Savontaus M, Ihanamäki T, Metsäranta M, Vuorio E, Sandberg-Lall M
Department of Medical Biochemistry, University of Turku, Finland.
Invest Ophthalmol Vis Sci. 1997 Apr;38(5):930-42.
To elucidate the function of type II collagen in the development and diseases of the eye by analyzing the temporospatial expression of the long (IIA) and short (IIB) isoforms of type II collagen in the normal and transgenic Dell mice.
Normal and Dell transgenic embryos harboring a deletion mutation in the pro alpha 1 (II) collagen chain were studied from day 10.5 of embryonic development up to day 10 postpartum. Northern and in situ hybridizations and RNase protection assays were used to study the developmental and temporospatial expression of type II collagen isoforms.
Expression of type II collagen mRNAs was observed at all developmental stages with maximum expression at 16.5 days of embryonic development. RNase protection analyses confirmed that both wild type and transgene-derived mRNAs underwent similar alternative splicing of exon 2 in the eye. By in situ hybridization, both isoforms were observed in the cornea, sclera, vitreous, ganglion cell layer of retina, developing ciliary body-iris, and in the retinal pigment epithelium-Bruch's membrane as well as in the lens and conjunctiva. Differences were observed between eyes of Dell mice and of control subjects in the levels and temporal expression patterns of type II collagen mRNA, which resulted in structural abnormalities in histologic analysis.
Widespread expression of type II collagen mRNAs in ocular structures suggests an important role for type II collagen in structural development of the eye. As the expression patterns observed correspond to structural abnormalities in the eyes of Dell mice, the current results offer a promising basis for further development of mouse models for arthroophthalmopathies.
通过分析正常和转基因戴尔小鼠中Ⅱ型胶原蛋白长（IIA）和短（IIB）异构体的时空表达，阐明Ⅱ型胶原蛋白在眼睛发育和疾病中的作用。
研究携带α1（II）前胶原链缺失突变的正常和戴尔转基因胚胎，从胚胎发育第10.5天到产后第10天。采用Northern杂交、原位杂交和核糖核酸酶保护分析来研究Ⅱ型胶原蛋白异构体的发育和时空表达。
在所有发育阶段均观察到Ⅱ型胶原蛋白mRNA的表达，在胚胎发育第16.5天时表达量最高。核糖核酸酶保护分析证实，野生型和转基因来源的mRNA在眼中经历了相似的外显子2可变剪接。通过原位杂交，在角膜、巩膜、玻璃体、视网膜神经节细胞层、发育中的睫状体 - 虹膜、视网膜色素上皮 - 布鲁赫膜以及晶状体和结膜中均观察到两种异构体。在戴尔小鼠和对照受试者的眼中，Ⅱ型胶原蛋白mRNA的水平和时间表达模式存在差异，这导致了组织学分析中的结构异常。
Ⅱ型胶原蛋白mRNA在眼部结构中的广泛表达表明Ⅱ型胶原蛋白在眼睛结构发育中起重要作用。由于观察到的表达模式与戴尔小鼠眼睛中的结构异常相对应，目前的结果为进一步开发关节眼病小鼠模型提供了有前景的基础。
