Mohamed Yasser Helmy, Uematsu Masafumi, Kusano Mao, Inoue Daisuke, Tang Diya, Suzuki Keiji, Kitaoka Takashi
Department of Ophthalmology and Visual Sciences, Graduate School of Biomedical Sciences, Nagasaki University, Nagasaki 852-8501, Japan.
Department of Radiation Medical Sciences, Atomic Bomb Disease Institute, Nagasaki University, Nagasaki 852-8523, Japan.
Life (Basel). 2024 Aug 22;14(8):1046. doi: 10.3390/life14081046.
We developed a technique that can measure corneal transepithelial electrical resistance (TER) in mice, which was used for evaluating corneal toxicity induced by ophthalmic drugs. We used a tissue culture well and its insert to mount the mouse globe and separated the cornea from the rest of the globe to enable corneal TER measurements to be taken. The explanted mouse eyes were divided into groups, and the corneal epithelia were exposed to different concentrations of BAC. Half of these eyes were fixed for transmission electron microscopy (TEM) examination and the other for ZO-1 immunohistochemical (IHC) evaluation. After exposure to control, 0.1%, 0.2%, and 0.5% BAC, the TER was 100 ± 0%, 91 ± 14%, 83 ± 13%, and 34 ± 12% of the pre-exposure TER at 1 min, respectively, with a statistically significant decrease in the 0.5% group. After 3 min, the TER showed a statistically significant decrease in the 0.2% and 0.5% groups. The TEM examinations showed a loss of epithelial tight junctions between superficial cells in the 0.2% and 0.5% groups. The IHC examination showed decreased ZO-1 staining of the corneal epithelium of the same groups as compared to the control. To the best of our knowledge, we succeeded in developing an innovative technique for corneal TER measurement in mice.
我们开发了一种可测量小鼠角膜跨上皮电阻（TER）的技术，该技术用于评估眼科药物引起的角膜毒性。我们使用组织培养孔及其插入物来固定小鼠眼球，并将角膜与眼球的其余部分分离，以便能够进行角膜TER测量。将取出的小鼠眼睛分组，使角膜上皮暴露于不同浓度的苯扎氯铵（BAC）。这些眼睛的一半用于透射电子显微镜（TEM）检查，另一半用于ZO-1免疫组织化学（IHC）评估。在暴露于对照、0.1%、0.2%和0.5%的BAC后，在1分钟时，TER分别为暴露前TER的100±0%、91±14%、83±13%和34±12%，0.5%组有统计学意义的下降。3分钟后，0.2%和0.5%组的TER有统计学意义的下降。TEM检查显示0.2%和0.5%组的表层细胞之间上皮紧密连接丧失。IHC检查显示，与对照组相比，相同组的角膜上皮ZO-1染色减少。据我们所知，我们成功开发了一种用于测量小鼠角膜TER 的创新技术。
