Osborne N N, Larsen A K
Nuffield Laboratory of Ophthalmology, University of Oxford, U.K.
Neurochem Int. 1996 Sep;29(3):263-70. doi: 10.1016/0197-0186(96)00005-8.
Raising the rat's intraocular pressure above the systolic blood pressure for 60 min followed by a reperfusion of 7-10 days caused an ischaemic insult to the retina. The b-wave of the electroretinogram was abolished and the retinal thickness was much reduced, the greatest influence being associated with the inner retinal layers. No obvious histological damage was apparent at the light microscopy level. Immunocytochemistry, however, revealed a clear change in the nature of certain antigens associated with specific cell-types. Thy-1 antigen located to ganglion cell membranes was much reduced, suggesting that the ganglion cells are affected by ischaemia. Calretinin-immunoreactivity associated with amacrine cells is drastically reduced by ischaemia. In contrast, Ret-P1, located to the outer segments of the photoreceptors is unaffected by ischaemia. Ischaemia also caused GFAP-immunoreactivity to be expressed in the Müller cells, which is normally only associated with astrocytes in the ganglion/nerve fibre layer. Injection of a mixture of CNQX and MK-801, kainate and NMDA receptor antagonists, respectively, into the eye just before ischaemia failed to reverse the changes induced by the insult. However, analysis 3 days after reperfusion revealed that when the ischaemic insult was reduced to 45, rather than 60 min, the changes in the calretinin-immunoreactivity were reversed. The results show that immunocytochemistry provides a powerful way of following biochemical changes associated with specific cell types caused by ischaemia.
将大鼠眼压升高至收缩压以上并持续60分钟,随后进行7 - 10天的再灌注,会对视网膜造成缺血性损伤。视网膜电图的b波消失,视网膜厚度大幅减小,其中对内视网膜层的影响最大。在光学显微镜水平未观察到明显的组织学损伤。然而,免疫细胞化学显示与特定细胞类型相关的某些抗原的性质发生了明显变化。位于神经节细胞膜上的Thy - 1抗原显著减少,表明神经节细胞受到缺血影响。与无长突细胞相关的钙视网膜蛋白免疫反应性因缺血而急剧降低。相比之下,位于光感受器外段的Ret - P1不受缺血影响。缺血还导致穆勒细胞中出现GFAP免疫反应性,而GFAP通常仅与神经节/神经纤维层中的星形胶质细胞相关。在缺血前向眼内注射分别为AMPA和NMDA受体拮抗剂的CNQX和MK - 801混合物,未能逆转损伤诱导的变化。然而,再灌注3天后的分析显示,当缺血损伤时间缩短至45分钟而非60分钟时,钙视网膜蛋白免疫反应性的变化得以逆转。结果表明,免疫细胞化学为追踪缺血引起的与特定细胞类型相关的生化变化提供了一种有力的方法。