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J Biol Chem, Vol. 275, Issue 18, 13918-13923, May 5, 2000
From the 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
Fibroblast Growth Factor-2 Promotes Keratan Sulfate Proteoglycan
Expression by Keratocytes in Vitro*
,,,,, and Division of Biology, Kansas State
University, Manhattan Kansas, 66506-4901 and the
§ Department of Ophthalmology, University of Pittsburgh
Medical School, Pittsburgh, Pennsylvania 15213-2588
, 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.
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