Kwong Jacky M K, Caprioli Joseph
Department of Ophthalmology, Jules Stein Eye Institute, David Geffen School of Medicine at University of California Los Angeles, Room B-146, 100 Stein Plaza, Los Angeles, CA 90095-7000, USA.
Exp Eye Res. 2006 Apr;82(4):576-82. doi: 10.1016/j.exer.2005.08.017. Epub 2005 Sep 28.
To examine the expression of phosphorylated c-Jun N-terminal kinase (JNK) in cells in the retinal ganglion cell layer of glaucoma, intraocular pressure (IOP) of adult Wistar rats was elevated unilaterally by repeated trabecular argon laser photocoagulation 5 days after intracameral injection of India ink. Animals were euthanized after 3 days, and 1, 2, and 5 weeks of IOP elevation. Immunohistochemistry with specific antibodies against phosphorylated JNK was performed on retinas. Retrograde labeling using Fluorogold and fluorescence immunohistochemistry was performed on retinas 5 weeks after IOP elevation. TdT-mediated biotin-dUTP nick end labeling (TUNEL) was performed on the retinal sections to determine the rate of cell death. There was increased IOP (52.3%) from 3 days to 5 weeks after repeated trabecular laser photocoagulation. Mean number of TUNEL-positive cells in the retinal ganglion cell layer of eyes with experimental glaucoma was 0.43, 0.36, 0.57, and 0.19 per retinal section at 3 days, and 1, 2 and 5 weeks, respectively. No TUNEL-positive cells were noted in controls. In parallel to TUNEL, significantly increased numbers of phosphorylated JNK-labeled cells in the retinal ganglion cell layer were noted at 3 days (9.95 versus 4.15; P=0.005), 1 week (7.65 versus 4.00; P=0.006), 2 weeks (9.13 versus 4.48; P=0.032), and 5 weeks (8.06 versus 4.96; P=0.017) of IOP elevation when compared with contralateral control eyes. Fluorogold labeled RGCs were co-localized with increased phosphorylated JNK immunoreactivity. Some TUNEL-positive cells were phosphorylated JNK immuno-positive. Phosphorylation of JNK occurs in experimental glaucoma and may play a role in retinal ganglion cell death.
为检测青光眼视网膜神经节细胞层中细胞内磷酸化c-Jun氨基末端激酶（JNK）的表达，在成年Wistar大鼠前房注射印度墨汁5天后，通过重复小梁氩激光光凝术单侧升高眼压。在眼压升高3天、1周、2周和5周后对动物实施安乐死。使用抗磷酸化JNK的特异性抗体对视网膜进行免疫组织化学检测。在眼压升高5周后，使用荧光金进行逆行标记并对视网膜进行荧光免疫组织化学检测。对视网膜切片进行TdT介导的生物素-dUTP缺口末端标记（TUNEL）以确定细胞死亡率。重复小梁激光光凝术后3天至5周眼压升高（52.3%）。实验性青光眼患眼视网膜神经节细胞层中TUNEL阳性细胞的平均数量在3天、1周、2周和5周时分别为每视网膜切片0.43、0.36、0.57和0.19个。对照组未发现TUNEL阳性细胞。与TUNEL检测结果一致，与对侧对照眼相比，在眼压升高3天（9.95对4.15；P=0.005）、1周（7.65对4.00；P=0.006）、2周（9.13对4.48；P=0.032）和5周（8.06对4.96；P=0.017）时，视网膜神经节细胞层中磷酸化JNK标记的细胞数量显著增加。荧光金标记的视网膜神经节细胞与增强的磷酸化JNK免疫反应性共定位。一些TUNEL阳性细胞为磷酸化JNK免疫阳性。JNK的磷酸化发生在实验性青光眼中，可能在视网膜神经节细胞死亡中起作用。
