Rutar Matt, Natoli Riccardo, Chia R X, Valter Krisztina, Provis Jan M
John Curtin School of Medical Research, The Australian National University, Building 131, Garran Road, Canberra, ACT 2601, Australia.
ANU Medical School, The Australian National University, 54 Mills Road, Canberra, ACT 2601, Australia.
J Neuroinflammation. 2015 Jan 17;12:8. doi: 10.1186/s12974-014-0224-1.
Monocyte infiltration is involved in the pathogenesis of many retinal degenerative conditions. This process traditionally depends on local expression of chemokines, though the roles of many of these in the degenerating retina are unclear. Here, we investigate expression and in situ localization of the broad chemokine response in a light-induced model of retinal degeneration.
Sprague-Dawley (SD) rats were exposed to 1,000 lux light damage (LD) for up to 24 hrs. At time points during (1 to 24 hrs) and following (3 and 7 days) exposure, animals were euthanized and retinas processed. Microarray analysis assessed differential expression of chemokines. Some genes were further investigated using polymerase chain reaction (PCR) and in situ hybridization and contrasted with photoreceptor apoptosis using terminal deoxynucleotidyl transferase dUTP nick-end labeling (TUNEL). Recruitment of retinal CD45 (+) leukocytes was determined via fluorescence activated cell sorting (FACS), and expression of chemokine receptors determined using PCR.
Exposure to 24 hrs of LD resulted in differential expression of chemokines including Ccl3, Ccl4, Ccl7, Cxcl1, and Cxcl10. Their upregulation correlated strongly with peak photoreceptor death, at 24 hrs exposure. In situ hybridization revealed that the modulated chemokines were expressed by a combination of Müller cells, activated microglia, and retinal pigment epithelium (RPE). This preceded large increases in the number of CD45(+) cells at 3- and 7-days post exposure, which expressed a corresponding repertoire of chemokine receptors.
Our data indicate that retinal degeneration induces upregulation of a broad chemokine response whose expression is coordinated by Müller cells, microglia, and RPE. The findings inform our understanding of the processes govern the trafficking of leukocytes, which are contributors in the pathology of retinal degenerations.
单核细胞浸润参与多种视网膜退行性疾病的发病机制。传统上，这一过程依赖趋化因子的局部表达，尽管其中许多趋化因子在退行性视网膜中的作用尚不清楚。在此，我们在光诱导的视网膜变性模型中研究广泛趋化因子反应的表达及原位定位。
将Sprague-Dawley（SD）大鼠暴露于1000勒克斯的光损伤（LD）下长达24小时。在暴露期间（1至24小时）及暴露后（3天和7天）的时间点，对动物实施安乐死并处理视网膜。通过微阵列分析评估趋化因子的差异表达。使用聚合酶链反应（PCR）和原位杂交进一步研究一些基因，并与使用末端脱氧核苷酸转移酶dUTP缺口末端标记（TUNEL）的光感受器凋亡进行对比。通过荧光激活细胞分选（FACS）确定视网膜CD45（+）白细胞的募集情况，并使用PCR确定趋化因子受体的表达。
暴露于24小时的光损伤导致趋化因子Ccl3、Ccl4、Ccl7、Cxcl1和Cxcl10的差异表达。它们的上调与暴露24小时时光感受器死亡高峰密切相关。原位杂交显示，受调节的趋化因子由穆勒细胞、活化的小胶质细胞和视网膜色素上皮（RPE）共同表达。这在暴露后3天和7天CD45（+）细胞数量大幅增加之前出现，这些细胞表达相应的趋化因子受体库。
我们的数据表明，视网膜变性诱导广泛趋化因子反应的上调，其表达由穆勒细胞、小胶质细胞和RPE协调。这些发现有助于我们理解调控白细胞 trafficking的过程，白细胞是视网膜退行性病变病理学的促成因素。
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