Funaki H, Sawaguchi S, Yaoeda K, Koyama Y, Yaoita E, Funaki S, Shirakashi M, Oshima Y, Shukunami C, Hiraki Y, Abe H, Yamamoto T
Department of Structural Pathology, Institute of Nephrology, Niigata University, 1-757 Asahimachi-dori, Niigata 951-8510, Japan.
Invest Ophthalmol Vis Sci. 2001 May;42(6):1193-200.
To determine the role in the eye of chondromodulin (ChM)-I, which has been identified in cartilage as an angiogenic inhibitor, the expression and localization and a possible function of ChM-I were investigated.
Expression and localization of ChM-I in rat eyes were examined by RNase protection assay and in situ hybridization and by immunostaining, using an antibody against a synthetic peptide. The effect of recombinant ChM-I on tube morphogenesis of retinal endothelial cells was examined in culture.
The rat ChM-I gene was determined to encode the open reading frame of 334 amino acid residues, and ChM-I mRNA was exclusively expressed in cartilage, eye, and cerebellum in rats. ChM-I mRNA expression was evident in the iris-ciliary body, retina, and scleral compartments, but not in other compartments of the eye. In situ hybridization revealed mRNA expression in the ganglion cells, inner nuclear layer cells, and pigment epithelium in the retina and in the nonpigment epithelium of the ciliary body. Immunoreactive ChM-I was present in these cells and also in the vitreous body. Western blot analysis detected an approximately 25-kDa band of ChM-I presumed as a secretory form in the aqueous humor and vitreous body and an approximately 37-kDa band as a precursor form in the retina. Recombinant human ChM-I inhibited tube morphogenesis of human retinal endothelial cells in vitro.
These observations indicate a potential role for ChM-I in inhibition of angiogenesis in the rat eye.
软骨调节素（ChM）-I在软骨中被鉴定为血管生成抑制剂，本研究旨在确定其在眼中的作用，研究ChM-I的表达、定位及可能的功能。
采用核糖核酸酶保护分析、原位杂交及免疫染色法，使用抗合成肽抗体检测大鼠眼中ChM-I的表达和定位。在培养中检测重组ChM-I对视网膜内皮细胞管形态发生的影响。
确定大鼠ChM-I基因编码334个氨基酸残基的开放阅读框，ChM-I mRNA仅在大鼠的软骨、眼和小脑中表达。ChM-I mRNA在虹膜睫状体、视网膜和巩膜部分有明显表达，但在眼的其他部分未表达。原位杂交显示视网膜中的神经节细胞、内核层细胞和色素上皮以及睫状体的非色素上皮中有mRNA表达。免疫反应性ChM-I存在于这些细胞以及玻璃体中。蛋白质印迹分析在房水和玻璃体中检测到一条约25 kDa的ChM-I条带，推测为分泌形式，在视网膜中检测到一条约37 kDa的条带，为前体形式。重组人ChM-I在体外抑制人视网膜内皮细胞的管形态发生。
这些观察结果表明ChM-I在抑制大鼠眼血管生成中具有潜在作用。
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