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(Investigative Ophthalmology and Visual Science. 2004;45:2107-2114.)
© 2004 by The Association for Research in Vision and Ophthalmology, Inc.
DOI:  10.1167/iovs.03-1092
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Functional Properties of Hyalocytes under PDGF-Rich Conditions

Yoshihiro Noda,1 Yasuaki Hata,1 Toshio Hisatomi,1 Yuka Nakamura,1 Kumiko Hirayama,1 Muneki Miura,1 Shintaro Nakao,1 Kimihiko Fujisawa,1 Taiji Sakamoto,2 and Tatsuro Ishibashi1

1From the Department of Ophthalmology, Graduate School of Medical Sciences, Kyushu University, Fukuoka, Japan; and the 2Department of Ophthalmology, Faculty of Medicine, Kagoshima University, Kagoshima, Japan.

PURPOSE. To investigate the functional properties and intracellular signaling of hyalocytes under platelet-derived growth factor (PDGF)–rich conditions.

METHODS. The hyalocytes were isolated from bovine eyes and identified by immunocytochemistry and electron microscope. The expression of PDGF receptor {alpha}/ß and its phosphorylation in response to PDGF-BB was analyzed by Western blot analysis. PDGF-BB–induced proliferation and migration were evaluated by thymidine uptake and Boyden’s chemotaxis assay. The expression of the urokinase-type plasminogen activator (uPA) gene and the fibrinolytic activity were assessed by Northern blotting and fibrin zymography. An in vitro type I collagen gel contraction assay was performed to determine the effect of PDGF-BB on cellular contraction.

RESULTS. The hyalocytes were immunocytochemically positive for S-100 and negative for glial fibrillary acidic protein (GFAP) and cytokeratin, as previously described. The electron microscope demonstrated that hyalocytes possess lysosome-like granules, mitochondria, and micropinocytotic vesicles in their cytoplasm. The hyalocytes expressed PDGF receptor {alpha} and ß, both of which were immediately phosphorylated in response to PDGF-BB. PDGF-BB also activated p85 PI3-kinase, p44/p42 mitogen-activated protein (MAP) kinase and p38 MAP kinase. PDGF-BB induced thymidine uptake and migration in a concentration-dependent (0–10 ng/mL) manner. Inhibitors of the respective kinases prohibited PDGF-BB–dependent thymidine uptake and migration with the exception of the p44/p42 MAP kinase inhibitor, which displayed no inhibitory effects on migration. PDGF-BB increased uPA gene expression and fibrinolytic activity. Collagen gel contraction observed under PDGF-BB–rich conditions was not prohibited by the respective inhibitors investigated.

CONCLUSIONS. The hyalocytes demonstrated macrophage-like characteristics and may have both physiologic and pathologic roles, such as the maintenance of vitreous transparency through fibrinolytic activity and the pathogenesis of proliferative-vitreoretinal diseases through cellular proliferation and vitreous hyper-contraction.




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