Fitch J M, Linsenmayer T F
Dev Biol. 1983 Jan;95(1):137-53. doi: 10.1016/0012-1606(83)90013-1.
As a first step in a study of the role(s) of basement membranes in ocular morphogenesis, we have produced a variety of monoclonal antibodies against native lens capsule from adult chicks, and have used these reagents to stain histological sections of ocular tissues from 4 1/2- to 18-day-old chicken embryos. Four different patterns of immunofluorescence were observed in sections of corneas of 18-day-old chicken embryos stained with these antibodies. The antibodies in group 1 stained the basement membranes of both the corneal epithelium and the endothelium (as well as Descemet's membrane). Those in groups 2 and 3 stained only the epithelial or endothelial basement membranes, respectively. The group 4 antibody stained the corneal stroma as well as Bowman's membrane and Descemet's membrane. The antibodies in group 1 could be further subdivided into groups 1a and 1b on the basis of temporal differences in the onset of staining in corneas from 4 1/2- to 7-day-old embryos. Thus, this series of monoclonal antibodies appears to recognize at least five different antigenic determinants. When these antibodies were used to stain sections of eyes at different stages of development, we found that the characteristic differential staining of some basement membranes was maintained throughout development, while the staining properties of others changed. This indicates that many of the ocular basement membranes may differ from one another in composition or conformation, and that at least some of them may undergo developmental changes. We also noticed a similarity in the pattern of fluorescence associated with the basement membranes of the limbal blood vessels and the corneal endothelium that is consistent with the hypothesis that the corneal endothelium is derived from the early periocular vascular endothelium. Our observations of developing corneas also revealed that the antigen recognized by the group 4 antibody may be produced by both the corneal epithelium and the stromal fibroblasts. The suitability of monoclonal antibodies for probing basement membrane heterogeneity is discussed.
作为研究基底膜在眼形态发生中作用的第一步，我们制备了多种针对成年鸡天然晶状体囊膜的单克隆抗体，并使用这些试剂对4.5至18日龄鸡胚的眼组织组织切片进行染色。在用这些抗体染色的18日龄鸡胚角膜切片中观察到四种不同的免疫荧光模式。第1组抗体对角膜上皮和内皮的基底膜（以及后弹力层）进行染色。第2组和第3组抗体分别仅对上皮或内皮基底膜进行染色。第4组抗体对角膜基质以及Bowman膜和后弹力层进行染色。根据4.5至7日龄胚胎角膜染色开始时间的差异，第1组抗体可进一步细分为1a组和1b组。因此，这一系列单克隆抗体似乎识别至少五种不同的抗原决定簇。当使用这些抗体对不同发育阶段的眼睛切片进行染色时，我们发现一些基底膜的特征性差异染色在整个发育过程中保持不变，而其他基底膜的染色特性则发生了变化。这表明许多眼基底膜在组成或构象上可能彼此不同，并且其中至少一些可能经历发育变化。我们还注意到与角膜缘血管和角膜内皮基底膜相关的荧光模式相似，这与角膜内皮源自早期眼周血管内皮的假设一致。我们对发育中角膜的观察还表明，第4组抗体识别的抗原可能由角膜上皮和基质成纤维细胞产生。讨论了单克隆抗体用于探测基底膜异质性的适用性。
