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  • Title: Modulation of early response gene expression by prostaglandins in cultured rat retinal pigment epithelium cells.
    Author: Ershov AV, Parkins N, Lukiw WJ, Bazan NG.
    Journal: Curr Eye Res; 2000 Dec; 21(6):968-74. PubMed ID: 11262621.
    Abstract:
    PURPOSE: To explore the role of prostaglandins (PGs) as modulators of retinal pigment epithelium (RPE) rod outer segment (ROS)-phagocytosis and ROS-phagocytosis-induced gene expression. METHODS: RPE cells in primary cell culture were pre-incubated with PGE( 2), PGD(2), PGF(2)alpha, PGJ(2), 15-deoxy-Delta( 12,14)-PGJ(2) or U-46619 (stable analog of thromboxane A(2)), and fed with a suspension of ROS. Expression of zif-268 and tis-1 mRNA was determined by Northern blotting. DNA-binding activity of TIS-1 protein was assessed by electrophoretic mobility shift assay. Concentration of PGE (2) and PGD (2) in the tissue culture medium was measured by enzyme immuno-assay. Phagocytis-tosis was quantified by counting of double-immunostained bound and ingested ROS. RESULTS: PGE 2, the most potent of PGs, strongly elevated both basal and ROS-phagocytosis-induced levels of tis-1 mRNA, while significantly inhibiting both basal and phagocytosis-induced expression of zif-268 mRNA. PGD(2), PGJ(2) and 15-deoxy-Delta(12,14)-PGJ( 2) elevated ROS-phagocytosis-induced, but not basal, expression of tis-1 mRNA expression. PGF(2alpha) super-induced both phagocytosis-induced and basal tis-1 mRNA expression. U-46619 and carbaprostacyclin had no effect on expression of tis-1 mRNA. PGE(2) was the only PG to affect zif-268 expression. Exogenous PGE(2), PGD( 2) and PGF(2alpha), when added to the medium at 1-microM concentrations, significantly inhibited ingestion of ROS, with PGE(2) being the most potent PG affecting ROS-phagocytosis. CONCLUSIONS: PGs act as selective regulators of phagocytosis-induced transcription factor gene expression in RPE cells, as well as of ROS-phagocytosis itself. This modulation may help to ensure specificity in the differential activation of target genes by ROS-phagocytosis receptor-mediated signal transduction in RPE cells.
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