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J Biol Chem, Vol. 275, Issue 18, 13918-13923, May 5, 2000

Fibroblast Growth Factor-2 Promotes Keratan Sulfate Proteoglycan Expression by Keratocytes in Vitro*

Chad J. LongDagger , Mary R. RothDagger , Elena S. TashevaDagger , Martha Funderburgh§, Rachel SmitDagger , Gary W. ConradDagger , and James L. Funderburgh§

From the Dagger  Division of Biology, Kansas State University, Manhattan Kansas, 66506-4901 and the § Department of Ophthalmology, University of Pittsburgh Medical School, Pittsburgh, Pennsylvania 15213-2588

Keratocytes of the corneal stroma produce a specialized extracellular matrix responsible for corneal transparency. Corneal keratan sulfate proteoglycans (KSPG) are unique products of keratocytes that are down-regulated in corneal wounds and in vitro. This study used cultures of primary bovine keratocytes to define factors affecting KSPG expression in vitro. KSPG metabolically labeled with [35S]sulfate decreased during the initial 2-4 days of culture in quiescent cultures with low serum concentrations (0.1%). Addition of fetal bovine serum, fibroblast growth factor-2 (FGF-2), transforming growth factor beta , or platelet derived growth factor all stimulated cell division, but only FGF-2 stimulated KSPG secretion. Combined with serum, FGF-2 also prevented serum-induced KSPG down-regulation. KSPG secretion was lost during serial subculture with or without FGF-2. Expression of KSPG core proteins (lumican, mimecan, and keratocan) was stimulated by FGF-2, and steady state mRNA pools for these proteins, particularly keratocan, were significantly increased by FGF-2 treatment. KSPG expression therefore is supported by exogenous FGF-2 and eliminated by subculture of the cells in presence of serum. FGF-2 stimulates KSPG core protein expression primarily through an increase in mRNA pools.

* This work was supported by National Institutes of Health Grants EY09368 (to J. L. F.) EY08098 (Core grant to the University of Pittsburgh, Ophthalmology), and EY00952 (to G. W. C.), by NASA Grant NASA-NAGW 2368 (to G. W. C.), and by funds from Research to Prevent Blindness and the Eye and Ear Foundation of Pittsburgh.The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

Jules and Doris Stein Research to Prevent Blindness Professor. To whom correspondence should be addressed: Dept. of Ophthalmology, University of Pittsburgh, 203 Lothrop St., Pittsburgh, PA 15213. Tel.: 412-647-3853; Fax: 412-647-5880; E-mail: jlfunder@pitt.edu.

Copyright © 2000 by The American Society for Biochemistry and Molecular Biology, Inc.
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