Chidlow Glyn, Wood John P M, Knoops Bernard, Casson Robert J
Ophthalmic Research Laboratories, South Australian Institute of Ophthalmology, Hanson Institute Centre for Neurological Diseases, Frome Rd, Adelaide, SA, 5000, Australia.
Department of Ophthalmology and Visual Sciences, University of Adelaide, Frome Rd, Adelaide, SA, 5000, Australia.
Brain Struct Funct. 2016 Nov;221(8):3903-3925. doi: 10.1007/s00429-015-1135-3. Epub 2015 Oct 26.
Oxidative stress is implicated in various pathological conditions of the retina and optic nerve. Peroxiredoxins (Prdxs) comprise a recently characterized family of antioxidant enzymes. To date, little information exists regarding the distribution of Prdxs in the eye. Herein, we employed a combination of qRT-PCR, immunohistochemistry and Western blotting to determine the level of expression and distribution of the six Prdx isoforms in the retina and optic nerve of the rat. In addition, we performed some parallel analyses on the common marmoset (Callithrix Jacchus). In the rat, all of the Prdx transcripts were expressed in relatively high amounts in both retina and optic nerve, with abundances ranging from approximately 3-50 % of the level of the housekeeping gene cyclophilin. With regard to protein expression, each isoform was detected in the retina and optic nerve by either Western blotting and/or immunohistochemistry. Excepting Prdx4, there was a good correspondence between the rodent and primate results. In the retina, Prdx1 and Prdx2 were principally localized to neurons in the inner nuclear layer and cone photoreceptors, Prdx3 and Prdx5 displayed characteristic mitochondrial immunolabeling, while Prdx6 was associated with astrocytes and Müller cells. In the optic nerve, Prdx1 was robustly expressed by oligodendrocytes, Prdx3 and Prdx5 were observed in axons, and Prdx6 was restricted to astrocytes. The present findings augment our understanding of the distribution and expression of the Prdxs in the retina and optic nerve of rodents and primates and lay the foundation for subsequent analysis of their involvement in relevant blinding diseases.
氧化应激与视网膜和视神经的多种病理状况有关。过氧化物酶(Prdxs)是最近被鉴定出的一类抗氧化酶家族。迄今为止,关于Prdxs在眼部的分布情况所知甚少。在此,我们采用定量逆转录聚合酶链反应(qRT-PCR)、免疫组织化学和蛋白质免疫印迹法相结合的方法,来确定大鼠视网膜和视神经中六种Prdx亚型的表达水平和分布情况。此外,我们还对普通狨猴(Callithrix Jacchus)进行了一些平行分析。在大鼠中,所有Prdx转录本在视网膜和视神经中均有较高表达,丰度范围约为管家基因亲环蛋白水平的3%-50%。在蛋白质表达方面,通过蛋白质免疫印迹法和/或免疫组织化学在视网膜和视神经中检测到了每种亚型。除Prdx4外,啮齿动物和灵长类动物的结果有较好的一致性。在视网膜中,Prdx1和Prdx2主要定位于内核层的神经元和视锥光感受器,Prdx3和Prdx5呈现出线粒体特征性免疫标记,而Prdx6与星形胶质细胞和穆勒细胞有关。在视神经中,少突胶质细胞强烈表达Prdx1,在轴突中观察到Prdx3和Prdx5,而Prdx6仅限于星形胶质细胞。本研究结果加深了我们对Prdxs在啮齿动物和灵长类动物视网膜和视神经中的分布及表达的理解,并为后续分析它们在相关致盲疾病中的作用奠定了基础。