Centre for Eye Research Australia, Department of Ophthalmology, University of Melbourne, Royal Victorian Eye and Ear Hospital, East Melbourne, Victoria, Australia.
Invest Ophthalmol Vis Sci. 2013 Apr 30;54(4):3028-36. doi: 10.1167/iovs.13-11865.
To assess the retinal macrophage response to cannulation of the anterior chamber (AC) and acute elevation of intraocular pressure (IOP) in adult mice.
Eyes from 12-month-old C57BL/6J WT mice were subject to IOP increase (50 mm Hg for 30 minutes) by direct cannulation of the AC. Fellow eyes were either cannulated without pressure increase or left untreated. Electroretinography was carried out prior to IOP elevation and 1 week later. Immunofluorescence staining was performed on frozen sections and retinal wholemounts 1 week after sham and IOP elevation. Eyes were assessed by epifluorescence and confocal microscopy and the density of vitreal hyalocytes and subretinal macrophages was calculated.
The density of hyalocytes and subretinal macrophages was significantly increased 1 week after IOP elevation and AC cannulation compared with naïve eyes. CD68 and MHC Class II expression was upregulated in both cannulated eyes and eyes with elevated IOP. Electroretinographic signals derived from retinal ganglion cells were significantly reduced in response to acute IOP elevation, but not in response to cannulation alone.
Cannulation of the AC causes an increase in hyalocyte density, microglial activation, and accumulation of macrophages in the subretinal space. These macrophage changes are similar to those observed in eyes subject to IOP elevation. Additional IOP elevation led to significant Müller cell activation, which was not evident after cannulation alone. These data highlight the importance of using appropriate controls in models of acute retinal injury.
评估成年小鼠前房(AC)插管和眼内压(IOP)急性升高对视网膜巨噬细胞的反应。
12 月龄 C57BL/6J WT 小鼠的眼睛通过直接 AC 插管使 IOP 升高(30 分钟内升高 50mmHg)。对侧眼睛要么不升高压力进行插管,要么不进行处理。在 IOP 升高前和 1 周后进行视网膜电图检查。在假手术和 IOP 升高后 1 周,对冷冻切片和视网膜全层进行免疫荧光染色。通过相差显微镜和共聚焦显微镜评估眼睛,并计算玻璃体内纤维细胞和视网膜下巨噬细胞的密度。
与正常眼相比,IOP 升高和 AC 插管后 1 周,玻璃体内纤维细胞和视网膜下巨噬细胞的密度显著增加。CD68 和 MHC Class II 在插管眼和 IOP 升高的眼中表达上调。急性 IOP 升高会导致视网膜神经节细胞的视网膜电图信号显著降低,但单独插管不会。
AC 插管会导致玻璃体内纤维细胞密度增加、小胶质细胞激活和视网膜下空间巨噬细胞积累。这些巨噬细胞变化与IOP 升高引起的变化相似。进一步的 IOP 升高会导致显著的 Müller 细胞激活,而单独插管后则不会出现这种情况。这些数据强调了在急性视网膜损伤模型中使用适当对照的重要性。