Wang An-Guor, Lee Chia-Ming, Wang Yu-Chuan, Lin Chi-Hung, Fann Ming-Ji
Department of Ophthalmology, Taipei Veterans General Hospital, Taipei, Taiwan, Republic of China.
Exp Eye Res. 2002 May;74(5):651-9. doi: 10.1006/exer.2002.1190.
The purpose of this study is to investigate the cytochrome oxidase (COX) activity in the retina and optic nerve following an optic nerve injury. The optic nerve crush of one eye was carried out in Balb/c mice. A semi-quantitative RT-PCR method was then adopted to evaluate the mRNA expression of cytochrome oxidase subunit 1 (COX1) in the retina after surgery. Up-regulation of COX1 mRNA in the retina was detected by RT-PCR at 24 hr following the optic nerve injury. Total retinal mitochondrial mass measured by fluorescent intensity of MitoTracker green was not altered following the injury. COX histochemistry performed on cryostat sections showed an elevated enzyme activity of COX in the retina and in the optic nerve. In the retina, elevation of the COX activity was observed in the retinal ganglion cell layer and the overlying nerve fibre layer. The increase of COX activity began from 24 hr after injury, peaked around day 3, and maintained up to 1 week after the operation. In the optic nerve, increase of COX activity was observed in regions distal to the crush line and distributed either randomly or in a cone shape. In conclusion, both the expression of COX1 mRNA in retina and the activity of COX in inner plexiform layer and retinal ganglion cell layer were elevated following optic nerve injury without affecting total retinal mitochondrial mass. These findings suggested that one of early responses in the retina and in the optic nerve after the optic nerve injury is to scale up the energy production.
本研究的目的是调查视神经损伤后视网膜和视神经中的细胞色素氧化酶(COX)活性。在Balb/c小鼠中对一只眼睛进行视神经挤压。然后采用半定量RT-PCR方法评估手术后视网膜中细胞色素氧化酶亚基1(COX1)的mRNA表达。在视神经损伤后24小时通过RT-PCR检测到视网膜中COX1 mRNA上调。通过MitoTracker绿色荧光强度测量的视网膜线粒体总质量在损伤后未发生改变。在冷冻切片上进行的COX组织化学显示视网膜和视神经中COX的酶活性升高。在视网膜中,在视网膜神经节细胞层和上方的神经纤维层中观察到COX活性升高。COX活性的增加从损伤后24小时开始,在第3天左右达到峰值,并在手术后持续1周。在视神经中,在挤压线远端区域观察到COX活性增加,呈随机或锥形分布。总之,视神经损伤后视网膜中COX1 mRNA的表达以及内网状层和视网膜神经节细胞层中COX的活性均升高,而不影响视网膜线粒体总质量。这些发现表明,视神经损伤后视网膜和视神经的早期反应之一是扩大能量产生。