Muller Cell Expression of Genes Implicated in Proliferative Vitreoretinopathy Is Influenced by Substrate Elastic Modulus
Muller Cell Expression of Genes Implicated in Proliferative Vitreoretinopathy Is Influenced by Substrate Elastic Modulus
复制标题
DOI：
10.1167/iovs.11-8450
复制
发表时间：
2012-05-01
影响因子：
4.4
通讯作者：
Foster, William J.
中科院分区：
文献类型：
--
作者：
Davis, Joshua T.;Wen, Qi;Foster, William J.
目的.基质硬度越来越被认为是细胞和组织功能的重要因素。为了更好地了解Muller细胞的机械敏感性及其与玻璃体视网膜疾病的关系，我们研究了在不同弹性模量的基质上培养的Muller细胞的形态、繁殖和基因表达。将条件永生化小鼠Muller细胞系在具有校准杨氏模量的层粘连蛋白包被的聚丙烯酰胺基质上培养。玻璃被用作对照。相差，荧光和原子力显微镜被用来研究细胞形态和繁殖。细胞外基质（ECM）基因的表达进行了分析，采用定量逆转录PCR。基质刚度对Muller细胞的粘附面积、刚度和增殖均有影响，但影响程度不同，敏感性范围也不同。在所测试的85个ECM基因中，11个显示mRNA表达的连续> 4倍增加或减少，其作为基底弹性模量的函数。四个基因的变化具有统计学意义：结缔组织生长因子（Ctgf，P = 0.04）、腱生蛋白C（Tnc，P = 0.035）、胶原蛋白I α 1（Col 1a 1，P = 0.0001）和胶原蛋白IV α 3（Col 4a 3，P = 0.05），所有基因在较软的基质上的表达均增加。Muller细胞的形态、细胞骨架完整性和基因调节随着基质硬度的变化而发生显着变化。局部组织弹性模量的变化可能在玻璃体视网膜疾病中起作用。这些发现也可能对改善视网膜修复体整合的策略以及干细胞疗法，特别是针对转录调节因子雅普和TAZ的干细胞疗法具有影响。（Invest Ophthalmol维斯科学。2012;53：3014-3019）DOI：10.1167/iovs.11-8450
PURPOSE. Matrix stiffness is recognized increasingly as a significant factor in cell and tissue function. To understand better the mechanosensitivity of Muller cells and its association with vitreoretinal disorders, we examined morphology, propagation, and expression of genes in Muller cells that were cultured on substrates of varying elastic moduli.METHODS. A conditionally immortalized mouse Muller cell line was cultured on laminin-coated polyacrylamide substrates with calibrated Young's moduli. Glass was used as a control. Phase contrast, fluorescence, and atomic force microscopy were used to study cell morphology and propagation. Expression of extracellular matrix (ECM) genes was analyzed using quantitative reverse-transcription PCR.RESULTS. The adherent area, stiffness, and propagation of Muller cells all are affected by matrix stiffness, but to different extents and with different ranges of sensitivity. Of 85 ECM genes tested 11 showed a continuous > 4-fold increase or decrease in mRNA expression as a function of the substrate elastic modulus. The changes were statistically significant in four genes: connective tissue growth factor (Ctgf, P = 0.04), tenascin C (Tnc, P = 0.035), Collagen I alpha 1 (Col1a1, P = 0.0001), and Collagen IV alpha 3 (Col4a3, P = 0.05), with all showing increased expression on softer substrates.CONCLUSIONS. There are significant changes in morphology, cytoskeletal integrity, and gene regulation in Muller cells as a function of the stiffness of the substrate. Changes in local tissue elastic modulus may have a role in vitreoretinal disorders. These findings also may have implications for strategies for improved integration of retinal prosthetics, and for stem cell therapies, particularly targeting the transcriptional regulators YAP and TAZ. (Invest Ophthalmol Vis Sci. 2012;53:3014-3019) DOI:10.1167/iovs.11-8450