Laboratory of the Biology and Pathology of the Eye, Institute of Inherited Metabolic Disorders, General Teaching Hospital and Charles University, Prague, Czech Republic.
Histol Histopathol. 2011 Mar;26(3):323-31. doi: 10.14670/HH-26.323.
The aim of this study was to detect a spectrum of cytokeratins (CK) present in the adult human cornea, limbus and perilimbal conjunctiva. Cryosections from seven corneo-scleral discs were fixed, and indirect immunofluorescent staining was performed using antibodies directed against CK1-CK10 and CK13-CK20. The percentage of positive cells was calculated in the epithelium of the cornea, limbus and perilimbal conjunctiva. Quantitative real time RT-PCR (qRT-PCR) was used to detect CK6 and CK18 expression in the corneal and conjunctival epithelium. The most intense staining present throughout the cornea was observed for CK3, CK5 and CK14; CK19 was found at the corneal periphery only. CK4 and CK10/13 revealed mild to moderate positivity mostly in the superficial layers of the cornea. The suprabasal cell layers of all examined areas showed a strong positivity for CK16. A heterogeneous staining pattern with a centrifugal decrease in the signal was observed for CK8 and CK18. CK5/6, CK14 and CK19 were present in the limbus, where a positive signal for CK3 was observed in the suprabasal and superficial cells only. In contrast to the cornea, CK15 appeared in the basal and suprabasal layers of the limbus. The perilimbal conjunctiva showed strong immunostaining for CK10/13, CK14 and CK19. A moderate signal for CK7 was detected in the superficial layers of the conjunctiva. qRT-PCR confirmed CK6 and CK18 expression in the corneal and conjunctival epithelium. The detailed characterization of the corneal, limbal and perilimbal conjunctival epithelium under normal circumstances may be useful for characterizing the changes occurring under pathological conditions.
本研究旨在检测成人眼角膜、角膜缘和周边部球结膜中存在的细胞角蛋白（CK）谱。将七个角巩膜环切片固定，使用针对 CK1-CK10 和 CK13-CK20 的抗体进行间接免疫荧光染色。计算角膜、角膜缘和周边部球结膜上皮中阳性细胞的百分比。使用定量实时 RT-PCR（qRT-PCR）检测角膜和结膜上皮中 CK6 和 CK18 的表达。在整个角膜中观察到 CK3、CK5 和 CK14 存在最强的染色；仅在角膜周边观察到 CK19。CK4 和 CK10/13 主要在角膜浅层显示出轻度至中度阳性。所有检查区域的基底上层细胞均强烈表达 CK16。观察到 CK8 和 CK18 的异质染色模式，信号呈离心性降低。CK5/6、CK14 和 CK19 存在于角膜缘，仅在上皮的基底上层观察到 CK3 的阳性信号。与角膜相反，CK15 出现在角膜缘的基底和基底上层。周边球结膜强烈表达 CK10/13、CK14 和 CK19。在结膜的浅层检测到 CK7 的中度信号。qRT-PCR 证实了角膜和结膜上皮中 CK6 和 CK18 的表达。正常情况下，对角膜、角膜缘和周边球结膜上皮的详细特征描述可能有助于描述病理条件下发生的变化。
