Department of Ophthalmology, University of Basel, Switzerland.
Proteomics Core Facility, Biozentrum, University of Basel, Switzerland.
Exp Eye Res. 2020 Dec;201:108326. doi: 10.1016/j.exer.2020.108326. Epub 2020 Nov 2.
The Descemet's membrane (DM) and the lens capsule (LC) are two ocular basement membranes (BMs) that are essential in maintaining stability and structure of the cornea and lens. In this study, we investigated the proteomes and biomechanical properties of these two materials to uncover common and unique properties. We also screened for possible protein changes during diabetes. LC-MS/MS was used to determine the proteomes of both BMs. Biomechanical measurements were conducted by atomic force microscopy (AFM) in force spectroscopy mode, and complemented with immunofluorescence microscopy. Proteome analysis showed that all six existing collagen IV chains represent 70% of all LC-protein, and are thus the dominant components of the LC. The DM on the other hand is predominantly composed of a single protein, TGF-induced protein, which accounted for around 50% of all DM-protein. Four collagen IV-family members in DM accounted for only 10% of the DM protein. Unlike the retinal vascular BMs, the LC and DM do not undergo significant changes in their protein compositions during diabetes. Nanomechanical measurements showed that the endothelial/epithelial sides of both BMs are stiffer than their respective stromal/anterior-chamber sides, and both endothelial and stromal sides of the DM were stiffer than the epithelial and anterior-chamber sides of the LC. Long-term diabetes did not change the stiffness of the DM and LC. In summary, our analyses show that the protein composition and biomechanical properties of the DM and LC are different, i.e., the LC is softer than DM despite a significantly higher concentration of collagen IV family members. This finding is unexpected, as collagen IV members are presumed to be responsible for BM stiffness. Diabetes had no significant effect on the protein composition and the biomechanical properties of both the DM and LC.
Descemet 膜(DM)和晶状体囊(LC)是两种眼基底膜(BMs),对于维持角膜和晶状体的稳定性和结构至关重要。在本研究中,我们研究了这两种材料的蛋白质组学和生物力学特性,以揭示其共同和独特的特性。我们还筛选了糖尿病过程中可能发生的蛋白质变化。LC-MS/MS 用于确定这两种 BM 的蛋白质组。原子力显微镜(AFM)在力谱模式下进行生物力学测量,并辅以免疫荧光显微镜。蛋白质组分析表明,所有 6 种现有的 IV 型胶原链代表了 LC 中所有蛋白质的 70%,因此是 LC 的主要成分。另一方面,DM 主要由一种蛋白质,即 TGF 诱导蛋白组成,占所有 DM 蛋白的 50%左右。DM 中的四个 IV 型胶原家族成员仅占 DM 蛋白的 10%。与视网膜血管 BM 不同,LC 和 DM 在糖尿病过程中其蛋白质组成没有发生显著变化。纳米力学测量表明,两种 BM 的内皮/上皮侧均比各自的基质/前房侧更硬,DM 的内皮和基质侧均比 LC 的上皮和前房侧更硬。长期糖尿病并没有改变 DM 和 LC 的硬度。总之,我们的分析表明,DM 和 LC 的蛋白质组成和生物力学特性不同,即尽管 LC 中 IV 型胶原家族成员的浓度明显更高,但 LC 比 DM 更柔软。这一发现出人意料,因为 IV 型胶原成员被认为是 BM 硬度的主要原因。糖尿病对 DM 和 LC 的蛋白质组成和生物力学特性均无显著影响。