Raguz Marija, Mainali Laxman, O'Brien William J, Subczynski Witold K
Department of Biophysics, Medical College of Wisconsin, Milwaukee, WI 53226, USA; Department of Medical Physics and Biophysics, School of Medicine, University of Split, Split, Croatia.
Department of Biophysics, Medical College of Wisconsin, Milwaukee, WI 53226, USA.
Exp Eye Res. 2015 Mar;132:78-90. doi: 10.1016/j.exer.2015.01.018. Epub 2015 Jan 21.
The results reported here clearly document changes in the properties and the organization of fiber-cell membrane lipids that occur with age, based on electron paramagnetic resonance (EPR) analysis of lens membranes of clear lenses from donors of age groups from 0 to 20, 21 to 40, and 61 to 80 years. The physical properties, including profiles of the alkyl chain order, fluidity, hydrophobicity, and oxygen transport parameter, were investigated using EPR spin-labeling methods, which also provide an opportunity to discriminate coexisting lipid domains and to evaluate the relative amounts of lipids in these domains. Fiber-cell membranes were found to contain three distinct lipid environments: bulk lipid domain, which appears minimally affected by membrane proteins, and two domains that appear due to the presence of membrane proteins, namely boundary and trapped lipid domains. In nuclear membranes the amount of boundary and trapped phospholipids as well as the amount of cholesterol in trapped lipid domains increased with the donors' age and was greater than that in cortical membranes. The difference between the amounts of lipids in domains uniquely formed due to the presence of membrane proteins in nuclear and cortical membranes increased with the donors' age. It was also shown that cholesterol was to a large degree excluded from trapped lipid domains in cortical membranes. It is evident that the rigidity of nuclear membranes was greater than that of cortical membranes for all age groups. The amount of lipids in domains of low oxygen permeability, mainly in trapped lipid domains, were greater in nuclear than cortical membranes and increased with the age of donors. These results indicate that the nuclear fiber cell plasma membranes were less permeable to oxygen than cortical membranes and become less permeable to oxygen with age. In clear lenses, age-related changes in the lens lipid and protein composition and organization appear to occur in ways that increase fiber cell plasma membrane resistance to oxygen permeation.
基于对年龄在0至20岁、21至40岁以及61至80岁捐赠者的透明晶状体膜进行电子顺磁共振(EPR)分析,本文报道的结果清楚地证明了随着年龄增长,纤维细胞膜脂质的性质和组织发生了变化。使用EPR自旋标记方法研究了包括烷基链顺序、流动性、疏水性和氧传输参数等物理性质,该方法还提供了区分共存脂质结构域以及评估这些结构域中脂质相对含量的机会。发现纤维细胞膜包含三种不同的脂质环境:大体脂质结构域,似乎受膜蛋白影响最小;以及由于膜蛋白的存在而出现的两个结构域,即边界脂质结构域和捕获脂质结构域。在核膜中,边界磷脂和捕获磷脂的量以及捕获脂质结构域中胆固醇的量随捐赠者年龄增加而增加,且大于皮质膜中的量。核膜和皮质膜中由于膜蛋白的存在而独特形成的结构域中脂质含量的差异随捐赠者年龄增加而增大。还表明,皮质膜中捕获脂质结构域很大程度上排除了胆固醇。显然,所有年龄组的核膜刚性均大于皮质膜。低氧渗透性结构域中的脂质量,主要是捕获脂质结构域中的脂质,在核膜中比皮质膜中更多,并且随捐赠者年龄增加而增加。这些结果表明,核纤维细胞质膜对氧的渗透性低于皮质膜,并且随着年龄增长对氧的渗透性降低。在透明晶状体中,晶状体脂质和蛋白质组成及组织的年龄相关变化似乎以增加纤维细胞质膜对氧渗透的抵抗力的方式发生。