The Effects of Age and Cx3cr1 Deficiency on Retinal Microglia in the Ins2Akita Diabetic Mouse
The Effects of Age and Cx3cr1 Deficiency on Retinal Microglia in the Ins2Akita Diabetic Mouse
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DOI：
10.1167/iovs.12-10876
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发表时间：
2013-01-01
影响因子：
4.4
通讯作者：
McMenamin, Paul G.
中科院分区：
文献类型：
--
作者：
Kezic, Jelena Marie;Chen, Xiangting;McMenamin, Paul G.
目的.糖尿病视网膜病变（DR）是发达国家视力损害的主要原因。虽然DR被经典地描述为微血管疾病，但最近的证据表明视网膜小胶质细胞的变化是视网膜病变的早期特征。在我们的研究中，我们评估了非增殖性DR小鼠模型体内和体外小胶质细胞分布和形态的变化，并进一步研究了年龄和功能性趋化因子受体Cx（3）cr1缺失对这些变化进展的影响。为了分离三个变量的影响：糖尿病状态、年龄和Cx3cr1的作用，将Ins2（秋田）小鼠与Cx（3）cr1-eGFP报告小鼠杂交。在10、20、30和46周龄时对眼睛进行临床体内评估，并使用整体免疫荧光染色和共聚焦显微镜检查离体视网膜结构和GFP（+）小胶质细胞的排列。眼底、脉管系统或GFP（+）小胶质细胞分布的临床检查未发现任何与糖尿病状态相关的肉眼变化：然而，离体显微镜分析显示，Ins2（秋田）小鼠从10周龄开始，小胶质细胞网络组织发生改变，细胞形状变化的证据被经典地视为激活的迹象。这些变化在小胶质细胞缺乏Cx（3）cr1的老年糖尿病小鼠（Ins2（秋田）Cx（3）cr1（gfp/gfp）小鼠）中加剧。糖尿病状态和Cx（3）cr1缺陷导致Iba-1（+）透明细胞（玻璃体巨噬细胞）和视网膜下巨噬细胞聚集。这些数据表明，在没有明显视网膜病变和炎症的情况下，鼠视网膜小胶质细胞的变化响应于全身性糖尿病状态而发生。这些变化在缺乏Cx（3）cr1的小鼠中被夸大，表明fractalkine-Cx（3）cr1相互作用可能在增殖前DR的早期神经元变化中起作用（Invest Ophthalmol维斯Sci. 2013; 54：854 - 863）DOI：10.1167/iovs.12 - 10876
PURPOSE. Diabetic retinopathy (DR) is a major cause of visual impairment in developed countries. While DR has been described classically as a microvascular disease, recent evidence suggests that changes to retinal microglia are an early feature of retinopathy. In our study, we assessed changes in microglial distribution and morphology in vivo and ex vivo in a mouse model of non-proliferative DR, and further examined effects of age and the absence of the functional chemokine receptor Cx(3)cr1 on the progression of these changes.METHODS. To isolate the effects of the three variables: diabetic status, age, and role of Cx3cr1, the Ins2(Akita) mouse was crossed with Cx(3)cr1-eGFP reporter mice. Eyes were assessed clinically in vivo at 10, 20, 30, and 46 weeks of age, and the retinal structure and arrangement of GFP(+) microglia was examined ex vivo using whole mount immunofluorescence staining and confocal microscopy.RESULTS. Clinical examination of the fundus, vasculature, or GFP(+) microglial distribution did not reveal any macroscopic changes related to diabetic status: however, ex vivo microscopic analysis revealed alterations in microglial network organization, and evidence of cell shape changes regarded classically as signs of activation, in Ins2(Akita) mice from 10 weeks of age. These changes were exacerbated in older diabetic mice whose microglia lacked Cx(3)cr1 (Ins2(Akita) Cx(3)cr1(gfp/gfp) mice). Diabetic status and Cx(3)cr1 deficiency led to accumulations of Iba-1(+) hyalocytes (vitreal macrophages) and subretinal macrophages.CONCLUSIONS. These data showed that changes to murine retinal microglia occur in response to systemic diabetic status in the absence of overt retinopathy and inflammation. These changes are exaggerated in mice lacking Cx(3)cr1, suggesting fractalkine-Cx(3)cr1 interactions may have a role in early neuronal changes in preproliferative DR. (Invest Ophthalmol Vis Sci. 2013;54:854-863) DOI:10.1167/iovs.12-10876