Retina Service, Angiogenesis Laboratory, Massachusetts Eye and Ear Infirmary, Department of Ophthalmology, Harvard Medical School, Boston, Massachusetts, United States.
Invest Ophthalmol Vis Sci. 2014 May 22;55(7):4165-74. doi: 10.1167/iovs.14-14238.
To evaluate the potential for mouse genetic background to effect photoreceptor cell death in response to experimental retinal detachment (RD).
Retinal detachment was induced in three inbred mouse strains (C57BL/6, BALB/c, and B6129SF2) by subretinal injection of sodium hyaluronate. A time course of photoreceptor cell death was assessed by TUNEL assay. Total photoreceptor cell death was analyzed through comparing the outer nuclear layer (ONL)/inner nuclear layer (INL) ratio 7 days post RD. Western blot analysis or quantitative real-time PCR (qPCR) were performed to assess cell death signaling, expression of endogenous neurotrophin, and levels of apoptosis inhibitors 24 hours after RD. Inflammatory cytokine secretion and inflammatory cell infiltration were quantified by ELISA and immunostaining, respectively.
The peak of photoreceptor cell death after RD was at 24 hours in all strains. Photoreceptor cell death as well as monocyte chemoattractant protein 1 and interleukin 6 secretion at 24 hours after RD was the highest in BALB/c, followed in order of magnitude by C57BL/6 and B6129SF2. Conversely, nerve growth factor expression and ONL/INL ratio were the lowest in BALB/c. Apoptosis signaling was higher in C57BL/6, whereas necroptosis signaling was higher in C57BL/6 and BALB/c. Autophagic signaling was higher in BALB/c. X-linked inhibitor of apoptosis (XIAP) and survivin protein levels were lower in C57BL/6 and BALB/c, respectively. Macrophage/microglia infiltration was higher in C57BL/6 and BALB/c at 24 hours after RD.
Photoreceptor cell death after RD was significantly different among the three strains, suggesting the presence of genetic factors that affect photoreceptor cell death after RD.
评估小鼠遗传背景对实验性视网膜脱离（RD）后光感受器细胞死亡的潜在影响。
通过视网膜下注射透明质酸钠在三种近交系小鼠（C57BL/6、BALB/c 和 B6129SF2）中诱导视网膜脱离。通过 TUNEL 测定法评估光感受器细胞死亡的时间过程。通过比较 RD 后 7 天的外核层（ONL）/内核层（INL）比来分析总光感受器细胞死亡。Western blot 分析或定量实时 PCR（qPCR）用于评估 RD 后 24 小时的细胞死亡信号、内源性神经营养因子的表达和凋亡抑制剂的水平。通过 ELISA 和免疫染色分别定量炎性细胞因子分泌和炎性细胞浸润。
所有品系在 RD 后 24 小时达到光感受器细胞死亡的高峰。RD 后 24 小时的光感受器细胞死亡以及单核细胞趋化蛋白 1 和白细胞介素 6 的分泌在 BALB/c 中最高，其次是 C57BL/6 和 B6129SF2。相反，BALB/c 中的神经生长因子表达和 ONL/INL 比最低。C57BL/6 中的凋亡信号较高，而 C57BL/6 和 BALB/c 中的坏死信号较高。BALB/c 中的自噬信号较高。C57BL/6 和 BALB/c 中的 X 连锁凋亡抑制剂（XIAP）和生存素蛋白水平较低。RD 后 24 小时的巨噬细胞/小胶质细胞浸润在 C57BL/6 和 BALB/c 中较高。
三种品系之间 RD 后光感受器细胞死亡有显著差异，表明存在影响 RD 后光感受器细胞死亡的遗传因素。
