Yechiel E, Barenholz Y, Henis Y I
J Biol Chem. 1985 Aug 5;260(16):9132-6.
The effects of aging and of liposome treatment on the lateral mobility of phospholipids and proteins in the plasma membrane of cultured rat heart myocytes were studied by fluorescence photobleaching recovery. Both the mobile fraction (R) and the lateral diffusion coefficient (D) of the fluorescent phospholipid N-4-nitrobenzo-2-oxa-1,3-diazolyl phosphatidylethanolamine were found to depend on the culture's age. Aged myocyte cultures (15 days old) demonstrated higher R and lower D as compared with young ones (5 days old). Treatment of aged cultures with phosphatidylcholine (PC) liposomes, which increases the PC/sphingomyelin (SM) ratio and decreases the cholesterol level, reversed the D value to the level observed in young cultures and decreased R below the value encountered in young cells. Treatments with SM liposomes (which induce cholesterol depletion without altering the PC/SM ratio) and with PC/cholesterol (1:0.9) liposomes (which increase the PC/SM ratio without cholesterol depletion) have indicated that the PC-liposome effect is due to changes in both the PC/SM ratio and in the cholesterol level. Analogous experiments on the mobility of succinyl-concanavalin A receptors yielded similar effects on R, without altering the D value. The changes in the D and R values of the markers studied are most likely initiated by the observed alterations in the myocyte lipid composition under the conditions employed. The possible involvement of changes in the organization of membrane lipids in domains in the observed phenomena is discussed.
通过荧光光漂白恢复技术研究了衰老和脂质体处理对培养的大鼠心肌细胞质膜中磷脂和蛋白质侧向流动性的影响。发现荧光磷脂N-4-硝基苯-2-恶唑-1,3-二氮杂萘基磷脂酰乙醇胺的可移动部分(R)和侧向扩散系数(D)均取决于培养物的年龄。与年轻的培养物(5天大)相比,衰老的心肌细胞培养物(15天大)表现出更高的R和更低的D。用磷脂酰胆碱(PC)脂质体处理衰老培养物,可增加PC/鞘磷脂(SM)的比例并降低胆固醇水平,使D值恢复到年轻培养物中观察到的水平,并使R降低到低于年轻细胞中的值。用SM脂质体(诱导胆固醇消耗而不改变PC/SM比例)和PC/胆固醇(1:0.9)脂质体(增加PC/SM比例而不消耗胆固醇)进行的处理表明,PC脂质体效应是由于PC/SM比例和胆固醇水平的变化所致。对琥珀酰伴刀豆球蛋白A受体流动性的类似实验对R产生了类似的影响,而未改变D值。在所采用的条件下,研究的标记物的D和R值的变化很可能是由观察到的心肌细胞脂质组成的改变引发的。讨论了膜脂质组织变化在观察到的现象中的可能作用。