Zhang S R, Li S H, Abler A, Fu J, Tso M O, Lam T T
Georgiana Dvorak Theobald Ophthalmic Pathology Laboratory, Department of Ophthalmology and Visual Sciences, College of Medicine, University of Illinois at Chicago, USA.
Invest Ophthalmol Vis Sci. 1996 Aug;37(9):1793-9.
The possible involvement of tissue transglutaminase (tTG) in apoptosis during photoreceptor degeneration was examined in retinal photic injury in rats and in retinal dystrophy of Royal College of Surgeons (RCS) rats.
Retinal photic injury was induced in 48 male Lewis albino rats by exposure to green fluorescent light of 300 to 320 foot-candles. The retinal tTG was examined by enzyme assay, immunohistochemistry, and Western blot analysis after 9, 12, or 24 hours of exposure or at 6 or 24 hours of dark adaptation after 24 hours of light exposure. Retinas from RCS rats at various stages of degeneration also were examined with similar methods.
There was a progressive increase in retinal tTG activity after 300 to 320 ft-c of light exposure, reaching a peak after 24 hours of light exposure. In the RCS rats, tTG activity increased with age. Western blot analysis revealed an immunoreactive band at 80 kDa, which increased in accordance with the transglutaminase activity in both models. In normal rat retinas, tTG immunolabeling was present only in the outer segments. There was an increased number of immunolabeled photoreceptor nuclei from 12 hours of light exposure to 24 hours of light exposure. In the RCS rat, increasing numbers of immunopositive photoreceptor nuclei from 20 to 50 days of age were noted.
The data associated increased retinal tTG activity and enzyme levels with photoreceptor cells undergoing apoptosis. The tTG-dependent irreversible cross-linking of intracellular protein may play an important role in causing the structural changes in cells undergoing apoptosis in the retina.
在大鼠视网膜光损伤以及皇家外科学院(RCS)大鼠视网膜营养不良模型中,研究组织转谷氨酰胺酶(tTG)在光感受器变性过程中与细胞凋亡的可能关系。
对48只雄性Lewis白化大鼠进行视网膜光损伤,使其暴露于300至320英尺烛光的绿色荧光灯下。在光照9、12或24小时后,或在光照24小时后的暗适应6或24小时后,通过酶活性测定、免疫组织化学和蛋白质印迹分析检测视网膜tTG。对处于不同退变阶段的RCS大鼠视网膜也采用类似方法进行检测。
在300至320英尺烛光光照后,视网膜tTG活性逐渐增加,在光照24小时后达到峰值。在RCS大鼠中,tTG活性随年龄增长而增加。蛋白质印迹分析显示在80 kDa处有一条免疫反应条带,在两种模型中该条带均随转谷氨酰胺酶活性增加而增加。在正常大鼠视网膜中,tTG免疫标记仅存在于外段。从光照12小时到光照24小时,免疫标记的光感受器细胞核数量增加。在RCS大鼠中,从20至50日龄可见免疫阳性光感受器细胞核数量增加。
数据表明视网膜tTG活性和酶水平升高与经历凋亡的光感受器细胞有关。tTG依赖的细胞内蛋白质不可逆交联可能在导致视网膜中经历凋亡的细胞结构变化方面起重要作用。