Perlecan-Deficient Mutation Impairs Corneal Epithelial Structure
Perlecan-Deficient Mutation Impairs Corneal Epithelial Structure
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基底膜蛋白缺陷突变损害角膜上皮结构
DOI：
10.1167/iovs.11-8742
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发表时间：
2012-03-01
影响因子：
4.4
通讯作者：
Arikawa-Hirasawa, Eri
中科院分区：
文献类型：
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作者：
Inomata, Takenori;Ebihara, Nobuyuki;Arikawa-Hirasawa, Eri
目的。阐明基底膜蛋白聚糖 (Hspg2)（一种在基底膜中表达的大型多结构域硫酸乙酰肝素蛋白多糖）在角膜上皮结构中的作用。方法。使用先前开发的基底膜蛋白缺陷（Hspg2(-/-)-Tg）小鼠模型。通过光学和透射电子显微镜对他们的角膜进行组织学分析。通过免疫组织化学检查基底膜蛋白聚糖在野生型（WT）小鼠和Hspg2（-/-）-Tg 小鼠角膜中的定位。通过免疫组化和实时聚合酶链反应（PCR）分析基底膜蛋白缺陷对角膜上皮结构的影响，包括Ki67、细胞角蛋白12（K12）、连接蛋白43（Cx43）、Notch1和Pax6等角膜上皮增殖和分化标志物的表达。与WT小鼠相比，Hspg2(-/-)-Tg小鼠有小眼球和更薄的角膜上皮。基底膜蛋白聚糖位于 WT 小鼠的角膜上皮基底膜中，但不在 Hspg2(-/-)-Tg 小鼠中。与WT角膜上皮相比，Hspg2(-/-)-Tg角膜上皮表现出更薄的翼细胞层和减少的K167阳性细胞数量，但没有死亡细胞。免疫组织化学和实时PCR分析显示，与WT小鼠相比，Hspg2(-/-)-Tg小鼠中角膜上皮分化标志物如K12、Cx43、Notch1和Pax6的表达显着降低。结论。这项研究的结果强调了基底膜中基底膜蛋白聚糖的存在与角膜上皮结构之间的密切相关性，并且基底膜蛋白聚糖缺陷突变会损害角膜上皮结构。 （投资眼科可见科学。2012 年；53：1277-1284）DOI：10.1167/iovs.11-8742
PURPOSE. To elucidate the role of perlecan (Hspg2), a large multidomain heparan sulfate proteoglycan expressed in the basement membrane, in the structure of the corneal epithelium.METHODS. A previously developed perlecan-deficient (Hspg2(-/-)-Tg) mouse model was used. Histologic analysis of their corneas was performed by light and transmission electron microscopy. The localization of perlecan in the corneas of wild-type (WT) mice and Hspg2(-/-)-Tg mice was examined by immunohistochemistry. The effects of perlecan deficiency on corneal epithelial structure was analyzed with respect to the expression of corneal epithelial proliferation and differentiation markers, such as Ki67, cytokeratin12 (K12), connexin43 (Cx43), Notch1, and Pax6 by immunohistochemistry and real-time polymerase chain reaction (PCR).RESULTS. The Hspg2(-/-)-Tg mice had microphthalmos and a thinner corneal epithelium compared with that of the WT mice. Perlecan was localized in the corneal epithelial basement membrane in the WT mice, but not in the Hspg2(-/-)-Tg mice. The Hspg2(-/-)-Tg corneal epithelium exhibited thinner wing cell layers and a decreased number of K167-positive cells, but no dead cells, compared with the WT corneal epithelium. Immunohistochemistry and real-time PCR analysis revealed a significantly decreased expression of corneal epithelial differentiation markers such as K12, Cx43, Notch1, and Pax6 in Hspg2(-/-)-Tg mice, compared with those of the WT mice.CONCLUSIONS. The findings of this study highlight a strong correlation between the presence of perlecan in the basement membrane and the structure of corneal epithelium and that the perlecan-deficient mutation impairs corneal epithelial structure. (Invest Ophthalmol Vis Sci. 2012;53:1277-1284) DOI:10.1167/iovs.11-8742