithine and 0.5 m m 5-FMOrn. Neither ornithine nor 5-FMOrn alone exhibited such cytotoxicity to both phenotypes of RPE cells. Proline significantly prevented the ornithine-induced cytotoxicity. Epithelioid and fusiform phenotypes isolated from the primary culture showed different distribution of actin filaments. A combination of ornithine and 5-FMOrn time-dependently inhibited [(3)H]thymidine incorporation in the epithelioid, but not fusiform, cells. Proline prevented the inhibition of [(3)H]thymidine incorporation by ornithine in 5-FMOrn-treated epithelioid cells. Furthermore, l -azetidine-2-carboxylic acid, a collagen synthesis inhibitor, reduced [(3)H]thymidine incorporation in epithelioid, but not fusiform, cells, which was reversed by proline. These results demonstrate that the epithelioid phenotype of bovine RPE cells becomes susceptible to ornithine following inactivation of OAT. The phenotypic cells and its prevention by proline may provide insight into biochemical triggers that induce gyrate atrophy.
authors.affiliation: Department of Ophthalmology, Kansai Medical University, 10-15 Fumizono, Moriguchi, Osaka, 570-8506, Japan.
authors.forename: A
authors.initials: A
authors.lastname: Ando
authors.forename: M
authors.initials: M
authors.lastname: Ueda
authors.forename: M
authors.initials: M
authors.lastname: Uyama
authors.forename: Y
authors.initials: Y
authors.lastname: Masu
authors.forename: T
authors.initials: T
authors.lastname: Okumura
authors.forename: S
authors.initials: S
authors.lastname: Ito
mesh_terms: D000818:Animals; D002417:Cattle; D016923:Cell Death; D002478:Cells, Cultured; D004247:DNA / Q000096:biosynthesis; D004357:Drug Synergism; D004791:Enzyme Inhibitors / Q000494:pharmacology; D015799:Gyrate Atrophy / Q000378:metabolism*; D009952:Ornithine / Q000009:adverse effects* / Q000031:analogs & derivatives / Q000494:pharmacology; D009953:Ornithine-Oxo-Acid Transaminase / Q000037:antagonists & inhibitors; D010857:Pigment Epithelium of Eye / Q000166:cytology / Q000187:drug effects* / Q000201:enzymology; D011392:Proline / Q000494:pharmacology
chemical_list: D004791:Enzyme Inhibitors; C057405:5-fluoromethylornithine; D004247:DNA; D011392:Proline; D009952:Ornithine; D009953:Ornithine-Oxo-Acid Transaminase