Human amniotic membrane, like corneal epithelial basement membrane, manifests the α5 chain of type IV collagen
Human amniotic membrane, like corneal epithelial basement membrane, manifests the α5 chain of type IV collagen
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DOI：
10.1167/iovs.03-0952
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发表时间：
2004-06-01
影响因子：
4.4
通讯作者：
Kinoshita, S
中科院分区：
文献类型：
--
作者：
Endo, K;Nakamura, T;Kinoshita, S
目的.为了重新检查W型胶原的α 5链（α 5 [IV]）是否被认为是缺失的，实际上存在于人羊膜中。对在剖腹产时获得的人羊膜的冷冻切片进行化学分析，以确定是否存在α 5（IV），包括或不包括变性步骤。用胶原酶消化羊膜以从a链释放非胶原NC1结构域。然后在Western印迹分析上测定α 5（IV）的NC1结构域。用从美国眼库获得的人角膜和结膜进行相同的实验。羊膜和角膜的变性样品的基底膜对α 5（W）染色呈阳性。在没有变性步骤的情况下，仅角膜样品为阳性。无论是否变性，结膜上皮均不染色。Western blot分析检测羊膜和角膜样本中α 5（IV）的NC 1结构域。羊膜的基底膜与角膜上皮的基底膜相似，而与结膜的基底膜不同。羊膜可能是角膜上皮细胞良好的基质。
PURPOSE. To reexamine whether the alpha5 chain of type W (alpha5[IV]) Collagen, thought to be absent, is in fact present in human amniotic membrane.METHODS. Cryosections of human amniotic membrane obtained at Cesarean section were immunohistochemically examined for the presence of alpha5(IV), with or without inclusion of the denaturing step. Amniotic membrane was digested with collagenase to release the noncollagenous NC1 domain from the a-chain. The NC1 domain of alpha5(IV) was then assayed on Western blot analysis. Identical experiments were performed with human corneas and conjunctivae obtained from an American eye bank.RESULTS. The basement membrane of denatured samples of amniotic membrane and cornea stained positive for alpha5(W). Without the denaturing step, only corneal samples were positive. With or without denaturing, conjunctival epithelium did not stain. Western blot analysis detected NC1 domains of alpha5(IV) in amniotic membrane and corneal samples.CONCLUSIONS. The basement membrane of amniotic membrane resembles that of corneal epithelium but not conjunctiva. Amniotic membrane may be an excellent substrate for corneal epithelial cells.