INVESTIGATION OF NEW MAMBRANE MUCIN PRODUCED BY THE OCULAR SURFACE EPITHELIUM

眼表上皮产生的新膜粘蛋白的研究

• 批准号: 13671838
• 负责人: WATANABE Hiroshi
• 金额: $ 2.24万
• 依托单位: Osaka University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-13671838/
• 关键词: mucin; cornea; conjunctiva; Mucin; 角膜上皮; IFN-g

MUC 1, 4 and 5AC have been reported to express in the cornea and the conjunctiva. There exists some trans - membrane mucins capable of maintaining tear film over the ocular surface epithelium. However, based on knock out studies in mice, there are other mucins than MUC 1, and MUC4 having the function. Recently, MUC16 has been reported and this MUC16 may have the function to maintain tear film over the ocular surface. By real-time PCR on reverse transcription products from RNA isolated from human corneal and conjunctival tissues, MUC16 mucin gene was determined. Further studies by in situ hybridization and immunohistochemistry disclosed MUC16 mRNA was localized to the apical cell layers of the cornea. MUC16 protein was obsertved in apical cells of the corneal stratified cells. Whether MUC1 and MUC16 mucins carry the H185 carbohydrate epitope was performed by one- or two-step immunoprecipitation assays and immunodepletion experiments followed by Western blot analysis. Immunofluorescence microscopy demonstrated exact colocalization of the MUC16 mucin and the H185 carbohydrate epitope in the cornea. Immunoprecipitated products recognized by the H185 antibody mucin was recognized by the antibody recognizing MUC16 and Immunoprecipitated MUC16 mucin was recognized by the H185 antibody. These studies showed that MUC16 mucin carries the H185 epitope. Immunodepletion with H185 antibody has no reactivity to the antibodies by MUC16. No cross-reactivity between immunoprecipitated MUC1 and the H185 antibody was disclosed. This study demonstrates that the membrane-associated mucin MUC16 is expressed by the corneal epithelium. Moreover, MUC16 carries the H185 carbohydrate epitope.

据报道，MUC 1、4和5AC在角膜和结膜中表达。在眼表上皮上存在一些能维持泪膜的跨膜粘蛋白。然而，根据对小鼠的敲除研究，除了muc1和MUC4之外，还有其他粘蛋白具有这种功能。最近，MUC16被报道，该MUC16可能具有维持眼表泪膜的功能。通过对人角膜和结膜组织RNA反转录产物的实时荧光定量PCR检测MUC16粘蛋白基因。通过原位杂交和免疫组织化学进一步研究发现MUC16 mRNA定位于角膜的顶端细胞层。在角膜分层细胞的顶端细胞中可见MUC16蛋白。MUC1和MUC16粘蛋白是否携带H185碳水化合物表位通过一步或两步免疫沉淀试验和免疫消耗实验进行，然后进行Western blot分析。免疫荧光显微镜显示MUC16粘蛋白和H185碳水化合物表位在角膜中精确共定位。被H185抗体识别的免疫沉淀产物被识别MUC16的抗体识别，免疫沉淀MUC16的粘蛋白被H185抗体识别。这些研究表明MUC16粘蛋白携带H185表位。MUC16对H185抗体无反应性。免疫沉淀MUC1与H185抗体之间无交叉反应性。本研究表明，膜相关黏液蛋白MUC16在角膜上皮中表达。此外，MUC16携带H185碳水化合物表位。

项目成果

蛍光濾過フィルターを用いた角結膜フルオレセイン染色所見の有効な観察法

使用荧光滤光片观察角结膜荧光素染色结果的有效方法

• 发表时间: 2003
• 期刊: 日本眼科紀要 54
• 作者: 坂東勝美;西田幸二;前田直之;渡辺仁;田野保雄
• 通讯作者: 田野保雄

Diagnosing dry eye using a blue-free barrier filter

• DOI: 10.1016/s0002-9394(03)00317-9
• 发表时间: 2003-09-01
• 期刊: AMERICAN JOURNAL OF OPHTHALMOLOGY
• 影响因子: 4.2
• 作者: Koh, S;Watanabe, H;Tano, Y
• 通讯作者: Tano, Y

Bessho K, Maeda N, Watanabe H et al.: "Fourier analysis of corneal astigmatic changes following photorefractive keratectomy"Seminar in Ophthalmology. 18(1). 23-28 (2003)

Bessho K、Maeda N、Watanabe H 等人：“屈光性角膜切除术后角膜散光变化的傅立叶分析”眼科研讨会。

Usefulness of a Fluorescence Barrier Filter For Fluorescein Staining of the Ocular Surface.

荧光屏障滤光片对于眼表面荧光素染色的有用性。

• 发表时间: 2003
• 期刊: Folia Ophthalmologica Japonica 54
• 作者: Bando K;Nishida K;Maeda N;Watanabe H;Tano Y.
• 通讯作者: Tano Y.

Koh S, Watanabe H, et al.: "Diagnosing dry eye using a blue-free barrier filter"American Journal of Ophthalmology. 136(3). 513-519 (2003)

Koh S、Watanabe H 等人：“使用无蓝光屏障滤光片诊断干眼症”美国眼科杂志。