Szerenyi K, Wang X, Gabrielian K, LaBree L, McDonnell P J
Doheny Eye Institute, Los Angeles, CA 90033.
Cornea. 1994 Nov;13(6):487-92.
The purpose of this study was to determine whether the DNA of mitotic cells in the corneal and conjunctival epithelium can be labeled with 5-bromo-2-deoxyuridine (BrdU) immunostaining. Both corneas of four New Zealand white rabbits were deepithelialized in the center and the regenerated epithelium was evaluated for mitosis at 1, 3, 6, and 10 days. Unwounded corneas of three rabbits were labeled for baseline measurements. We administered the marker intravenously to all seven rabbits 15 h before scheduled killing. Immediately after killing, all of the globes were enucleated and histologic sections were prepared. In unwounded corneas, labeled cells were quantitated and the fraction of mitotic cells in the center of the cornea, in the periphery of the cornea, and in the conjunctiva were compared. In deepithelialized corneas, increase in mitosis in the central epithelium was quantitated. All of the unwounded eyes showed mitosis in the basal layer of both the corneal and conjunctival epithelium. In the center of the cornea 4.1 +/- 2.9% of the epithelial cells were labeled, in the corneal periphery 4.3 +/- 1.7% of the cells were labeled, and in the conjunctiva 4.1 +/- 1.9% of the epithelial cells were labeled, with a p value ranging from 0.84 to 0.99. In wounded corneas, when compared with cell counts in unwounded eyes, 52.6% of the epithelial cells were in mitosis on day 3, 13.9% were in mitosis on day 6, and by day 10 baseline values of 4.2% were obtained. We conclude that BrdU immunostaining is a safe, efficient, and less costly alternative to autoradiography for visualization of dividing corneal and conjunctival epithelium.
本研究的目的是确定角膜和结膜上皮有丝分裂细胞的DNA是否能用5-溴-2-脱氧尿苷（BrdU）免疫染色进行标记。对4只新西兰白兔的双眼角膜中央进行上皮剥除，并在第1、3、6和10天评估再生上皮的有丝分裂情况。对3只兔子未受伤的角膜进行标记以作基线测量。在预定处死前15小时，对所有7只兔子静脉注射该标记物。处死兔子后，立即摘除所有眼球并制备组织学切片。在未受伤的角膜中，对标记细胞进行定量，并比较角膜中央、角膜周边和结膜中有丝分裂细胞的比例。在角膜上皮剥除的角膜中，对中央上皮有丝分裂的增加情况进行定量。所有未受伤的眼睛在角膜和结膜上皮的基底层均显示有丝分裂。在角膜中央，4.1±2.9%的上皮细胞被标记，在角膜周边，4.3±1.7%的细胞被标记，在结膜中，4.1±1.9%的上皮细胞被标记，p值范围为0.84至0.99。在受伤的角膜中，与未受伤眼睛的细胞计数相比，第3天52.6%的上皮细胞处于有丝分裂状态，第6天13.9%处于有丝分裂状态，到第10天获得了4.2%的基线值。我们得出结论，对于可视化角膜和结膜上皮的分裂情况，BrdU免疫染色是一种安全、高效且成本较低的放射自显影替代方法。
