The University of Queensland, Perinatal Research Centre, Brisbane, Queensland, Australia.
Clin Exp Ophthalmol. 2011 Aug;39(6):555-63. doi: 10.1111/j.1442-9071.2010.02488.x. Epub 2011 Feb 23.
To investigate retinal cell population changes under chronic elevated intraocular pressure in an inducible mouse model of glaucoma.
Chronic unilateral ocular hypertension was induced in 40 C57BL6/J mice by ablation of the limbal episcleral veins. After 5, 20, 40 and 60 days of elevated intraocular pressure, specific retinal cell types were identified and/or quantified by immunohistochemistry for protein kinase C α, glial fibrillary acidic protein, parvalbumin and calretinin. Apoptotic cells were identified by TUNEL and cleaved caspase-3 immunohistochemistry.
Elevations in intraocular pressure in the range 22-30 mmHg were developed and sustained in mice for up to 60 days. Protein kinase C α immunoreactivity localized to bipolar cells was unchanged. We observed a rapid increase in glial fibrillary acidic protein expression in Müller cells and a progressive loss of parvalbumin-labelled ganglion cells. After 60 days of elevated intraocular pressure, calretinin-immunoreactive cell counts declined by 55.4% and 46.4% in the inner nuclear and ganglion cell layers, respectively. However, at all time points examined, the markers of cell death were only observed in the ganglion cell layer, not in the inner nuclear layer.
In addition to ganglion cell death and reactive Müller cell changes, chronic experimental elevation of intraocular pressure alters calcium-binding protein immunohistochemistry in amacrine cells. However, these changes are not indicative of amacrine cell loss but may represent early indicators of cellular distress that precede physiological dysfunction or cell death.
在青光眼诱导型小鼠模型中,研究慢性高眼压下视网膜细胞群体的变化。
通过切除角膜缘巩膜静脉,在 40 只 C57BL6/J 小鼠中诱导慢性单侧眼高压。在眼压升高 5、20、40 和 60 天后,通过免疫组织化学法对蛋白激酶 Cα、胶质纤维酸性蛋白、副甲状腺蛋白和钙调蛋白鉴定和/或定量特定的视网膜细胞类型。通过 TUNEL 和裂解半胱氨酸天冬氨酸蛋白酶-3 免疫组织化学法鉴定凋亡细胞。
在高达 60 天的时间内,小鼠眼内压升高至 22-30mmHg,并持续升高。双极细胞定位的蛋白激酶 Cα免疫反应性保持不变。我们观察到 Müller 细胞中胶质纤维酸性蛋白表达迅速增加,副甲状腺蛋白标记的节细胞逐渐丧失。在眼压升高 60 天后,内核层和节细胞层中 calretinin 免疫反应性细胞计数分别下降了 55.4%和 46.4%。然而,在所有检查的时间点,细胞死亡的标志物仅在节细胞层中观察到,而在内核层中未观察到。
除了节细胞死亡和反应性 Müller 细胞变化外,慢性实验性眼压升高还改变了无长突细胞中钙结合蛋白的免疫组织化学。然而,这些变化并不表明无长突细胞丢失,而是可能代表细胞功能障碍或细胞死亡之前的早期细胞应激的指标。