Kitani K, Tanaka S, Zs-Nagy I
National Institute for Longevity Sciences, 36-3 Gengo, Morioka-cho, Obu-shi, Aichi 474, Japan.
Arch Gerontol Geriatr. 1998 May-Jun;26(3):257-73. doi: 10.1016/s0167-4943(98)00008-9.
The lateral diffusion coefficients of lipids (D(l)) and proteins (D(p)) were measured in hepatocyte plasma membranes in freshly prepared liver smears of Brown Norway (BN/BiRijHsd) rats by means of the fluorescence recovery after photobleaching (FRAP) method. D(l) was measured after staining with N-4-nitrobenzo-2-oxa-1,3-diazolyl phosphatidylethanolamine (NBD-PE) in 14 males in four age groups (4.3-30.4 months), and 12 females (5.0-31.0 months). D(p) was measured after the development of peroxide-induced autofluorescence (PIAF) in 16 males in four age groups (4.7-30.8 months) and 15 females in four age groups (4.0-28.1 months). Highly significant negative, linear age-correlations of D(l) (R=-0.972 in males, and -0.840 in females, respectively) were found, the average data being 1.39x10(-9) cm(2)/s in the young male rats, and only 9.26x10(-10) cm(2)/s in the oldest ones, whereas in females, these two extremes were 1.44x10(-9) and 8.97x10(-10) cm(2)/s, respectively. Protein lateral mobility also declined linearly with age; in the male rats it decreased from 2.7x10(-10) to 1.8.x10(-10) cm(2)/s (R=-0.903), while in the females it decreased from 2.64x10(-10) to 2.21x10(-10) cm(2)/s (R=-0.955) between the youngest and oldest age groups. There were also some characteristic differences in the fractional recoveries (FR%), too. These trends are comparable with those of F344 rats studied previously, supporting the conclusion that the changes in the lateral mobility of plasma membrane proteins and lipids represent important biomarkers of cellular aging also in the BN/BiRijHsd rats.
采用光漂白后荧光恢复（FRAP）法，在新鲜制备的棕色挪威（BN/BiRijHsd）大鼠肝脏涂片的肝细胞质膜中测量脂质（D(l)）和蛋白质（D(p)）的侧向扩散系数。在四个年龄组（4.3 - 30.4个月）的14只雄性和12只雌性（5.0 - 31.0个月）大鼠中，用N - 4 - 硝基苯 - 2 - 恶唑 - 1,3 - 二氮杂萘基磷脂酰乙醇胺（NBD - PE）染色后测量D(l)。在四个年龄组（4.7 - 30.8个月）的16只雄性和四个年龄组（4.0 - 28.1个月）的15只雌性大鼠中，在过氧化物诱导的自发荧光（PIAF）形成后测量D(p)。发现D(l)与年龄呈高度显著的负线性相关（雄性R = - 0.972，雌性R = - 0.840），年轻雄性大鼠的平均数据为1.39×10⁻⁹ cm²/s，而最老的大鼠仅为9.26×10⁻¹⁰ cm²/s，而在雌性中，这两个极端值分别为1.44×10⁻⁹和8.97×10⁻¹⁰ cm²/s。蛋白质侧向迁移率也随年龄呈线性下降；在雄性大鼠中，它从2.7×10⁻¹⁰降至1.8×10⁻¹⁰ cm²/s（R = - 0.903），而在雌性中，在最年轻和最老年龄组之间，它从2.64×10⁻¹⁰降至2.21×10⁻¹⁰ cm²/s（R = - 0.955）。在分数恢复率（FR%）方面也存在一些特征差异。这些趋势与先前研究的F344大鼠的趋势相似，支持了质膜蛋白质和脂质侧向迁移率的变化也是BN/BiRijHsd大鼠细胞衰老重要生物标志物的结论。
