Eye Care Centre, Leighton Hospital, Crewe, Cheshire, UK.
Br J Ophthalmol. 2010 Oct;94(10):1388-92. doi: 10.1136/bjo.2009.176032. Epub 2010 Aug 7.
A study was undertaken to determine and compare the F-actin staining patterns in the cells of the lamina cribrosa (LC) of normal, dexamethasone (DEX)-treated and glaucomatous dissected tissue and cell cultures.
About 30 dissected donor eyes and nine cell lines provided the human specimens; 25 eyes and 20 primary cell cultures provided the bovine material. Appropriate samples were exposed to 1×10⁻⁷ M DEX for up to 14 days. LC tissue and cells were stained with Phalloidin-Alexa 488 to identify F-actin, and all samples were examined by confocal or epifluorescent microscopy.
Both in the LC tissue and LC cell cultures the dominant actin arrangement was bundles of stress fibres. However, cross-linked actin networks (CLANs) were identified in the tissue and in culture. These were markedly increased by steroid treatment and were particularly large and abundant in cultures from glaucoma donors. CLANs were not associated with optic nerve head astrocytes.
The presence of abundant stress fibres in situ and in vitro highlights the biomechanical contribution of LC cells. However, the identification of CLANs in these cells shows that they are not exclusive to the trabecular meshwork, the only other place they have been found, and may have a role in glaucoma pathology.
本研究旨在确定并比较正常、地塞米松（DEX）处理和青光眼解剖组织及细胞培养物中，栅状带细胞（LC）中的 F-肌动蛋白染色模式。
约 30 个解剖供体眼和 9 个细胞系提供了人标本；25 只眼和 20 个原代细胞培养物提供了牛材料。将适当的样本暴露于 1×10⁻⁷ M 的 DEX 中，最长可达 14 天。用 Phalloidin-Alexa 488 对 LC 组织和细胞进行染色以鉴定 F-肌动蛋白，并用共聚焦或落射荧光显微镜检查所有样本。
在 LC 组织和 LC 细胞培养物中，优势肌动蛋白排列均为应力纤维束。然而，在组织和培养物中均鉴定出交联肌动蛋白网络（CLANs）。类固醇处理后，其数量明显增加，并且在来自青光眼供体的培养物中特别大且丰富。CLANs 与视神经头星形胶质细胞无关。
原位和体外存在丰富的应力纤维强调了 LC 细胞的生物力学贡献。然而，在这些细胞中鉴定出 CLANs 表明，它们不仅存在于小梁网（迄今为止唯一发现它们的地方）中，而且可能在青光眼病理中具有作用。
